Artificial secretory peptides for heterologous protein production

Abstract

Provided herein are pre-protein signal peptides that direct the secretion of expressed payload proteins from Lactobacillus bacteria. The pre-protein signal peptides described herein are designed to increase payload secretion over known signal peptides while minimizing or reducing the impact on the fitness of the bacterial strain. Also provided herein are methods for using the pre-protein signal peptides provided herein.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 63 / 477,959, filed December 30, 2022, which is incorporated herein by reference in its entirety.

[0002] Reference to an electronically submitted sequence listing This application contains a Sequence Listing that has been submitted electronically in XML format, and is incorporated herein by reference in its entirety. The XML copy, created on December 29, 2023, is named "TNZ-008WO_SL.xml" and is 273,334 bytes in size.

[0003] FIELD OF THE INVENTION Embodiments provided herein relate generally to signal peptides, and more specifically to synthetic preprotein signal peptides that increase secretion of recombinant proteins in Lactobacillus. [Background technology]

[0004] Bacteria are routinely used as hosts for producing proteins for research, therapeutic, and industrial purposes. The first step in the secretion of a desired target protein into the growth medium is its transport across the cytoplasmic membrane. In bacteria, two major transport pathways are utilized for protein transport across the plasma membrane: the general secretory pathway, or Sec pathway, and the twin-arginine translocation, or Tat pathway. Routing a protein to one of these alternative protein transport systems requires the fusion of a Sec- or Tat-specific signal peptide to the amino terminus of the desired target protein. Signal peptides are required for targeting and membrane translocation by the respective protein translocases, but they also affect the biosynthesis, folding rate, and stability of the respective payload protein. Currently, it is impossible to predict which signal peptide will function best for a given payload protein and a given bacterial expression host.

[0005] Furthermore, while increased secretory production of payload proteins through the use of signal peptides is desirable, there are additional factors that must be considered when developing methods for producing a particular payload protein. For example, signal peptides that result in increased secretion of a recombinant payload protein may be harmful to the host bacterium. Therefore, an effective level of increased protein secretion that will maintain the overall health, or "fitness," of the host bacterium must be identified. Embodiments of the present disclosure address these needs and more. In particular, the signal peptides of the present disclosure have been optimized to increase the yield of recombinant payload proteins over known signal peptides while minimizing the impact on the fitness of the host bacterium. Summary of the Invention

[0006] In some embodiments, a pre-protein signal peptide is provided. In some embodiments, the pre-protein signal peptide comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence selected from SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:148, or SEQ ID NO:180. In some embodiments, the pre-protein signal peptide comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:6. In some embodiments, the pre-protein signal peptide comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:7. In some embodiments, the pre-protein signal peptide comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:14. In some embodiments, the preprotein signal peptide comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 17. In some embodiments, the preprotein signal peptide comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 18. In some embodiments, the preprotein signal peptide comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 19. In some embodiments, the preprotein signal peptide comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 20. In some embodiments, the preprotein signal peptide comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 47.In some embodiments, the preprotein signal peptide comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 48. In some embodiments, the preprotein signal peptide comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 148. In some embodiments, the preprotein signal peptide comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 180.

[0007] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide and Z1 is a payload protein. In some embodiments, X1 comprises an amino acid sequence having at least 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:148, or SEQ ID NO:180. In some embodiments, Z1 is selected from the group consisting of an antimicrobial peptide, catestatin, CRAMP, hBD1, hBD3, β-catethritin, dermaseptin, IL-10, IL-22, ClbS, TIGIT, TFF1, TFF2, TFF3, an anti-CCL2 antibody, an anti-CCL2 antibody fragment, an anti-IL23 antibody, an anti-IL23 antibody fragment, an anti-TNFα antibody, an anti-TNFα antibody fragment, GLP2, IGF1, leptin, and casein.

[0008] In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:6 and Z1 comprises the amino acid sequence of SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, SEQ ID NO:120, or a fragment thereof.

[0009] In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7 and Z1 comprises the amino acid sequence of SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, SEQ ID NO:120, or a fragment thereof.

[0010] In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14 and Z1 comprises the amino acid sequence of SEQ ID NO: 21, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, or a fragment thereof.

[0011] In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:6 and Z1 comprises the amino acid sequence of SEQ ID NO:25.

[0012] In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7 and Z1 comprises the amino acid sequence of SEQ ID NO:25.

[0013] In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:14 and Z1 comprises the amino acid sequence of SEQ ID NO:25.

[0014] In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:17 and Z1 comprises the amino acid sequence of SEQ ID NO:25.

[0015] In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:18 and Z1 comprises the amino acid sequence of SEQ ID NO:25.

[0016] In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:19 and Z1 comprises the amino acid sequence of SEQ ID NO:25.

[0017] In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:47 and Z1 comprises the amino acid sequence of SEQ ID NO:25.

[0018] In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:48 and Z1 comprises the amino acid sequence of SEQ ID NO:25.

[0019] In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 148 and Z1 comprises the amino acid sequence of SEQ ID NO: 25.

[0020] In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 180 and Z1 comprises the amino acid sequence of SEQ ID NO: 25.

[0021] In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:20 and Z1 comprises the amino acid sequence of SEQ ID NO:24, SEQ ID NO:283, SEQ ID NO:284, or SEQ ID NO:285.

[0022] In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7 and Z1 comprises the amino acid sequence of SEQ ID NO:22, SEQ ID NO:59, or SEQ ID NO:282.

[0023] In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14 and Z1 comprises the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282.

[0024] In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 148 and Z1 comprises the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282.

[0025] In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 180 and Z1 comprises the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282.

[0026] In some embodiments, a nucleic acid molecule is provided. In some embodiments, the nucleic acid molecule encodes a signal peptide provided herein.

[0027] In some embodiments, a nucleic acid molecule is provided. In some embodiments, the nucleic acid molecule encodes a polypeptide provided herein.

[0028] In some embodiments, a bacterium is provided. In some embodiments, the bacterium comprises a heterologous nucleic acid molecule encoding a polypeptide provided herein.

[0029] In some embodiments, a pharmaceutical composition is provided, hi some embodiments, the pharmaceutical composition comprises a bacterium provided herein and a pharmaceutically acceptable carrier.

[0030] In some embodiments, a method for producing a protein is provided, which method includes transfecting a bacterium with a nucleic acid molecule encoding a polypeptide provided herein to produce a bacterium containing the nucleic acid molecule, and culturing the bacterium containing the nucleic acid molecule under conditions sufficient to grow the bacterium, wherein the bacterium expresses and secretes the payload protein.

[0031] In some embodiments, a method of producing a protein in a subject is provided, in some embodiments, the method comprises administering to the subject a bacterium provided herein, or a pharmaceutical composition provided herein, wherein the bacterium is live, and wherein the live bacterium produces the protein in the subject.

[0032] In some embodiments, there is provided a method for treating a disease or disorder in a subject in need thereof, hi some embodiments, the method comprises administering to the subject a therapeutically effective amount of a bacterium described herein or a pharmaceutical composition described herein.

[0033] In some embodiments, there is provided a method of treating inflammation in a subject in need thereof, hi some embodiments, the method comprises administering to the subject a therapeutically effective amount of a bacterium described herein or a pharmaceutical composition described herein.

[0034] In some embodiments, there is provided a method for treating inflammation in a subject in need thereof. In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a bacterium comprising a heterologous nucleic acid molecule encoding a polypeptide having the formula X1-Z1, wherein X1 comprises the amino acid sequence of SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:14, or SEQ ID NO:20, and Z1 comprises the amino acid sequence of SEQ ID NO:21, SEQ ID NO:24, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120.

[0035] In some embodiments, there is provided a method for treating obesity in a subject in need thereof, hi some embodiments, the method comprises administering to the subject a therapeutically effective amount of a bacterium described herein or a pharmaceutical composition described herein.

[0036] In some embodiments, there is provided a method for treating obesity in a subject in need thereof. In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a bacterium comprising a heterologous nucleic acid molecule encoding a polypeptide having the formula X1-Z1, wherein X1 comprises the amino acid sequence of SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 47, or SEQ ID NO: 48, and Z1 comprises the amino acid sequence of SEQ ID NO: 25. [Brief explanation of the drawings]

[0037] [Figure 1] The effect of a panel of unique signal peptides (BP) on anti-TNFα antibody secretion and the doubling time of bacterial strains is shown compared to known signal peptides (A). The dotted line represents the doubling time of untransfected wild-type bacteria. Data points for peptide I and peptide O overlap. [Figure 2] The effects of signal peptides F (SEQ ID NO: 6), G (SEQ ID NO: 7), and N (SEQ ID NO: 14) on the secretion of various payload proteins and the doubling time of bacterial strains are shown in comparison to the known signal peptide A (SEQ ID NO: 1). [Figure 3] 1 shows the distribution of leptin secretion in the signal peptide library. [Figure 4]The effect of lead candidate signal peptides on leptin secretion and doubling time of bacterial strains is shown in comparison with the known signal peptide A (SEQ ID NO: 1). [Figure 5] The distribution of IL-10 secretion from the signal peptide library is shown. [Figure 6] The effect of SEQ ID NO: 20 on IL-10 secretion and doubling time of bacterial strains is shown in comparison to the known signal peptide SEQ ID NO: 1. A shows a comparison of IL-10 secretion. B shows a comparison of the change in doubling time compared to wild-type untransfected bacteria. [Figure 7] 1 shows the distribution of mouse IL-22 secretion from the signal peptide library. [Figure 8] The effect of signal peptides A2 (SEQ ID NO: 148) and D2 (SEQ ID NO: 180) on murine IL-22 secretion and the doubling time of the bacterial strain is shown compared to signal peptide G (SEQ ID NO: 7). The dotted line represents the doubling time of the WT bacterial strain. [Figure 9A] Figure 1 shows the effect of signal peptides A2 (SEQ ID NO: 148) and D2 (SEQ ID NO: 180) on the secretion of additional protein targets and the doubling time of bacterial strains. Human IL-22 secretion is shown compared to signal peptide G (SEQ ID NO: 7). [Figure 9B] Figure 1 shows the effect of signal peptides A2 (SEQ ID NO: 148) and D2 (SEQ ID NO: 180) on the secretion of additional protein targets and the doubling time of bacterial strains. Leptin secretion is shown in comparison to signal peptide lib-10 (SEQ ID NO: 19) and signal peptide A (SEQ ID NO: 1). DETAILED DESCRIPTION OF THE INVENTION

[0038] The present disclosure presents a solution to the above-mentioned problems by providing novel synthetic signal peptides that direct the secretion of proteins or peptides expressed in Bacillus bacteria. The disclosed signal peptides overcome the performance variability caused by previously characterized natural signal peptides and can be used to promote the production and secretion of any protein or peptide in bacteria.

[0039] The disclosed synthetic preprotein signal peptides increase the secretion of any recombinant protein in Lactobacillus bacteria. The use of synthetic preprotein signal peptides can further improve the secretion of payload proteins, for example, by promoting translocation across the cytoplasmic membrane. Advantageously, the signal peptides disclosed herein have been generated and optimized to promote the secretion of payload proteins from Lactobacillus bacteria. The use of the disclosed synthetic preprotein signal peptides can be used to achieve increased secretion of desired payloads into any bacterially compatible environment, such as therapeutics, agriculture, or food.

[0040] Before describing the compositions and methods of the present invention, it is to be understood that the scope of the present invention is not limited to the specific processes, compositions, or methodologies described herein, as they may vary. It is also to be understood that the terminology used in the description is merely for the purpose of describing particular versions or embodiments, and is not intended to limit the scope of the present invention. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the methods and systems disclosed herein, the preferred methods, devices, and materials are described below.

[0041] definition The following explanations of terms and methods are provided to better describe the present disclosure and to guide those of ordinary skill in the art in the practice of the present disclosure.

[0042] As used herein, "comprising" means "including," and the singular forms "a," "an," or "the" include plural references unless the context clearly dictates otherwise. For example, a reference to "comprising a therapeutic agent" includes one or more such therapeutic agents. The term "or" refers to a single element of the alternative elements referenced, unless the context clearly dictates otherwise. For example, the phrase "A or B" refers to A alone or B alone. The phrase "A, B, or a combination thereof" refers to A alone, B alone, or the combination of A and B. Similarly, "one or more of A and B" refers to A, B, or both A and B in combination. The phrase "A and B" refers to A and B in combination. Furthermore, the various elements, features, and steps discussed herein, as well as other known equivalents of each such element, feature, or step, can be mixed and matched by those skilled in the art to perform methods in accordance with the principles described herein. In particular embodiments, some of the various elements, features, and steps are specifically included, while others are specifically excluded.

[0043] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, suitable methods and materials are described below. The materials, methods, and examples are illustrative only and are not intended to be limiting. All references cited herein are incorporated by reference in their entirety.

[0044] It should be understood that in some instances, numerical values ​​expressing quantities of ingredients, properties such as molecular weight, reaction conditions, and the like, used to describe and claim particular embodiments are in some cases modified by the term "about" or "approximately." For example, "about" or "approximately" may indicate a variation of ±5% of the value it describes. Accordingly, in some embodiments, the numerical parameters set forth herein are approximations that may vary depending on the desired properties of the particular embodiment. Notwithstanding that the numerical ranges and parameters setting forth broad ranges of some examples are approximations, the numerical values ​​set forth in the specific examples are reported as precisely as practicable. Recitation of ranges of values ​​herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within that range.

[0045] In order to facilitate review of the various embodiments of the disclosure, the following explanations of specific terms are provided:

[0046] As used herein, "bacteria" (plural) and "bacteria" (singular) refer to unicellular prokaryotic microorganisms. Bacterial cells are generally surrounded by two protective coatings: an outer cell wall and an inner cell membrane. Bacteria may be classified according to Gram staining, which identifies bacteria by the composition of their cell wall. Gram-positive bacteria do not have an outer membrane, while Gram-negative bacteria do not. Bacteria generally replicate by binary fission. In binary fission, a parent cell makes copies of its DNA and doubles its cellular contents to grow larger. The cell then divides, extruding excess cellular contents and creating two daughter cells. Some bacteria utilize other processes, such as budding. In some embodiments, the bacterium is a wild-type natural isolate of bacteria. In some embodiments, the bacterium is an experimental bacterial strain that has undergone a breeding process of mutagenesis and selection. As used herein, "bacteria" refers to any known wild-type or laboratory bacterial strain.

[0047] As used herein, "Lactobacillus bacteria" refers to a genus of Gram-positive, aerotolerant anaerobes or microaerophilic, rod-shaped, non-spore-forming bacteria that constitute an important component of the human and animal microbiota. Examples of Lactobacillus bacteria include Lactobacillus acetotolerans, Lactobacillus acidifarinae, Lactobacillus acidipiscis, Lactobacillus acidophilus, Lactobacillus agilis, Lactobacillus algidus, Lactobacillus alimentarius, Lactobacillus alvei, Lactobacillus alvi, Lactobacillus amylolyticus, Lactobacillus amylophilus, Lactobacillus amylotrophicus, Lactobacillus amylovorus, Lactobacillus animalis, Lactobacillus animata, Lactobacillus antri, Lactobacillus apinorum, Lactobacillus apis, Lactobacillus apodemi, Lactobacillus aquaticus, Lactobacillus aviarius, Lactobacillus backii, Lactobacillus bifermentans, Lactobacillus bombi, Lactobacillus bombicola, Lactobacillus brantae, Lactobacillus brevis, Lactobacillus brevisimilis, Lactobacillus buchneri, Lactobacillus cacaonum, Lactobacillus camelliae, Lactobacillus capillatus, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus zeae, Lactobacillus catenefornis, Lactobacillus ceti, Lactobacilluscoleohominis, Lactobacillus colini, Lactobacillus collinoides, Lactobacillus composti, Lactobacillus concavus, Lactobacillus coryniformis, Lactobacillus crispatus, Lactobacillus crustorum, Lactobacillus curieae, Lactobacillus curvatus, Lactobacillus delbrueckii, Lactobacillus dextrinicus, Lactobacillus diolivorans, Lactobacillus equi, Lactobacillus equicursoris, Lactobacillus equigenerosi, Lactobacillus fabifermentans, Lactobacillus faecis, Lactobacillus faeni, Lactobacillus farciminis, Lactobacillus farraginis, Lactobacillus fermentum, Lactobacillus floricola, Lactobacillus florum, Lactobacillus formosensis, Lactobacillus fornicalis, Lactobacillus fructivorans, Lactobacillus frumenti, Lactobacillus fuchuensis, Lactobacillus furfuricola, Lactobacillus futsaii, Lactobacillus gallinarum, Lactobacillus gasseri, Lactobacillus gastricus, Lactobacillus ghanensis, Lactobacillus gigeriorum, Lactobacillus ginsenosidimutans, Lactobacillus gorillae, Lactobacillus graminis, Lactobacillus guizhouensis, Lactobacillushalophilus, Lactobacillus hammesii, Lactobacillus hamsteri, Lactobacillus harbinensis, Lactobacillus hayakitensis, Lactobacillus heilongjiangensis, Lactobacillus helsingborgensis, Lactobacillus helveticus, Lactobacillus herbarum, Lactobacillus heterohiochii, Lactobacillus hilgardii, Lactobacillus hokkaidonensis, Lactobacillus hominis, Lactobacillus homohiochii, Lactobacillus hordei, Lactobacillus iatae, Lactobacillus iners, Lactobacillus ingluviei, Lactobacillus insectis, Lactobacillus insicii, Lactobacillus intermedius, Lactobacillus intestinalis, Lactobacillus iwatensis, Lactobacillus ixorae, Lactobacillus japonicus, Lactobacillus jensenii, Lactobacillus johnsonii, Lactobacillus kalixensis, Lactobacillus kefiranofaciens, Lactobacillus kefiri, Lactobacillus kimbladii, Lactobacillus kimchicus, Lactobacillus kimchiensis, Lactobacillus kisonensis, Lactobacillus kitasatonis, Lactobacillus koreensis, Lactobacillus kullabergensis、Lactobacillus kunkeei、Lactobacillus larvae、Lactobacillus leichmannii、Lactobacillusletivazi, Lactobacillus lindneri, Lactobacillus malefermentans, Lactobacillus mali, Lactobacillus manihotivorans, Lactobacillus mellifer, Lactobacillus mellis, Lactobacillus melliventris, Lactobacillus micheneri, Lactobacillus mindensis, Lactobacillus mixtipabuli, Lactobacillus mobilis, Lactobacillus modestisalitolerans, Lactobacillus mucosae, Lactobacillus mudanjiangensis, Lactobacillus murinus, Lactobacillus nagelii, Lactobacillus namurensis, Lactobacillus nantensis, Lactobacillus nasuensis, Lactobacillus nenjiangensis, Lactobacillus nodensis, Lactobacillus odoratitofui, Lactobacillus oeni, Lactobacillus oligofermentans, Lactobacillus ori, Lactobacillus oryzae, Lactobacillus otakiensis, Lactobacillus ozensis, Lactobacillus panis, Lactobacillus pantheris, Lactobacillus parabrevis, Lactobacillus parabuchneri, Lactobacillus paracollinoides, Lactobacillus parafarraginis, Lactobacillus parakefiri, Lactobacillus paraalimentarius, Lactobacillus paraplantarum, Lactobacillus pasteurii, Lactobacillus paucivorans, Lactobacilluspentosus, Lactobacillus perolens, Lactobacillus plajomi, Lactobacillus plantarum, Lactobacillus pobuzihii, Lactobacillus pontis, Lactobacillus porcinae, Lactobacillus psittaci, Lactobacillus rapi, Lactobacillus rennanquilfy, Lactobacillus rennini, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus rodentium, Lactobacillus rogosae, Lactobacillus rossiae, Lactobacillus ruminis, Lactobacillus saerimneri, Lactobacillus sakei, Lactobacillus salivarius, Lactobacillus sanfranciscensis, Lactobacillus saniviri, Lactobacillus satsumensis, Lactobacillus secaliphilus, Lactobacillus selangorensis, Lactobacillus senioris, Lactobacillus senmaizukei, Lactobacillus sharpeae, Lactobacillus shenzhenensis, Lactobacillus sicerae, Lactobacillus silagei, Lactobacillus siliginis, Lactobacillus similis, Lactobacillus songhuajiangensis, Lactobacillus spicheri, Lactobacillus sucicola, Lactobacillus suebicus、Lactobacillus sunkii、Lactobacillus taiwanensis、Lactobacillus thailandensis、Lactobacillus tucceti、Lactobacillusultunensis, Lactobacillus uvarum, Lactobacillus vaccinostercus, Lactobacillus vaginalis, Lactobacillus vermiforme, Lactobacillus vespulae, Lactobacillus vini, Lactobacillus wasatchensis, Lactobacillus Examples of Lactobacillus bacteria include, but are not limited to, Lactobacillus xiangfangensis, Lactobacillus yonginensis, or Lactobacillus zymae. As used herein, it should be understood that "Lactobacillus bacteria" also refers to and encompasses Lactobacillus species that have recently been reclassified into alternative genera. These include Atopobium (e.g., A. deltae), Carnobacterium (e.g., C. alterfunditum), Weissella (e.g., W. beninensis), Oenococcus (e.g., O. oeni), Leuconostoc (e.g., L. carnosum), Pediococcus (e.g., P. acidilactici), Paralactobacillus (e.g., P. selangorensis), Holzapfelia (e.g., H. floricola), Amylolactobacillus (e.g., A. amylophilus), Bombilactobacillus (e.g., B. mellifer), Companilactobacillus (e.g., C. alimentarius), Lapidilactobacillus (e.g., L. concavus), Agrilact obacillus (e.g., A. composti), Schleiferilactobacillus (e.g., S. perolens), Loigolactobacillus (e.g., L. coryniformis), Lacticaseibacillus (e.g., L. casei), Latilactobacillus (e.g., L. sakei), Dellaglioa (e.g., D. algidus), Liquorilactobacillus (e.g., L. mail), Ligilactobacillus (e.g., L. salivarius), Lactiplantibacillus (e.g., L. plantarum), Furfurilactobacillus (e.g., F. rossiae), Paucilactobacillus (e.g., P. vaccinostercus), Limosilactobacillus (e.g., L.fermentum), Fructilactobacillus (e.g., F. fructivorans), Acetilactobacillus (e.g., A. jinshani), Apilactobacillus (e.g., A. kunkeei), Levilactobacillus (L. brevis), Secundilactobacillus (e.g., S. collinoides), and Lentilactobacillus (e.g., L. buchneri). It should be understood that the examples of recently reclassified Lactobacillus are provided for clarity only and are not intended to be limiting in any way. Thus, for example, Lactiplantibacillus is not limited to only L. plantarum, but rather includes any species of Lactiplantibacillus known or not yet known, or currently classified or not yet classified. Therefore, when referring to Lactiplantibacillus, the species include L. argentoratensis, L. daoliensis, L. daowaiensis, L. dongliensis, L. fabifermentans, L. garii, L. herba rum, L.modestisalitolerans, L.mudanjiangensis, L.paraplantarum, L.pentosus, L.pingfangensis, L.plajomi, L.plantarum, L.songbeiensis, and L.rum.xiangfangensis. Such understanding is applicable to all of the above genera. In some embodiments, the Lactobacillus bacterium is a wild-type natural isolate of Lactobacillus. In some embodiments, the Lactobacillus bacterium is a laboratory Lactobacillus strain that has undergone a breeding process of mutagenesis and selection. As used herein, "Lactobacillus bacterium" refers to any wild-type or laboratory strain of a known Lactobacillus bacterium. Furthermore, when referring to any specific Lactobacillus species, the recitation of the species also includes any wild-type or laboratory strain of a known Lactobacillus species.

[0048] "Encoding" refers to the inherent property of a particular sequence of nucleotides in a polynucleotide, such as a gene, cDNA, or mRNA, to serve as a template for the synthesis of other polymers and macromolecules in biological processes that have either a defined sequence of nucleotides (i.e., rRNA, tRNA, and mRNA) or a defined sequence of amino acids, and the biological properties resulting therefrom. Thus, a gene encodes a protein when the protein is produced in a cell or other biological system by transcription and translation of the mRNA corresponding to that gene. Both the coding strand, whose nucleotide sequence is identical to the mRNA sequence and is usually provided in a sequence listing, and the non-coding strand, which is used as a template for transcription of the gene or cDNA, can be said to encode the protein or other product of that gene or cDNA.

[0049] An "expression vector" refers to a vector containing a recombinant polynucleotide comprising an expression control sequence operably linked to a nucleotide sequence to be expressed. An expression vector contains sufficient cis-acting elements for expression; other elements for expression can be supplied by the host cell or in an in vitro expression system. Expression vectors include all expression vectors known in the art, such as cosmids, plasmids (e.g., naked or contained in liposomes), and viruses (e.g., Sendai virus, lentivirus, retrovirus, adenovirus, and adeno-associated virus) that incorporate the recombinant polynucleotide.

[0050] As used herein, "fitness" refers to the ability of bacteria to survive, grow, or otherwise be metabolically active in different environments. Fitness is generally measured by several methods, including measuring biofilm growth, survival in water, resistance to desiccation, and growth rate (e.g., doubling time). In the context of the present disclosure, fitness is generally measured via measuring growth rate. Furthermore, in the context of the present disclosure, the identity of a particular signal peptide, payload protein, or both, is a factor that can affect the fitness of a bacterial strain. If the use of a particular signal peptide or signal peptide and payload protein combination increases the doubling time of a bacterium, the bacterium is considered to be less fit. Similarly, if the use of a particular signal peptide or signal peptide and payload protein combination decreases the doubling time of a bacterium (i.e., the bacterium doubles at a faster rate), the bacterium is considered to be more fit. Changes in bacterial growth rate can be an indicator of how well the bacterium can survive in harsher environments, such as the acidic environment of the gut. If a particular signal peptide or signal peptide / payload protein combination increases the doubling time of bacteria, this may indicate that the use of that combination would be detrimental in a more severe growth environment. Conversely, if a particular signal peptide or signal peptide / payload protein combination decreases the doubling time of bacteria, this may indicate that the use of that combination has no effect on, or may even have a positive effect on, bacterial viability in a more severe growth environment.

[0051] As used herein, "fitness cost" refers to a change in bacterial growth rate as a result of the use of a particular signal peptide or signal peptide and payload protein combination. If the use of a particular signal peptide or signal peptide and payload protein combination increases the doubling time of the bacterium, that particular combination is considered to have a negative fitness cost. Similarly, if the use of a particular signal peptide or signal peptide and payload protein combination decreases the doubling time of the bacterium (i.e., the bacterium doubles at a faster rate), that particular combination is considered to have a positive fitness cost. For the purposes of this disclosure, an optimal signal peptide is one that strikes a balance between increased protein secretion and fitness cost.

[0052] As used herein, "genetic modification" or any grammatical variation thereof refers to the act of introducing a coding nucleic acid into a bacterial cell to promote expression of a recombinant protein within the cell. The nucleic acid can be DNA, mRNA, tRNA, or rRNA. The nucleic acid is composed of nucleotide monomers, and each triplet of monomers (codon) encodes either a triplet of RNA nucleotide monomers (if the nucleic acid is DNA) or an amino acid (if the nucleic acid is RNA). The DNA also contains one or more promoter regions that indicate where transcription of the DNA should begin. The mRNA also contains a ribosome binding site that indicates where translation of the mRNA should begin, as well as one or more stop codons that indicate where mRNA translation should end.

[0053] In any embodiment or aspect disclosed herein, the nucleic acid encoding the recombinant protein disclosed herein can be introduced into bacterial cells using any method known to those skilled in the art for such introduction. Such methods include transfection, transformation, transduction, infection (e.g., viral transduction), injection, microinjection, gene gun, nucleofection, nanoparticle irradiation, transformation, conjugation, applying nucleic acid in gel, oil, or cream, electroporation, using lipid-based transfection reagents, or any other suitable transfection method. Those skilled in the art can easily understand and adapt such methods using easily identifiable literature sources.

[0054] As used herein, the terms "transformation" and "transfection" are intended to refer to various art-recognized techniques for introducing exogenous nucleic acid into a host cell, including calcium phosphate or calcium chloride co-precipitation, DEAE-dextran-mediated transfection, lipofection (e.g., LIPOFECTIN® (Invitrogen Corp., San Diego, CA), LIPOFECTAMINE® (Invitrogen), FUGENE® (Roche Applied Science, Basel, Switzerland), JETPEI™ (Polyplus-transfection Inc., New York, NY), EFFECTENE® (Qiagen, Valencia, CA), DREAMFECT™ (OZ Biosciences, France), etc.), or electroporation (e.g., in vivo electroporation). Suitable methods for transforming or transfecting host cells are described in Sambrook, et al. (Molecular Cloning: A Laboratory Manual. 2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989), and other laboratory manuals.

[0055] Further methods and materials for non-viral delivery of nucleic acids into cells include particle bombardment, virosomes, liposomes, immunoliposomes, polycation or lipid-nucleic acid complexes, naked DNA, artificial virions, and drug-enhanced DNA uptake. Lipofection is described, for example, in U.S. Pat. Nos. 5,049,386, 4,946,787, and 4,897,355, and lipofection reagents are commercially available (e.g., TRANSFECTAM™ and LIPOFECTIN™). Cationic and neutral lipids suitable for efficient receptor-recognition lipofection of polynucleotides include those disclosed in WO91 / 17424 and WO91 / 16024.

[0056] The methods described herein include producing recombinant proteins in a microbial host. As used herein, recombinant refers to proteins or nucleic acids that may or may not naturally exist in the host, or that may or may not be produced by the host. For example, in some embodiments, the host may be a specific Lactobacillus species, and the recombinant protein expressed may also be naturally present in the specific Lactobacillus species. In another example, the host may be a specific Lactobacillus species, and the recombinant protein expressed may be one that does not naturally exist in the specific Lactobacillus species, such as a protein from a different Lactobacillus species or an entirely different organism.

[0057] As used herein, "payload protein" or "protein of interest" refers to a protein produced by a host and transported to the extracellular space via the secretory pathway, facilitated by the presence of a synthetic signal peptide. Upon secretion into the extracellular space, the payload protein may be fused to all, some, or none of the synthetic signal peptides. Optionally, the payload protein, still partially or completely attached to the synthetic signal peptide, may be further processed, e.g., to remove the remaining signal peptide. The payload protein may be any known or yet unknown protein, such as an enzyme, enzyme inhibitor, growth factor, hormone, antibody, antigen, vaccine, therapeutic agent, or any combination thereof. More specific examples are described herein below. The payload proteins secreted by the various genetically modified bacteria (interchangeably referred to as "modified bacteria") disclosed herein may be provided to a subject as a pharmaceutical composition. Additionally or alternatively, the modified bacteria themselves may be provided to a subject as a pharmaceutical composition.

[0058] In some embodiments, the payload protein or protein of interest comprises an antibody. "Antibody" is intended in a broad sense to include monoclonal antibodies, including murine, human, humanized, and chimeric monoclonal antibodies; antigen-binding fragments; multispecific antibodies, such as bispecific, trispecific, and tetraspecific antibodies; dimeric, tetrameric, or multimeric antibodies; single-chain antibodies; domain antibodies; and any other modified configuration of an immunoglobulin molecule that contains an antigen-binding site of the desired specificity. A "full-length antibody" consists of two heavy chains (HC) and two light chains (LC) interconnected by disulfide bonds, and multimers thereof (e.g., IgM). Each heavy chain consists of a heavy chain variable region (VH) and a heavy chain constant region (comprising domains CH1, hinge, CH2, and CH3). Each light chain consists of a light chain variable region (VL) and a light chain constant region (CL). The VH and VL regions can be further subdivided into regions of hypervariability called complementarity-specific regions (CDRs), interspersed with framework regions (FRs). Each VH and VL consists of three CDR and four FR segments, arranged from amino to carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. Depending on the amino acid sequence of the heavy-chain constant domain, immunoglobulins can be assigned to five major classes: IgA, IgD, IgE, IgG, and IgM. IgA and IgG are further classified as isotypes: IgA1, IgA2, IgG1, IgG2, IgG3, and IgG4. The "light chains" of antibodies of any vertebrate species can be assigned to one of two clearly distinct types, namely kappa (κ) and lambda (λ), based on the amino acid sequence of their constant domain.

[0059] In some embodiments, the payload protein or protein of interest comprises an antibody fragment. An "antibody fragment," "antigen-binding fragment," or "antigen-binding domain" refers to the portion of an immunoglobulin molecule that binds to an antigen. Antigen-binding fragments may be synthetic, enzymatically derived, or genetically engineered polypeptides, and include VH, VL, VH and VL, Fab, F(ab')2, Fd, and Fv fragments, domain antibodies (dAbs) consisting of one VH domain or one VL domain, shark variable IgNAR domains, camelized VH domains, amino acid residues mimicking the CDRs of an antibody, such as the FR3-CDR3-FR4 portion, HCDR1, HCDR2, and / or HCDR3, and LCDR1, LCDR2, and / or LCDR3. The VH and VL domains may be linked together via synthetic linkers to form various types of single chain antibody designs, where the VH / VL domains may pair intramolecularly, or, when the VH and VL domains are expressed by separate single chain antibody constructs, intermolecularly to form a monovalent antigen-binding site, such as a single chain Fv (scFv) or diabody. Other antibody fragments, or antigen-binding fragments, are known in the art, and any such fragments are applicable to the present disclosure.

[0060] The methods described herein involve producing recombinant proteins in bacterial hosts. As used herein, "recombinant protein" includes a payload protein and a synthetic signal peptide fused directly or indirectly thereto. As used herein, a signal peptide is any protein or peptide fused directly or indirectly to the N-terminus of a payload protein that promotes extracellular secretion of the payload protein after it is produced.

[0061] The methods disclosed herein utilize synthetic signal peptides to increase extracellular secretion of payload proteins by bacteria. As used herein, "synthetic signal peptide" refers to a recombinantly produced signal peptide whose sequence is generated as provided herein. A recombinantly produced signal peptide may be referred to as a "synthetic signal peptide" or simply a "signal peptide." In some embodiments, a signal peptide may comprise a synthetic pre-protein signal peptide. As emphasized above, the term synthetic in this context refers to a recombinantly produced pre-protein signal peptide (whose sequence is generated as provided herein). Hereinafter, a pre-signal peptide may be referred to as a "synthetic" pre-signal peptide or simply a pre-protein signal peptide.

[0062] A pre-protein signal peptide (synthetic or natural) comprises 10-50 amino acids that are added directly to the N-terminus of a payload protein or indirectly (e.g., using one or more spacers) to the N-terminus of a payload protein.

[0063] The synthetic pre-protein signal peptide can be attached to adjacent amino acids via attachment of the adjacent amino acids to the N-terminal amino acid, for example, by a peptide bond, a peptide spacer (e.g., LEISSTCDA represented by SEQ ID NO: 124), or a membrane-associated / lipophilic alpha-helical signal peptide (e.g., MISTIC represented by SEQ ID NO: 125).

[0064] The chemical makeup of peptides is described herein by a series of single-letter amino acid abbreviations, or "amino acid sequence(s)" or "sequence(s)," which are conventional and known to those of skill in the art. Although reference sequences are explicitly disclosed, in any aspect and embodiment, the reference sequence may be modified to include conservative amino acid substitutions, as well as variants and fragments, while maintaining the characteristics and functionality of the reference sequence.

[0065] As used herein, "substantially identical" or "substantially similar" means that a polypeptide or nucleic acid molecule exhibits at least 50% identity to a reference amino acid sequence (e.g., any one of the amino acid sequences described herein) or nucleic acid sequence (e.g., any one of the nucleic acid sequences described herein). Preferably, such sequences are at least 60%, more preferably 80% or 85%, and more preferably 90%, 95%, or even 99% identical at the amino acid level or nucleic acid level to the sequence used for comparison. These proteins may include homologs, orthologs, allelic variants, and functional variants. Typically, 50% or greater identity between two polypeptide sequences is considered to be an indication of functional equivalence. Identity between polypeptides is preferably determined by the Smith-Waterman homology search algorithm implemented in the MPSRCH program (Oxford Molecular) using an affine gap search with parameters of a gap opening penalty of -12 and a gap extension penalty of 1.

[0066] Sequence identity can be measured / determined using sequence analysis software (e.g., Sequence Analysis Software Package of the Genetics Computer Group, University of Wisconsin Biotechnology Center, 1710 University Avenue, Madison, Wis. 53705, BLAST, BESTFIT, GAP, or PILEUP / PRETTYBOX programs). Such software matches identical or similar sequences by assigning degrees of homology to various substitutions, deletions, and / or other modifications. Conservative substitutions typically include substitutions within the following groups: glycine, alanine, valine, isoleucine, leucine, aspartic acid, glutamic acid, asparagine, glutamine, serine, threonine, lysine, arginine, and phenylalanine, tyrosine. An exemplary method for measuring the degree of identity can use the BLAST program, where a probability score of e3 to e100 indicates closely related sequences. In some embodiments, sequence identity is determined using BLAST with default settings.

[0067] These proteins may contain one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) conservative amino acid substitutions, i.e., substitution of one amino acid for another amino acid with a related side chain, compared to the disclosed proteins. Genetically encoded amino acids are generally classified into four families: (1) acidic, i.e., aspartic acid, glutamic acid; (2) basic, i.e., lysine, arginine, histidine; (3) nonpolar, i.e., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan; and (4) uncharged polar, i.e., glycine, asparagine, glutamine, cysteine, serine, threonine, tyrosine. Phenylalanine, tryptophan, and tyrosine are sometimes collectively classified as aromatic amino acids. Typically, substitution of a single amino acid within these families does not significantly affect biological activity. The proteins can have one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) single amino acid deletions relative to the disclosed protein sequences. The proteins can also include one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) insertions (e.g., of 1, 2, 3, 4, or 5 amino acids each) relative to the disclosed protein sequences.

[0068] The compositions disclosed herein can be provided to a subject in various ways, such as by administering the composition to the subject.As used herein, administering or administering refers to providing a composition to a subject or providing a composition.Oral administration, as used herein, refers to the delivery of active agents via the mouth.Topical administration, as used herein, refers to the delivery of active agents to the surface of the body, such as the skin, mucous membranes (e.g., nasal membrane, vaginal membrane, buccal membrane, etc.).

[0069] The various compositions disclosed herein may be useful in the treatment of several diseases, such as cancer. As used herein, cancer refers to a pathological condition characterized by uncontrolled cell proliferation. Examples of cancer include, but are not limited to, squamous cell carcinoma, small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, gastrointestinal cancer, Hodgkin's lymphoma and non-Hodgkin's lymphoma, pancreatic cancer, glioblastoma, cervical cancer, colon cancer, colorectal cancer, endometrial cancer or uterine cancer, kidney cancer such as renal cell carcinoma and Wilms' tumor, basal cell carcinoma, melanoma, prostate cancer, and esophageal cancer.

[0070] The various compositions disclosed herein may also be useful for treating other indications. For example, the compositions disclosed herein may be useful for treating one or more of the following diseases or disorders: infection, functional bowel disorder, allergy, short bowel syndrome, chronic idiopathic constipation, COPD, inflammation, chronic inflammation, colorectal cancer, periodontitis, oral mucositis, infection, pathogenic E. coli, inflammatory bowel disease, ulcerative colitis, Crohn's disease, type 2 diabetes, obesity, MERS-CoV, SARS-CoV, bovine viral diarrhea, hypercholesterolemia, influenza, atopic dermatitis, or diseases or disorders that may result from any of the diseases or disorders described herein. Therefore, it should be understood that the list of diseases or disorders is not limited to the listed diseases or disorders, but also encompasses any disease or disorder derived therefrom. For example, inflammation and chronic inflammation are associated with many diseases or disorders. Therefore, the compositions provided herein that can be used to treat inflammation or chronic inflammation may also be useful for treating diseases or disorders derived therefrom.

[0071] The various compositions disclosed herein may contain one or more drugs, biologics, or active agents (which are used interchangeably herein and refer to chemical compounds or compounds that elicit a desired pharmacological or physiological effect), and may also contain therapeutically active, prophylactically active, or cosmetically active agents, i.e., payloads. "Drugs," "biologicals," and "active agents" include any pharmaceutically acceptable, pharmacologically active derivatives and analogs of the drugs, biologics, and active agents specifically mentioned herein, including, but not limited to, salts, esters, amides, prodrugs, active metabolites, inclusion complexes, analogs, and the like. Suitable drugs, biologics, and active agents include alcohol deterrents, amino acids, ammonia detoxifiers, anabolic agents, stimulants, analgesics, androgens, anesthetics, appetite reducing compounds, appetite suppressants, antagonists, antiallergics, antiamebics, antianemics, antianginals, antianxiety agents, antiarthritics, antiarteriosclerotics, antibacterials, anticancer agents including antineoplastic agents and anticancer boosters, anticholinergics, antigallstone agents, anticoagulants, antithrombotic agents ... Anti-insecticides, anticonvulsants, antidepressants, antidiabetics, antidiarrheals, antidiuretics, detoxifying agents, antidyskinesia agents, antiemetics, antiepileptics, antiestrogens, antifibrinolytics, antifungals, antiglaucoma agents, antihemophilia agents, antihemorrhagic agents, antihistamines, antihyperlipidemic agents, antihyperlipidemic agents, antihypertensive agents, anti-hypertensive agents, anti-infective agents such as antibiotics and antivirals, steroidal and non-steroidal anti-inflammatory agents, antikeratinizing agents, antimalarials antibacterial agents, antimigraine drugs, antimitotic agents, antifungal agents, antiemetics, antitumor agents, antineutropenic agents, antiobsessive-compulsive agents, antiparasitic agents, antiparkinsonian drugs, antipneumocystis pneumonia agents, antiproliferative agents, antiprostate hyperplasia agents, antiprotozoal agents, antipruritic agents, antipsoriatic agents, antipsychotic drugs, antipyretics, antispasmodics, antirheumatic drugs, antischistosomiasis drugs, antiseborrheic drugs, anticonvulsants, antithrombotic agents, antituberculous drugs, antitussives, antiulcer drugs, antiurinary tract stone agents, antiviral drugs , gastroesophageal reflux disease therapeutic agents, anti-anxiety agents, appetite suppressants, attention deficit disorder (ADD) and attention deficit hyperactivity disorder (ADHD) therapeutic agents, bacteriostatic and bactericidal agents, prostatic hyperplasia therapeutic agents, blood glucose regulators, bone resorption inhibitors, bronchodilators, carbonic anhydrase inhibitors, cardiovascular preparations such as antianginal agents, antiarrhythmic agents, beta-blockers, calcium channel blockers, cardiac depressants, cardiovascular therapeutic agents, cardioprotective agents, and cardiotonic agents, central nervous system (CNS) acting agents,Central nervous system stimulants, choleretics, cholinergics, acetylcholine receptor stimulants, cholinesterase inactivators, anticoccidials, cognitive aids and enhancers, cough and cold medicines (including decongestants), sedatives, diagnostic aids, diuretics, dopamine agonists, ectoparasiticides, emetics, enzymes that inhibit the formation of plaque, tartar, or dental caries, enzyme inhibitors, estrogens, fibrinolytics, fluoride anticaries / anticaries, free oxygen radical scavengers, gastrointestinal motility agents, genetic material, glucocorticoids, gonadotropins, hemostatic agents, herbal remedies, histamine H2 receptors receptor antagonists, hormones, hormone dissolving agents, hypnotics, cholesterol lowering agents, hypoglycemic agents, lipid lowering agents, blood pressure lowering agents, immunological agents, immunomodulators, immunoregulators, immunostimulants, immunosuppressants, impotence treatment aids, inhibitors, keratolytic agents, leukotriene inhibitors, liver damage treatment agents, metal chelating agents such as tetrasodium ethylenediaminetetraacetate, mitotic inhibitors, mood regulators, mucolytic agents, mucosal protective agents, muscle relaxants, mydriatics, narcotic antagonists, neuroleptics, neuromuscular blocking agents, neuroprotective agents, nicotine, NMDA antagonists, non-hormonal sterol derivatives, nutritional agents, e.g., bile acid, amines, essential amino acids and fatty acids, ophthalmic drugs such as those for treating glaucoma, uterotonics, analgesics, parasympatholytics, peptides, plasminogen activators, platelet activating factor antagonists, platelet aggregation inhibitors, post-stroke and post-head trauma treatments, enhancers, progestins, prostaglandins, prostate growth inhibitors, proteolytic enzymes as wound cleansers, prothyroid stimulating hormones, psychostimulants, psychotropic drugs, radioactive agents, regulating agents, relaxants, redistributing agents, scabicides, sclerosing agents, sedatives, hypnotics, selective adenosine A1 antagonists, serotonin antagonists, serotonin inhibitors anti-inflammatory agents, serotonin receptor antagonists, steroids such as progestogens, estrogens, corticosteroids, androgens, and anabolic agents, smoking cessation agents, stimulants, depressants, sympathomimetics, synergists, thyroid hormones, thyroid inhibitors, thyroid stimulants, tranquilizers, tooth desensitizers, tooth whitening agents, agents such as peroxides, metal chlorites, perborates, percarbonates, peracids, and combinations thereof, agents for treating unstable angina, uricosurics, vasoconstrictors, vasodilators including those for general coronary, peripheral, and cerebral blood vessels, wound treatment agents, wound healing agents,These may include, but are not limited to, xanthine oxidase inhibitors, etc.

[0072] Various compositions disclosed herein may contain a therapeutically effective amount of a drug, biological agent, or active agent. A therapeutically effective amount refers to a sufficient amount of a drug, biological agent, or active agent (alone or together with one or more other active agents) to induce a desired response, such as preventing, treating, alleviating, and / or ameliorating a condition. A therapeutically effective amount of an active agent, alone or together with one or more other active agents, can be determined in a number of different ways, such as by assaying for the reduction of one or more signs or symptoms associated with the condition of interest or by measuring the levels of one or more molecules associated with the condition being treated.

[0073] The term "subject" includes living organisms (e.g., mammals), including those in which an immune response can be elicited. As used herein, the terms "subject" or "individual" can be used interchangeably. As used herein, "subject" refers to a human or non-human mammal. Non-human mammals include, for example, mammalian livestock and pets, such as sheep, cows, pigs, dogs, non-human primates, cats, and mice. In some embodiments, the subject is a human.

[0074] As used herein, the phrase "pharmaceutically acceptable" refers to compounds, materials, compositions and / or dosage forms that are suitable for use in contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication, within the scope of sound medical judgment and commensurate with a reasonable benefit / risk ratio.

[0075] As used herein, the phrase "in need thereof" means that a subject (animal or mammal) has been identified as having a need for a particular method or treatment. In some embodiments, identification can be by any diagnostic means. The animal or mammal may be in need of any of the methods and treatments described herein. In some embodiments, the animal or mammal is in or will be in an environment where a particular disease, disorder, or condition is prevalent.

[0076] As used herein, the term "therapeutic" refers to treatment and / or prophylaxis. A therapeutic effect is achieved by suppressing, ameliorating, or eradicating a disease condition.

[0077] "Treating" a disease, as that term is used herein, means reducing the frequency or severity of at least one sign or symptom of the disease or disorder experienced by a subject.

[0078] Ranges: Throughout this disclosure, various aspects of the embodiments may be expressed in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation. Accordingly, the description of a range should be considered to have specifically disclosed all possible subranges and individual numerical values ​​within that range. For example, the description of a range such as 1 to 6 should be considered to have specifically disclosed subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., and individual numerical values ​​within that range, e.g., 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range. Unless expressly stated to the contrary, a disclosed range includes the endpoints of the range.

[0079] Synthetic Preprotein Signal Peptides Provided herein are synthetic preprotein signal peptides that increase secretion of payload proteins from bacteria. Tables 1 and 2 below list the various amino acid sequences and polynucleotide sequences, respectively, referred to herein. [Table 1-1] [Table 1-2] [Table 1-3] [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5]

[0080] The synthetic pre-protein signal peptides disclosed herein have been optimized for use in Lactobacillus bacteria, and any bacterium within the genus Lactobacillus can be used to express the payload proteins provided herein. Specific examples of suitable bacterial species are provided herein below to illustrate the specific synthetic signal peptides developed.

[0081] In some embodiments, a pre-protein signal peptide is provided. In some embodiments, the pre-protein signal peptide comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence shown in Table 1. In some embodiments, the pre-protein signal peptide comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence shown in Table 1. In some embodiments, the pre-protein signal peptide comprises an amino acid sequence having at least 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence shown in Table 1. In some embodiments, the pre-protein signal peptide comprises an amino acid sequence having at least 95% identity to an amino acid sequence shown in Table 1. In some embodiments, the pre-protein signal peptide comprises an amino acid sequence having at least 98% identity to an amino acid sequence shown in Table 1. In some embodiments, the pre-protein signal peptide comprises an amino acid sequence shown in Table 1.

[0082] In some embodiments, the pre-protein signal peptide comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:148, or SEQ ID NO:180. In some embodiments, the pre-protein signal peptide comprises an amino acid sequence having at least 98% identity to an amino acid sequence selected from SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:148, or SEQ ID NO:180. In some embodiments, the preprotein signal peptide comprises an amino acid sequence selected from SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:148, or SEQ ID NO:180.

[0083] In some embodiments, the pre-protein signal peptide is SEQ ID NO:144, SEQ ID NO:145, SEQ ID NO:146, SEQ ID NO:147, SEQ ID NO:148, SEQ ID NO:149, SEQ ID NO:150, SEQ ID NO:151, SEQ ID NO:152, SEQ ID NO:153, SEQ ID NO:154, SEQ ID NO:155, SEQ ID NO:156, SEQ ID NO:157, SEQ ID NO:158, SEQ ID NO:159, SEQ ID NO:160, SEQ ID NO:161, SEQ ID NO:162, SEQ ID NO:163, SEQ ID NO:164, SEQ ID NO:165, SEQ ID NO:166, SEQ ID NO:167, SEQ ID NO:168, SEQ ID NO:169, SEQ ID NO:170, SEQ ID NO:171, SEQ ID NO:172, SEQ ID NO:173, SEQ ID NO:174, SEQ ID NO:175, SEQ ID NO:176, SEQ ID NO:177, SEQ ID NO:178, SEQ ID NO:179, SEQ ID NO:180, SEQ ID NO:181, SEQ ID NO:182, SEQ ID NO:183, SEQ ID NO:184, SEQ ID NO:185, SEQ ID NO:186, SEQ ID NO:187, SEQ ID NO:188, SEQ ID NO:189, SEQ ID NO:190, SEQ ID NO:191, SEQ ID NO:192, SEQ ID NO:193, SEQ ID NO:194, SEQ ID NO:195, SEQ ID NO:196, SEQ ID NO:197, SEQ ID NO:198, SEQ ID NO: 2, comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:183, SEQ ID NO:184, SEQ ID NO:185, SEQ ID NO:186, SEQ ID NO:187, SEQ ID NO:188, SEQ ID NO:189, SEQ ID NO:190, SEQ ID NO:191, SEQ ID NO:192, SEQ ID NO:193, SEQ ID NO:194, SEQ ID NO:195, SEQ ID NO:196, SEQ ID NO:197, SEQ ID NO:198, SEQ ID NO:199, SEQ ID NO:200, SEQ ID NO:201, SEQ ID NO:202, SEQ ID NO:203, SEQ ID NO:204, SEQ ID NO:205, SEQ ID NO:206, SEQ ID NO:207, SEQ ID NO:208, SEQ ID NO:209, SEQ ID NO:210, SEQ ID NO:211, or SEQ ID NO:212.In some embodiments, the pre-protein signal peptide is SEQ ID NO:144, SEQ ID NO:145, SEQ ID NO:146, SEQ ID NO:147, SEQ ID NO:148, SEQ ID NO:149, SEQ ID NO:150, SEQ ID NO:151, SEQ ID NO:152, SEQ ID NO:153, SEQ ID NO:154, SEQ ID NO:155, SEQ ID NO:156, SEQ ID NO:157, SEQ ID NO:158, SEQ ID NO:159, SEQ ID NO:160, SEQ ID NO:161, SEQ ID NO:162, SEQ ID NO:163, SEQ ID NO:164, SEQ ID NO:165, SEQ ID NO:166, SEQ ID NO:167, SEQ ID NO:168, SEQ ID NO:169, SEQ ID NO:170, SEQ ID NO:171, SEQ ID NO:172, SEQ ID NO:173, SEQ ID NO:174, SEQ ID NO:175, SEQ ID NO:176, SEQ ID NO:177, SEQ ID NO:178, 179, SEQ ID NO:180, SEQ ID NO:181, SEQ ID NO:182, SEQ ID NO:183, SEQ ID NO:184, SEQ ID NO:185, SEQ ID NO:186, SEQ ID NO:187, SEQ ID NO:188, SEQ ID NO:189, SEQ ID NO:190, SEQ ID NO:191, SEQ ID NO:192, SEQ ID NO:193, SEQ ID NO:194, SEQ ID NO:195, SEQ ID NO:196, SEQ ID NO:197, SEQ ID NO:198, SEQ ID NO:199, SEQ ID NO:200, SEQ ID NO:201, SEQ ID NO:202, SEQ ID NO:203, SEQ ID NO:204, SEQ ID NO:205, SEQ ID NO:206, SEQ ID NO:207, SEQ ID NO:208, SEQ ID NO:209, SEQ ID NO:210, SEQ ID NO:211, or SEQ ID NO:212.In some embodiments, the pre-protein signal peptide is SEQ ID NO:144, SEQ ID NO:145, SEQ ID NO:146, SEQ ID NO:147, SEQ ID NO:148, SEQ ID NO:149, SEQ ID NO:150, SEQ ID NO:151, SEQ ID NO:152, SEQ ID NO:153, SEQ ID NO:154, SEQ ID NO:155, SEQ ID NO:156, SEQ ID NO:157, SEQ ID NO:158, SEQ ID NO:159, SEQ ID NO:160, SEQ ID NO:161, SEQ ID NO:162, SEQ ID NO:163, SEQ ID NO:164, SEQ ID NO:165, SEQ ID NO:166, SEQ ID NO:167, SEQ ID NO:168, SEQ ID NO:169, SEQ ID NO:170, SEQ ID NO:171, SEQ ID NO:172, SEQ ID NO:173, SEQ ID NO:174, SEQ ID NO:175, SEQ ID NO:176, SEQ ID NO:177 , SEQ ID NO:178, SEQ ID NO:179, SEQ ID NO:180, SEQ ID NO:181, SEQ ID NO:182, SEQ ID NO:183, SEQ ID NO:184, SEQ ID NO:185, SEQ ID NO:186, SEQ ID NO:187, SEQ ID NO:188, SEQ ID NO:189, SEQ ID NO:190, SEQ ID NO:191, SEQ ID NO:192, SEQ ID NO:193, SEQ ID NO:194, SEQ ID NO:195, SEQ ID NO:196, SEQ ID NO:197, SEQ ID NO:198, SEQ ID NO:199, SEQ ID NO:200, SEQ ID NO:201, SEQ ID NO:202, SEQ ID NO:203, SEQ ID NO:204, SEQ ID NO:205, SEQ ID NO:206, SEQ ID NO:207, SEQ ID NO:208, SEQ ID NO:209, SEQ ID NO:210, SEQ ID NO:211, or SEQ ID NO:212.

[0084] In some embodiments, the pre-protein signal peptide comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 6. In some embodiments, the pre-protein signal peptide comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 6. In some embodiments, the pre-protein signal peptide comprises the amino acid sequence of SEQ ID NO: 6.

[0085] In some embodiments, the pre-protein signal peptide comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 7. In some embodiments, the pre-protein signal peptide comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 7. In some embodiments, the pre-protein signal peptide comprises the amino acid sequence of SEQ ID NO: 7.

[0086] In some embodiments, the preprotein signal peptide comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 14. In some embodiments, the preprotein signal peptide comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 14. In some embodiments, the preprotein signal peptide comprises the amino acid sequence of SEQ ID NO: 14.

[0087] In some embodiments, the preprotein signal peptide comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 17. In some embodiments, the preprotein signal peptide comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 17. In some embodiments, the preprotein signal peptide comprises the amino acid sequence of SEQ ID NO: 17.

[0088] In some embodiments, the preprotein signal peptide comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 18. In some embodiments, the preprotein signal peptide comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 18. In some embodiments, the preprotein signal peptide comprises the amino acid sequence of SEQ ID NO: 18.

[0089] In some embodiments, the pre-protein signal peptide comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 19. In some embodiments, the pre-protein signal peptide comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 19. In some embodiments, the pre-protein signal peptide comprises the amino acid sequence of SEQ ID NO: 19.

[0090] In some embodiments, the preprotein signal peptide comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 20. In some embodiments, the preprotein signal peptide comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 20. In some embodiments, the preprotein signal peptide comprises the amino acid sequence of SEQ ID NO: 20.

[0091] In some embodiments, the pre-protein signal peptide comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 47. In some embodiments, the pre-protein signal peptide comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 47. In some embodiments, the pre-protein signal peptide comprises the amino acid sequence of SEQ ID NO: 47.

[0092] In some embodiments, the preprotein signal peptide comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 48. In some embodiments, the preprotein signal peptide comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 48. In some embodiments, the preprotein signal peptide comprises the amino acid sequence of SEQ ID NO: 48.

[0093] In some embodiments, the preprotein signal peptide comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 148. In some embodiments, the preprotein signal peptide comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 148. In some embodiments, the preprotein signal peptide comprises the amino acid sequence of SEQ ID NO: 148.

[0094] In some embodiments, the preprotein signal peptide comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 180. In some embodiments, the preprotein signal peptide comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 180. In some embodiments, the preprotein signal peptide comprises the amino acid sequence of SEQ ID NO: 180. Polypeptides

[0095] In some embodiments, a polypeptide is provided, in some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide and Z1 is a payload protein.

[0096] In some embodiments, X1 comprises a preprotein signal peptide provided herein. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in Table 1. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to an amino acid sequence set forth in Table 1. In some embodiments, X1 comprises an amino acid sequence set forth in Table 1. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:148, or SEQ ID NO:180. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to an amino acid sequence selected from SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:148, or SEQ ID NO:180. In some embodiments, X1 comprises an amino acid sequence selected from SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:148, or SEQ ID NO:180.

[0097] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 6. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 6. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 6.

[0098] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 7. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 7. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 7.

[0099] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 14. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 14. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14.

[0100] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 17. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 17. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 17.

[0101] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 18. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 18. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 18.

[0102] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 19. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 19. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 19.

[0103] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 20. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 20. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 20.

[0104] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 47. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 47. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 47.

[0105] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 48. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 48. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 48.

[0106] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 148. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 148. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 148.

[0107] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 180. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 180. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 180.

[0108] In some embodiments, Z1 is a protein. In some embodiments, Z1 is a therapeutic protein. In some embodiments, Z1 is a payload protein selected from an antimicrobial peptide, a cytokine, an enzyme, a food protein, a growth factor, a hormone, an immunoreceptor, a mucolytic agent, an antibody, a nanobody, or a peptide. In some embodiments, Z1 is a payload protein selected from the group consisting of an antimicrobial peptide, a catestatin, a CRAMP, hBD1, hBD3, β-catethritin, dermaseptin, IL-10, IL-22, ClbS, TIGIT, TFF1, TFF2, TFF3, an anti-CCL2 antibody, an anti-CCL2 antibody fragment, an anti-IL23 antibody, an anti-IL23 antibody fragment, an anti-TNFα antibody, an anti-TNFα antibody fragment, GLP2, IGF1, leptin, and casein. In some embodiments, Z1 is a payload protein selected from the group consisting of an anti-TNFα antibody, an anti-TNFα antibody fragment, IL-10, IL-22, hBD3, leptin, and β-casein. In some embodiments, Z1 is an anti-TNFα antibody. In some embodiments, Z1 is an anti-TNFα antibody fragment. In some embodiments, Z1 is IL-10. In some embodiments, Z1 is IL-22. In some embodiments, Z1 is hBD3. In some embodiments, Z1 is leptin. In some embodiments, Z1 is β-casein. In some embodiments, Z1 is a protein comprising an amino acid sequence listed in Table 3 below. [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4] [Table 3-5] [Table 3-6] [Table 3-7] [Table 3-8] [Table 3-9] [Table 3-10] [Table 3-11]

[0109] It should be understood that the sequences shown in Table 3 are exemplary and are not intended to be limiting in any way. For example, SEQ ID NO:59 and SEQ ID NO:92 each contain the following subsequence: TPQNITDLCAEYHNTQIYTLNDKIFSYTESLAGKREMAIITFKNGAIFQVEVPGSQHIDSQKKAIERMKDTLRIAYLTEAKVEKLCVWNNKTPHAIAAISMANGPGPKKRKSV (SEQ ID NO: 143) This sequence promotes epithelial cell uptake. In some embodiments, the sequences shown in Table 2 may further comprise the sequence of SEQ ID NO: 143 at the N-terminus of the payload protein. In some embodiments, the amino acid sequence of Z1 is substantially similar to an amino acid sequence set forth in Table 3. In some embodiments, the amino acid sequence of Z1 is a fragment of an amino acid sequence set forth in Table 3. In some embodiments, the amino acid sequence of Z1 is substantially similar to or a fragment of an amino acid sequence set forth in Table 3, and further comprises the sequence of SEQ ID NO: 143 at the N-terminus of the payload protein. In some embodiments, the amino acid sequence of Z1 has at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in Table 3. Furthermore, it is understood that identity of Z1 can include, for example, amino acid sequences that are homologs of sequences set forth in Table 3. For example, Table 3 can provide amino acid sequences that correspond to human amino acid sequences. It is understood that the present embodiments are not limited to the human amino acid sequences provided, but may also include amino acid sequences of homologous proteins from, for example, non-human primates, mice, rats, dogs, horses, sheep, goats, etc.

[0110] In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in Table 1, and Z1 is a payload protein having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in Table 3. In some embodiments, X1 is as provided herein and Z1 is as provided herein.

[0111] In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a vaccine antigen. In some embodiments, the vaccine antigen is a bacterial antigen or a viral antigen. In some embodiments, the vaccine antigen is a bacterial antigen. In some embodiments, the bacterial antigen is a protein derived from a pathogenic bacterium. In some embodiments, the bacterial antigen is derived from Streptococcus agalactiae GBS, Borrelia burgdorferi, Colstridium tetani, Erysipelothrix rhusiopathiae, Helicobacter pylori, Salmonella enterica, Streptococcus mutans, Streptococcus pyogens, Streptococcus pneumonia, or Listeria monocytogenes. In some embodiments, the bacterial antigen comprises a pilus protein, Borellia burgdorferi outer surface protein A (OspA), tetanus toxin (TTFC), surface protective antigen A (SpaA), urease B (UreB), Cag12, flagellin (FliC), S. mutans serotype c (PAc), M6, pneumococcal surface attachment A (PsaA), pneumococcal protective protein A (PppA), listeriolysin O (LLO), or a fragment thereof. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a bacterial vaccine antigen comprising a pilus, OspA, TTFC, SpaA, UreB, Cag12, FliC, PAc, M6, PsaA, PppA, LLO, or a fragment thereof.

[0112] In some embodiments, the vaccine antigen is a viral antigen. In some embodiments, the viral antigen is derived from bovine coronavirus, coronavirus, dengue virus serotype 2, HIV-1, HPV-16, avian influenza, infectious bursal disease virus (IBDV), chicken anemia virus (CAV), porcine parvovirus, rotavirus, or hepatitis B virus. In some embodiments, the viral antigen comprises NSP4, SARS glycoprotein, SARS spike glycoprotein S, dengue virus envelope glycoprotein domain III (EDIII), V3 of HIV-1 gp120 glycoprotein, HPV-16 E7 protein, HPV-16 E6 protein, HPV-16 L1 protein, H5N1 avian influenza HA envelope protein, IBDV VP2, IBDV VP3, CAV VP1, porcine parvovirus VP2, rotavirus VP7, hepatitis B PreS domain, or hepatitis B PreSa domain. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a viral antigen comprising NSP4, a SARS glycoprotein, a SARS spike glycoprotein S, a dengue virus envelope glycoprotein domain III (EDIII), V3 of the HIV-1 gp120 glycoprotein, an HPV-16 E7 protein, an HPV-16 E6 protein, an HPV-16 L1 protein, an H5N1 avian influenza HA envelope protein, an IBDV VP2, an IBDV VP3, a CAV VP1, a porcine parvovirus VP2, a rotavirus VP7, a hepatitis B PreS domain, or a hepatitis B PreSa domain.

[0113] In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is an antimicrobial peptide. In some embodiments, the antimicrobial peptide is any antimicrobial peptide. In some embodiments, the antimicrobial peptide is hBD1, hBD2, hBD3, b-catethritin, dermaseptin, HD5, Reg3a, or C5a. In some embodiments, the antimicrobial peptide comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:23, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, or SEQ ID NO:55. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is an antimicrobial peptide comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:23, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, or SEQ ID NO:55.

[0114] In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is an enzyme. In some embodiments, the enzyme is any enzyme. In some embodiments, the enzyme is Clbs, shP28GST, Clbs (Gilliamella apicola), Clbs (Snodgrassella alvi), phytase, SUC2, SUIS, AnPep, β-galactosidase (Lactobacillus), LACPL exo-α-1,4-glucosidase, β-galactosidase (yeast), ACE2, xylose isomerase (RainFlu), xylose isomerase (Lactobacillus), MaIL, apyrase, or DL-endopeptidase. In some embodiments, the enzyme comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:74, SEQ ID NO:75, SEQ ID NO:76, SEQ ID NO:77, or SEQ ID NO:78. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is an enzyme comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:74, SEQ ID NO:75, SEQ ID NO:76, SEQ ID NO:77, or SEQ ID NO:78.

[0115] In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a food protein. In some embodiments, the food protein is any food protein. In some embodiments, the food protein is β-casein, α-casein, an α-casein-derived peptide, Ara h1, or Ara h2. In some embodiments, the food protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:89, or SEQ ID NO:90. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide described herein and Z1 is a food protein comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:89, or SEQ ID NO:90.

[0116] In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a growth factor. In some embodiments, the growth factor is any growth factor. In some embodiments, the growth factor is selected from IGF1, CTB-IGF1, or elafin. In some embodiments, the growth factor comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:91, SEQ ID NO:92, or SEQ ID NO:93. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a growth factor comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:91, SEQ ID NO:92, or SEQ ID NO:93.

[0117] In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is an immunoreceptor. In some embodiments, the immunoreceptor is any immunoreceptor. In some embodiments, the immunoreceptor is TIGIT. In some embodiments, the immunoreceptor comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 104. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is an immunoreceptor comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 104.

[0118] In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a mucolytic agent. In some embodiments, the mucolytic agent is any mucolytic agent. In some embodiments, the mucolytic agent is DNAse 1. In some embodiments, the mucolytic agent comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 105. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a mucolytic agent comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 105.

[0119] In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a peptide. In some embodiments, the peptide is any peptide. In some embodiments, the peptide is selected from adrenomedullin or urmin. In some embodiments, the peptide comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 121 or SEQ ID NO: 122. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a peptide comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:121 or SEQ ID NO:122.

[0120] In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a filament-binding protein. In some embodiments, the filament-binding protein is any filament-binding protein. In some embodiments, the filament-binding protein is filaggrin. In some embodiments, the filament-binding protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 123. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a filament-associated protein comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 123.

[0121] In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:20, and Z1 comprises a lectin. In some embodiments, the lectin is any lectin. In some embodiments, the lectin is TFF1, TFF2, or TFF3. In some embodiments, the cytokine comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:56, SEQ ID NO:57, or SEQ ID NO:58. In some embodiments, the polypeptide comprises a formula of X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 comprises a lectin comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:56, SEQ ID NO:57, or SEQ ID NO:58.

[0122] In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 comprises an amino acid having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:14, and Z1 comprises a nanobody. In some embodiments, the nanobody can be any nanobody. In some embodiments, the nanobody binds to a target protein, in which case the target protein can be any protein. In some embodiments, the nanobody binds to a target protein, in which case the target protein is TNFα, CCL2, IL23, EGFR-III, GLP2, VEGFB, ILR6, MERS, SARS, BVD, norovirus, RSV, TcdA, TcdB, PCSK9, Pseudomonas toxin, or IgE. In some embodiments, the Nanobody comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% to an amino acid sequence selected from SEQ ID NO:21, SEQ ID NO:106, SEQ ID NO:107, SEQ ID NO:108, SEQ ID NO:109, SEQ ID NO:110, SEQ ID NO:111, SEQ ID NO:112, SEQ ID NO:113, SEQ ID NO:114, SEQ ID NO:115, SEQ ID NO:116, SEQ ID NO:117, SEQ ID NO:118, SEQ ID NO:119, SEQ ID NO:120, SEQ ID NO:126, SEQ ID NO:127, SEQ ID NO:128, SEQ ID NO:129, SEQ ID NO:130, or SEQ ID NO:131.In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:14, and Z1 is selected from SEQ ID NO:21, SEQ ID NO:106, SEQ ID NO:107, SEQ ID NO:108, SEQ ID NO:109, SEQ ID NO:110, SEQ ID NO:111, SEQ ID NO:112. or an amino acid sequence that has at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% to an amino acid sequence selected from SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130, or SEQ ID NO: 131.

[0123] In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 comprises an amino acid having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:148, or SEQ ID NO:180, and Z1 comprises a hormone. In some embodiments, the hormone is any hormone. In some embodiments, the hormone is GLP1, exenatide-4, catestatin, leptin, nesfatin, linaclotide, plecanatide, uroguanylin, PYY, guanylin, or LCRF. In some embodiments, the hormone comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:25, SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:97, SEQ ID NO:98, SEQ ID NO:98, SEQ ID NO:99, SEQ ID NO:100, SEQ ID NO:101, SEQ ID NO:102, or SEQ ID NO:103. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises amino acids having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:148, or SEQ ID NO:180, and Z1 comprises a hormone comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:25, SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:97, SEQ ID NO:98, SEQ ID NO:99, SEQ ID NO:100, SEQ ID NO:101, SEQ ID NO:102, or SEQ ID NO:103.

[0124] In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:20, SEQ ID NO:148, or SEQ ID NO:180, and Z1 comprises a cytokine. In some embodiments, the cytokine is any cytokine. In some embodiments, the cytokine is IL-10, IL-22, or CTB-IL-22. In some embodiments, the cytokine comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:59, SEQ ID NO:282, SEQ ID NO:283, SEQ ID NO:284, or SEQ ID NO:285. In some embodiments, the polypeptide comprises a cytokine comprising the formula of X1-Z1, where X1 comprises amino acids having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:20, SEQ ID NO:148, or SEQ ID NO:180, and Z1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:59, SEQ ID NO:282, SEQ ID NO:283, SEQ ID NO:284, or SEQ ID NO:285.

[0125] In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 6, and Z1 comprises an anti-TNF-α antibody or an anti-TNF-α antibody fragment. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 6, and Z1 comprises an anti-TNF-α antibody or an anti-TNF-α antibody fragment. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 6, and Z1 comprises an anti-TNF-α antibody or an anti-TNF-α antibody fragment. In some embodiments, Z1 comprises an anti-TNF-α antibody. In some embodiments, Z1 comprises an anti-TNFα antibody fragment. In some embodiments, the anti-TNFα antibody is as provided herein. In some embodiments, the anti-TNFα antibody fragment is an anti-TNFα antibody fragment provided herein.

[0126] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:6, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:6, and Z1 comprises the amino acid of SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO:6, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO:6, and Z1 comprises amino acids of SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:6, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:6, and Z1 comprises amino acids of SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof.In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:6 and Z1 comprises amino acids of SEQ ID NO:21, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:6 and Z1 comprises amino acids of SEQ ID NO:118, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:6 and Z1 comprises amino acids of SEQ ID NO:119, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:6 and Z1 comprises amino acids of SEQ ID NO:120, or a fragment thereof.

[0127] In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:7, and Z1 comprises an anti-TNF-α antibody or an anti-TNF-α antibody fragment. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO:7, and Z1 comprises an anti-TNF-α antibody or an anti-TNF-α antibody fragment. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 comprises an amino acid sequence having the amino acid sequence of SEQ ID NO:7, and Z1 comprises an anti-TNF-α antibody or an anti-TNF-α antibody fragment. In some embodiments, Z1 comprises an anti-TNF-α antibody. In some embodiments, Z1 comprises an anti-TNFα antibody fragment. In some embodiments, the anti-TNFα antibody is as provided herein. In some embodiments, the anti-TNFα antibody fragment is an anti-TNFα antibody fragment provided herein.

[0128] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:7, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:7, and Z1 comprises the amino acid of SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO:7, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO:7, and Z1 comprises amino acids of SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7, and Z1 comprises amino acids of SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof.In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7 and Z1 comprises amino acids of SEQ ID NO:21, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7 and Z1 comprises amino acids of SEQ ID NO:118, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7 and Z1 comprises amino acids of SEQ ID NO:119, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7 and Z1 comprises amino acids of SEQ ID NO:120, or a fragment thereof.

[0129] In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 14, and Z1 comprises an anti-TNF-α antibody or an anti-TNF-α antibody fragment. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 14, and Z1 comprises an anti-TNF-α antibody or an anti-TNF-α antibody fragment. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 14, and Z1 comprises an anti-TNF-α antibody or an anti-TNF-α antibody fragment. In some embodiments, Z1 comprises an anti-TNF-α antibody. In some embodiments, Z1 comprises an anti-TNFα antibody fragment. In some embodiments, the anti-TNFα antibody is as provided herein. In some embodiments, the anti-TNFα antibody fragment is an anti-TNFα antibody fragment provided herein.

[0130] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 14, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 21, SEQ ID NO: 118, SEQ ID NO: 119, or SEQ ID NO: 120, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 14, and Z1 comprises the amino acid sequence of SEQ ID NO: 21, SEQ ID NO: 118, SEQ ID NO: 119, or SEQ ID NO: 120, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 14, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 21, SEQ ID NO: 118, SEQ ID NO: 119, or SEQ ID NO: 120, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 14, and Z1 comprises the amino acid sequence of SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14, and Z1 comprises the amino acid sequence of SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof.In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14 and Z1 comprises amino acids of SEQ ID NO: 21, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14 and Z1 comprises amino acids of SEQ ID NO: 118, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14 and Z1 comprises amino acids of SEQ ID NO: 119, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14 and Z1 comprises amino acids of SEQ ID NO: 120, or a fragment thereof.

[0131] In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 17, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 17, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 17, and Z1 comprises leptin, or a fragment thereof. In some embodiments, Z1 comprises leptin. In some embodiments, Z1 comprises a fragment of leptin.

[0132] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 17, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 17, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 17, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 17, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 17 and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 17 and Z1 comprises the amino acids of SEQ ID NO: 25, or a fragment thereof.

[0133] In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 18, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 18, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 18, and Z1 comprises leptin, or a fragment thereof. In some embodiments, Z1 comprises leptin. In some embodiments, Z1 comprises a fragment of leptin.

[0134] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 18, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 18, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 18, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 18, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 18 and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 18 and Z1 comprises the amino acids of SEQ ID NO: 25, or a fragment thereof.

[0135] In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 19, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 19, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 19, and Z1 comprises leptin, or a fragment thereof. In some embodiments, Z1 comprises leptin. In some embodiments, Z1 comprises a fragment of leptin.

[0136] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 19, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 19, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 19, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 19, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 19 and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 19 and Z1 comprises the amino acids of SEQ ID NO: 25, or a fragment thereof.

[0137] In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 47, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 47, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 47, and Z1 comprises leptin, or a fragment thereof. In some embodiments, Z1 comprises leptin. In some embodiments, Z1 comprises a fragment of leptin.

[0138] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 47, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 47, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 47, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 47, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:47 and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:47 and Z1 comprises the amino acids of SEQ ID NO:25, or a fragment thereof.

[0139] In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 48, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 48, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 48, and Z1 comprises leptin, or a fragment thereof. In some embodiments, Z1 comprises leptin. In some embodiments, Z1 comprises a fragment of leptin.

[0140] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 48, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 48, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 48, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 48, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:48 and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:48 and Z1 comprises the amino acids of SEQ ID NO:25, or a fragment thereof.

[0141] In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises leptin, or a fragment thereof. In some embodiments, Z1 comprises leptin. In some embodiments, Z1 comprises a fragment of leptin.

[0142] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 148 and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 148 and Z1 comprises the amino acids of SEQ ID NO: 25, or a fragment thereof.

[0143] In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 180, and Z1 comprises leptin, or a fragment thereof. In some embodiments, Z1 comprises leptin. In some embodiments, Z1 comprises a fragment of leptin.

[0144] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 180 and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 180 and Z1 comprises the amino acids of SEQ ID NO: 25, or a fragment thereof.

[0145] In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 20, and Z1 comprises IL-10 or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 20, and Z1 comprises IL-10 or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 20, and Z1 comprises IL-10 or a fragment thereof. In some embodiments, Z1 comprises IL-10. In some embodiments, Z1 comprises a fragment of IL-10.

[0146] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:20, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:24, SEQ ID NO:283, SEQ ID NO:284, or SEQ ID NO:285, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:20, and Z1 comprises the amino acid sequence of SEQ ID NO:24, SEQ ID NO:283, SEQ ID NO:284, or SEQ ID NO:285, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO:20, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO:24, SEQ ID NO:283, SEQ ID NO:284, or SEQ ID NO:285, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence at least 98% identical to the amino acid sequence of SEQ ID NO:20, and Z1 comprises the amino acid sequence of SEQ ID NO:24, SEQ ID NO:283, SEQ ID NO:284, or SEQ ID NO:285, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:20, and Z1 comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:24, SEQ ID NO:283, SEQ ID NO:284, or SEQ ID NO:285, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:20, and Z1 comprises the amino acid sequence of SEQ ID NO:24, SEQ ID NO:283, SEQ ID NO:284, or SEQ ID NO:285, or a fragment thereof.In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:20 and Z1 comprises the amino acid sequence of SEQ ID NO:24. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:20 and Z1 comprises the amino acid sequence of SEQ ID NO:283. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:20 and Z1 comprises the amino acid sequence of SEQ ID NO:284. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:20 and Z1 comprises the amino acid sequence of SEQ ID NO:285.

[0147] In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 7, and Z1 comprises IL-22 or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 7, and Z1 comprises IL-22 or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 7, and Z1 comprises IL-22 or a fragment thereof. In some embodiments, Z1 comprises IL-22. In some embodiments, Z1 comprises a fragment of IL-22.

[0148] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:7, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:22, SEQ ID NO:59, or SEQ ID NO:282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:7, and Z1 comprises the amino acid sequence of SEQ ID NO:22, SEQ ID NO:59, or SEQ ID NO:282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO:7, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO:22, SEQ ID NO:59, or SEQ ID NO:282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO:7, and Z1 comprises the amino acid sequence of SEQ ID NO:22, SEQ ID NO:59, or SEQ ID NO:282, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO:22, SEQ ID NO:59, or SEQ ID NO:282, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7, and Z1 comprises the amino acid sequence of SEQ ID NO:22, SEQ ID NO:59, or SEQ ID NO:282, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7, and Z1 comprises the amino acid sequence of SEQ ID NO:22, or a fragment thereof.In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 7 and Z1 comprises the amino acid sequence of SEQ ID NO: 59, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 7 and Z1 comprises the amino acid sequence of SEQ ID NO: 282, or a fragment thereof.

[0149] In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 14, and Z1 comprises IL-22 or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 14, and Z1 comprises IL-22 or a fragment thereof. In some embodiments, Z1 comprises IL-22. In some embodiments, Z1 comprises a fragment of IL-22.

[0150] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 14, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 14, and Z1 comprises the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 14, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 14, and Z1 comprises the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14, and Z1 comprises the amino acids of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14, and Z1 comprises the amino acids of SEQ ID NO: 22, or a fragment thereof.In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14 and Z1 comprises the amino acid sequence of SEQ ID NO: 59, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14 and Z1 comprises the amino acid sequence of SEQ ID NO: 282, or a fragment thereof.

[0151] In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises IL-22 or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises IL-22 or a fragment thereof. In some embodiments, Z1 comprises IL-22. In some embodiments, Z1 comprises a fragment of IL-22.

[0152] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises amino acids of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 148, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 148, and Z1 comprises amino acids of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 148, and Z1 comprises amino acids of SEQ ID NO: 22, or a fragment thereof.In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 148 and Z1 comprises the amino acid sequence of SEQ ID NO: 59, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 148 and Z1 comprises the amino acid sequence of SEQ ID NO: 282, or a fragment thereof.

[0153] In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises IL-22 or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises IL-22 or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 180, and Z1 comprises IL-22 or a fragment thereof. In some embodiments, Z1 comprises IL-22. In some embodiments, Z1 comprises a fragment of IL-22.

[0154] In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 180, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 180, and Z1 comprises the amino acids of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 180, and Z1 comprises the amino acids of SEQ ID NO: 22, or a fragment thereof.In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 180 and Z1 comprises the amino acid sequence of SEQ ID NO: 59, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 180 and Z1 comprises the amino acid sequence of SEQ ID NO: 282, or a fragment thereof.

[0155] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO:6, and Z1 is a payload protein comprising an anti-TNFα antibody or an anti-TNFα antibody fragment. In some embodiments, Z1 comprises an amino acid sequence selected from SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:6, and Z1 comprises the amino acid sequence of SEQ ID NO:21. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:6, and Z1 comprises the amino acid sequence of SEQ ID NO:118. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:6, and Z1 comprises the amino acid sequence of SEQ ID NO:119. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:6, and Z1 comprises the amino acid sequence of SEQ ID NO:120.

[0156] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO:7, and Z1 is a payload protein comprising an anti-TNFα antibody or an anti-TNFα antibody fragment. In some embodiments, Z1 comprises an amino acid sequence selected from SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7, and Z1 comprises the amino acid sequence of SEQ ID NO:21. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7, and Z1 comprises the amino acid sequence of SEQ ID NO:118. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7, and Z1 comprises the amino acid sequence of SEQ ID NO:119. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7, and Z1 comprises the amino acid sequence of SEQ ID NO:120.

[0157] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 14, and Z1 is a payload protein comprising an anti-TNFα antibody or an anti-TNFα antibody fragment. In some embodiments, Z1 comprises an amino acid sequence selected from SEQ ID NO: 21, SEQ ID NO: 118, SEQ ID NO: 119, or SEQ ID NO: 120. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14, and Z1 comprises the amino acid sequence of SEQ ID NO: 21. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14, and Z1 comprises the amino acid sequence of SEQ ID NO: 118. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14, and Z1 comprises the amino acid sequence of SEQ ID NO: 119. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14, and Z1 comprises the amino acid sequence of SEQ ID NO: 120.

[0158] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO:6 and Z1 is a payload protein comprising leptin. In some embodiments, Z1 comprises the amino acid sequence of SEQ ID NO:25. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:6 and Z1 comprises the amino acid sequence of SEQ ID NO:25.

[0159] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO:7 and Z1 is a payload protein comprising leptin. In some embodiments, Z1 comprises the amino acid sequence of SEQ ID NO:25. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7 and Z1 comprises the amino acid sequence of SEQ ID NO:25.

[0160] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 14 and Z1 is a payload protein comprising leptin. In some embodiments, Z1 comprises the amino acid sequence of SEQ ID NO: 25. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14 and Z1 comprises the amino acid sequence of SEQ ID NO: 25.

[0161] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 17 and Z1 is a payload protein comprising leptin. In some embodiments, Z1 comprises the amino acid sequence of SEQ ID NO: 25. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 17 and Z1 comprises the amino acid sequence of SEQ ID NO: 25.

[0162] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 18 and Z1 is a payload protein comprising leptin. In some embodiments, Z1 comprises the amino acid sequence of SEQ ID NO: 25. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 18 and Z1 comprises the amino acid sequence of SEQ ID NO: 25.

[0163] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 19 and Z1 is a payload protein comprising leptin. In some embodiments, Z1 comprises the amino acid sequence of SEQ ID NO: 25. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 19 and Z1 comprises the amino acid sequence of SEQ ID NO: 25.

[0164] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO:47 and Z1 is a payload protein comprising leptin. In some embodiments, Z1 comprises the amino acid sequence of SEQ ID NO:25. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:47 and Z1 comprises the amino acid sequence of SEQ ID NO:25.

[0165] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO:48 and Z1 is a payload protein comprising leptin. In some embodiments, Z1 comprises the amino acid sequence of SEQ ID NO:25. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:48 and Z1 comprises the amino acid sequence of SEQ ID NO:25.

[0166] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 148 and Z1 is a payload protein comprising leptin. In some embodiments, Z1 comprises the amino acid sequence of SEQ ID NO: 25. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 148 and Z1 comprises the amino acid sequence of SEQ ID NO: 25.

[0167] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 180 and Z1 is a payload protein comprising leptin. In some embodiments, Z1 comprises the amino acid sequence of SEQ ID NO: 25. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 180 and Z1 comprises the amino acid sequence of SEQ ID NO: 25.

[0168] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO:20 and Z1 is a payload protein comprising IL-10. In some embodiments, Z1 comprises the amino acid sequence of SEQ ID NO:24, SEQ ID NO:283, SEQ ID NO:284, or SEQ ID NO:285. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:20 and Z1 comprises the amino acid sequence of SEQ ID NO:24. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:20 and Z1 comprises the amino acid sequence of SEQ ID NO:283. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:20 and Z1 comprises the amino acid sequence of SEQ ID NO:284. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:20 and Z1 comprises the amino acid sequence of SEQ ID NO:285.

[0169] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO:7 and Z1 is a payload protein comprising IL-22. In some embodiments, Z1 comprises the amino acid sequence of SEQ ID NO:22, SEQ ID NO:59, or SEQ ID NO:282. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7 and Z1 comprises the amino acid sequence of SEQ ID NO:22. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7 and Z1 comprises the amino acid sequence of SEQ ID NO:59. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7 and Z1 comprises the amino acid sequence of SEQ ID NO:282.

[0170] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 14, and Z1 is a payload protein comprising IL-22. In some embodiments, Z1 comprises the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14, and Z1 comprises the amino acid sequence of SEQ ID NO: 22. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14, and Z1 comprises the amino acid sequence of SEQ ID NO: 59. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14, and Z1 comprises the amino acid sequence of SEQ ID NO: 282.

[0171] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 148 and Z1 is a payload protein comprising IL-22. In some embodiments, Z1 comprises the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 148 and Z1 comprises the amino acid sequence of SEQ ID NO: 22. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 148 and Z1 comprises the amino acid sequence of SEQ ID NO: 59. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 148 and Z1 comprises the amino acid sequence of SEQ ID NO: 282.

[0172] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 180 and Z1 is a payload protein comprising IL-22. In some embodiments, Z1 comprises the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 180 and Z1 comprises the amino acid sequence of SEQ ID NO: 22. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 180 and Z1 comprises the amino acid sequence of SEQ ID NO: 59. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 180 and Z1 comprises the amino acid sequence of SEQ ID NO: 282.

[0173] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 144, and Z1 is a payload protein comprising IL-22. In some embodiments, IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 144, and Z1 comprises the amino acid sequence of SEQ ID NO: 22. In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 145, and Z1 is a payload protein comprising IL-22. In some embodiments, IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 145, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0174] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 146, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 146, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0175] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 147, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 147, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0176] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 149, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 149, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0177] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 150, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 150, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0178] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 151, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 151, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0179] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 152, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 152, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0180] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 153, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 153, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0181] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 154, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 154, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0182] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 155, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 155, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0183] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 156, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 156, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0184] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 157, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 157, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0185] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 158, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 158, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0186] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 159, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 159, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0187] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 160, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 160, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0188] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 161, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 161, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0189] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 162, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 162, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0190] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 163, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 163, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0191] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 164, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 164, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0192] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 165, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 165, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0193] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 166, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 166, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0194] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 167, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 167, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0195] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 168, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 168, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0196] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 169, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 169, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0197] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 170, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 170, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0198] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 171, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 171, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0199] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 172, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 172, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0200] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 173, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 173, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0201] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 174, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 174, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0202] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 175, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 175, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0203] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 176, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 176, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0204] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 177, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 177, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0205] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 178, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 178, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0206] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 179, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 179, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0207] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 181, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 181, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0208] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 182, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 182, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0209] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 183, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 183, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0210] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 184, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 184, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0211] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 185, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 185, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0212] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 186, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 186, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0213] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 187, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 187, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0214] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 188, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 188, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0215] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 189, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 189, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0216] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 190, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 190, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0217] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 191, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 191, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0218] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 192, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 192, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0219] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 193, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 193, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0220] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 194, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 194, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0221] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 195, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 195, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0222] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 196, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 196, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0223] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 197, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 197, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0224] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 198, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 198, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0225] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 199, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 199, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0226] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 200, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 200, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0227] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO:201, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:201, and Z1 comprises the amino acid sequence of SEQ ID NO:22.

[0228] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO:202, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:202, and Z1 comprises the amino acid sequence of SEQ ID NO:22.

[0229] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO:203, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:203, and Z1 comprises the amino acid sequence of SEQ ID NO:22.

[0230] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO:204, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:204, and Z1 comprises the amino acid sequence of SEQ ID NO:22.

[0231] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 205, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 205, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0232] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO:206, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:206, and Z1 comprises the amino acid sequence of SEQ ID NO:22.

[0233] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO:207, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:207, and Z1 comprises the amino acid sequence of SEQ ID NO:22.

[0234] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO:208, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:208, and Z1 comprises the amino acid sequence of SEQ ID NO:22.

[0235] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO:209, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:209, and Z1 comprises the amino acid sequence of SEQ ID NO:22.

[0236] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 210, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 210, and Z1 comprises the amino acid sequence of SEQ ID NO: 22.

[0237] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO:211, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:211, and Z1 comprises the amino acid sequence of SEQ ID NO:22.

[0238] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO:212, and Z1 is a payload protein comprising IL-22. In some embodiments, the IL-22 is as provided herein. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:212, and Z1 comprises the amino acid sequence of SEQ ID NO:22.

[0239] In some embodiments, X1 and Z1 are not linked by a linker polypeptide. In some embodiments, X1 and Z1 are linked by a linker polypeptide. In some embodiments, the linker polypeptide can be any suitable linker polypeptide. In some embodiments, the linker polypeptide is selected from the group consisting of a flexible peptide linker, a rigid peptide linker, a cleavable peptide linker, or a non-cleavable peptide linker. In some embodiments, the linker polypeptide is a flexible peptide linker. In some embodiments, the linker polypeptide is a rigid peptide linker. In some embodiments, the linker polypeptide is a cleavable peptide linker. In some embodiments, the linker polypeptide is a non-cleavable peptide linker. Non-limiting exemplary linker polypeptides are shown in Table 4 below. [Table 4]

[0240] nucleic acid molecule In some embodiments, a nucleic acid molecule is provided. In some embodiments, the nucleic acid molecule encodes a signal peptide provided herein. In some embodiments, the nucleic acid molecule encodes a polypeptide comprising the formula of X1-Z1 provided herein.

[0241] In some embodiments, nucleic acid molecules encoding signal peptides provided herein comprise nucleic acid sequences set forth in Table 2. It should be understood that the sequence identifiers listed above are exemplary only and are not limiting in any way. Due to the degenerate nature of codons, one of skill in the art will readily appreciate that alternative nucleic acid molecules may encode the same signal peptide. Thus, in some embodiments, nucleic acid molecules encoding signal peptides provided herein comprise sequences having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the nucleic acid sequences set forth in Table 2.In some embodiments, the nucleic acid molecules encoding the signal peptides provided herein are selected from the group consisting of SEQ ID NO:132, SEQ ID NO:133, SEQ ID NO:134, SEQ ID NO:135, SEQ ID NO:136, SEQ ID NO:137, SEQ ID NO:138, SEQ ID NO:139, SEQ ID NO:140, SEQ ID NO:213, SEQ ID NO:214, SEQ ID NO:215, SEQ ID NO:216, SEQ ID NO:217, SEQ ID NO:218, SEQ ID NO:219, SEQ ID NO:220, SEQ ID NO:221, SEQ ID NO:222, SEQ ID NO:223, SEQ ID NO:224, SEQ ID NO:225, SEQ ID NO:226, SEQ ID NO:227, SEQ ID NO:228, SEQ ID NO:229, SEQ ID NO:230, SEQ ID NO:231, SEQ ID NO:232, SEQ ID NO:233, SEQ ID NO:234, SEQ ID NO:235, SEQ ID NO:236, SEQ ID NO:237, SEQ ID NO:238, SEQ ID NO:239, SEQ ID NO:240, SEQ ID NO:241, SEQ ID NO:242, SEQ ID NO:243, SEQ ID NO:244, SEQ ID NO:245, SEQ ID NO:246, SEQ ID NO:247, SEQ ID NO:248, SEQ ID NO:249, SEQ ID NO:250, SEQ ID NO:251, SEQ ID NO:252, SEQ ID NO:253, SEQ ID NO:254, SEQ ID NO:255, SEQ ID NO:256, SEQ ID NO:257, SEQ ID The nucleic acid sequence of sequence number 241, SEQ ID NO:242, SEQ ID NO:243, SEQ ID NO:244, SEQ ID NO:245, SEQ ID NO:246, SEQ ID NO:247, SEQ ID NO:248, SEQ ID NO:249, SEQ ID NO:250, SEQ ID NO:251, SEQ ID NO:252, SEQ ID NO:253, SEQ ID NO:254, SEQ ID NO:255, SEQ ID NO:256, SEQ ID NO:257, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:261, SEQ ID NO:262, SEQ ID NO:263, SEQ ID NO:264, SEQ ID NO:265, SEQ ID NO:266, SEQ ID NO:267, SEQ ID NO:268, SEQ ID NO:269, SEQ ID NO:270, SEQ ID NO:271, SEQ ID NO:272, SEQ ID NO:273, SEQ ID NO:274, SEQ ID NO:275, SEQ ID NO:276, SEQ ID NO:277, SEQ ID NO:278, SEQ ID NO:279, SEQ ID NO:280, or SEQ ID NO:281.In some embodiments, the nucleic acid molecules encoding the signal peptides provided herein are selected from the group consisting of SEQ ID NO:132, SEQ ID NO:133, SEQ ID NO:134, SEQ ID NO:135, SEQ ID NO:136, SEQ ID NO:137, SEQ ID NO:138, SEQ ID NO:139, SEQ ID NO:140, SEQ ID NO:213, SEQ ID NO:214, SEQ ID NO:215, SEQ ID NO:216, SEQ ID NO:217, SEQ ID NO:218, SEQ ID NO:219, SEQ ID NO:220, SEQ ID NO:221, SEQ ID NO:222, SEQ ID NO:223, SEQ ID NO:224, SEQ ID NO:225, SEQ ID NO:226, SEQ ID NO:227, SEQ ID NO:228, SEQ ID NO:229, SEQ ID NO:230, SEQ ID NO:231, SEQ ID NO:232, SEQ ID NO:233, SEQ ID NO:234, SEQ ID NO:235, SEQ ID NO:236, SEQ ID NO:237, SEQ ID NO:238, SEQ ID NO:239, SEQ ID NO:240, SEQ ID NO:241, SEQ ID NO:242, SEQ ID NO:243, SEQ ID NO:244, SEQ ID NO:245, SEQ ID NO:246, SEQ ID NO:247, SEQ ID NO:248, SEQ ID NO:249, SEQ ID NO:250, SEQ ID NO:251, SEQ ID NO:252, SEQ ID NO:253, SEQ ID NO:254, SEQ ID NO:255, SEQ ID NO:256, SEQ ID NO:257, SEQ ID SEQ ID NO:248, SEQ ID NO:249, SEQ ID NO:250, SEQ ID NO:251, SEQ ID NO:252, SEQ ID NO:253, SEQ ID NO:254, SEQ ID NO:255, SEQ ID NO:256, SEQ ID NO:257, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:261, SEQ ID NO:262, SEQ ID NO:263, SEQ ID NO:264, SEQ ID NO:265, SEQ ID NO:266, SEQ ID NO:267, SEQ ID NO:268, SEQ ID NO:269, SEQ ID NO:270, SEQ ID NO:271, SEQ ID NO:272 2, comprising a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the nucleic acid sequence of SEQ ID NO:273, SEQ ID NO:274, SEQ ID NO:275, SEQ ID NO:276, SEQ ID NO:277, SEQ ID NO:278, SEQ ID NO:279, SEQ ID NO:280, or SEQ ID NO:281.

[0242] bacteria In some embodiments, a bacterium is provided. In some embodiments, the bacterium comprises a nucleic acid molecule provided herein. In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a payload protein provided herein. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in Table 1. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to an amino acid sequence set forth in Table 1. In some embodiments, X1 comprises an amino acid sequence set forth in Table 1. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:148, or SEQ ID NO:180. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to an amino acid sequence selected from SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:148, or SEQ ID NO:180. In some embodiments, X1 comprises an amino acid sequence selected from SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:148, or SEQ ID NO:180.

[0243] In some embodiments, the polypeptide comprising formula X1-Z1 is as described herein.

[0244] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 6. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 6. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 6.

[0245] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 7. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 7. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 7.

[0246] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 14. In some embodiments, X1 comprises an amino acid sequence at least 98% identical to the amino acid sequence of SEQ ID NO: 14. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14.

[0247] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 17. In some embodiments, X1 comprises an amino acid sequence at least 98% identical to the amino acid sequence of SEQ ID NO: 17. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 17.

[0248] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 18. In some embodiments, X1 comprises an amino acid sequence at least 98% identical to the amino acid sequence of SEQ ID NO: 18. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 18.

[0249] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 19. In some embodiments, X1 comprises an amino acid sequence at least 98% identical to the amino acid sequence of SEQ ID NO: 19. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 19.

[0250] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 20. In some embodiments, X1 comprises an amino acid sequence at least 98% identical to the amino acid sequence of SEQ ID NO: 20. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 20.

[0251] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 47. In some embodiments, X1 comprises an amino acid sequence at least 98% identical to the amino acid sequence of SEQ ID NO: 47. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 47.

[0252] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 48. In some embodiments, X1 comprises an amino acid sequence at least 98% identical to the amino acid sequence of SEQ ID NO: 48. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 48.

[0253] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 148. In some embodiments, X1 comprises an amino acid sequence at least 98% identical to the amino acid sequence of SEQ ID NO: 148. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 148.

[0254] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 180. In some embodiments, X1 comprises an amino acid sequence at least 98% identical to the amino acid sequence of SEQ ID NO: 180. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 180.

[0255] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where Z1 comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence set forth in Table 3. In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where Z1 comprises an amino acid sequence at least 98% identical to an amino acid sequence set forth in Table 3. In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where Z1 comprises an amino acid sequence set forth in Table 3.

[0256] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where Z1 is a therapeutic protein. In some embodiments, Z1 is a payload protein selected from the group consisting of an antimicrobial peptide, catestatin, CRAMP, hBD1, hBD3, β-catethritin, dermaseptin, IL-10, IL-22, ClbS, TIGIT, TFF1, TFF2, TFF3, an anti-CCL2 antibody, an anti-CCL2 antibody fragment, an anti-IL23 antibody, an anti-IL23 antibody fragment, an anti-TNFα antibody, an anti-TNFα antibody fragment, GLP2, IGF1, leptin, and casein. In some embodiments, Z1 is a payload protein selected from the group consisting of an anti-TNFα antibody, an anti-TNFα antibody fragment, IL-10, IL-22, hBD3, leptin, and β-casein. In some embodiments, Z1 is an anti-TNFα antibody. In some embodiments, Z1 is an anti-TNFα antibody fragment. In some embodiments, Z1 is IL-10. In some embodiments, Z1 is IL-22. In some embodiments, Z1 is hBD3. In some embodiments, Z1 is leptin. In some embodiments, Z1 is β-casein.

[0257] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is an antimicrobial peptide. In some embodiments, the antimicrobial peptide is any antimicrobial peptide. In some embodiments, the antimicrobial peptide is hBD1, hBD2, hBD3, b-catethritin, dermaseptin, HD5, Reg3a, or C5a. In some embodiments, the antimicrobial peptide comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:23, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, or SEQ ID NO:55. In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is an antimicrobial peptide comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:23, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, or SEQ ID NO:55.

[0258] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is an enzyme. In some embodiments, the enzyme is any enzyme. In some embodiments, the enzyme is Clbs, shP28GST, Clbs (Gilliamella apicola), Clbs (Snodgrassella alvi), phytase, SUC2, SUIS, AnPep, β-galactosidase (Lactobacillus), LACPL exo-α-1,4-glucosidase, β-galactosidase (yeast), ACE2, xylose isomerase (RainFlu), xylose isomerase (Lactobacillus), MaIL, apyrase, or DL-endopeptidase. In some embodiments, the enzyme comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:74, SEQ ID NO:75, SEQ ID NO:76, SEQ ID NO:77, or SEQ ID NO:78. In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is an enzyme comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:74, SEQ ID NO:75, SEQ ID NO:76, SEQ ID NO:77, or SEQ ID NO:78.

[0259] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a food protein. In some embodiments, the food protein is any food protein. In some embodiments, the food protein is β-casein, α-casein, an α-casein-derived peptide, Ara h1, or Ara h2. In some embodiments, the food protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:89, or SEQ ID NO:90. In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 is a preprotein signal peptide described herein and Z1 is a food protein comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:89, or SEQ ID NO:90.

[0260] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a growth factor. In some embodiments, the growth factor is any growth factor. In some embodiments, the growth factor is selected from IGF1, CTB-IGF1, or elafin. In some embodiments, the growth factor comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:91, SEQ ID NO:92, or SEQ ID NO:93. In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a growth factor comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:91, SEQ ID NO:92, or SEQ ID NO:93.

[0261] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is an immune receptor. In some embodiments, the immune receptor is any immune receptor. In some embodiments, the immune receptor is TIGIT. In some embodiments, the immune receptor comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 104. In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is an immune receptor comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 104.

[0262] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a mucolytic agent. In some embodiments, the mucolytic agent is any mucolytic agent. In some embodiments, the mucolytic agent is DNAse 1. In some embodiments, the mucolytic agent comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 105. In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a mucolytic agent comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 105.

[0263] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a peptide. In some embodiments, the peptide is any peptide. In some embodiments, the peptide is selected from adrenomedullin or urmin. In some embodiments, the peptide comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 121 or SEQ ID NO: 122. In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a peptide comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:121 or SEQ ID NO:122.

[0264] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a filament-binding protein. In some embodiments, the filament-binding protein is any filament-binding protein. In some embodiments, the filament-binding protein is filaggrin. In some embodiments, the filament-binding protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 123. In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a filament-associated protein comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 123.

[0265] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:14, and Z1 comprises a nanobody. In some embodiments, the nanobody can be any nanobody. In some embodiments, the nanobody binds to a target protein, in which case the target protein can be any protein. In some embodiments, the nanobody binds to a target protein, in which case the target protein is TNFα, CCL2, IL23, EGFR-III, GLP2, VEGFB, ILR6, MERS, SARS, BVD, norovirus, RSV, TcdA, TcdB, PCSK9, Pseudomonas toxin, or IgE. In some embodiments, the Nanobody comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% to an amino acid sequence selected from SEQ ID NO:21, SEQ ID NO:106, SEQ ID NO:107, SEQ ID NO:108, SEQ ID NO:109, SEQ ID NO:110, SEQ ID NO:111, SEQ ID NO:112, SEQ ID NO:113, SEQ ID NO:114, SEQ ID NO:115, SEQ ID NO:116, SEQ ID NO:117, SEQ ID NO:118, SEQ ID NO:119, SEQ ID NO:120, SEQ ID NO:126, SEQ ID NO:127, SEQ ID NO:128, SEQ ID NO:129, SEQ ID NO:130, or SEQ ID NO:131.In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:14, and Z1 is selected from SEQ ID NO:21, SEQ ID NO:106, SEQ ID NO:107, SEQ ID NO:108, SEQ ID NO:109, SEQ ID NO:110, SEQ ID NO:111, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:59, SEQ ID NO:59, SEQ ID NO:112, or having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% to an amino acid sequence selected from SEQ ID NO:113, SEQ ID NO:114, SEQ ID NO:115, SEQ ID NO:116, SEQ ID NO:117, SEQ ID NO:118, SEQ ID NO:119, SEQ ID NO:120, SEQ ID NO:126, SEQ ID NO:127, SEQ ID NO:128, SEQ ID NO:129, SEQ ID NO:130, or SEQ ID NO:131.

[0266] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 comprises an amino acid having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:47, or SEQ ID NO:48, and Z1 comprises a hormone. In some embodiments, the hormone is any hormone. In some embodiments, the hormone is GLP1, exenatide-4, catestatin, leptin, nesfatin, linaclotide, plecanatide, uroguanylin, PYY, guanylin, or LCRF. In some embodiments, the hormone comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:25, SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:97, SEQ ID NO:98, SEQ ID NO:98, SEQ ID NO:99, SEQ ID NO:100, SEQ ID NO:101, SEQ ID NO:102, or SEQ ID NO:103. In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:47, or SEQ ID NO:48, and Z1 comprises a hormone comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:25, SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:97, SEQ ID NO:98, SEQ ID NO:99, SEQ ID NO:100, SEQ ID NO:101, SEQ ID NO:102, or SEQ ID NO:103.

[0267] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:20, SEQ ID NO:148, or SEQ ID NO:180, and Z1 comprises a cytokine. In some embodiments, the cytokine is any cytokine. In some embodiments, the cytokine is IL-10, TFF1, TFF2, TFF3, IL-22, or CTB-IL-22. In some embodiments, the cytokine comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:282, SEQ ID NO:283, SEQ ID NO:284, or SEQ ID NO:285. In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:20, SEQ ID NO:148, or SEQ ID NO:180, and Z1 comprises a cytokine comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:282, SEQ ID NO:283, SEQ ID NO:284, or SEQ ID NO:285.

[0268] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 6, and Z1 comprises an anti-TNF-α antibody or an anti-TNF-α antibody fragment. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 6, and Z1 comprises an anti-TNF-α antibody or an anti-TNF-α antibody fragment. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 6, and Z1 comprises an anti-TNF-α antibody or an anti-TNF-α antibody fragment. In some embodiments, Z1 comprises an anti-TNF-α antibody. In some embodiments, Z1 comprises an anti-TNFα antibody fragment. In some embodiments, the anti-TNFα antibody is as provided herein. In some embodiments, the anti-TNFα antibody fragment is an anti-TNFα antibody fragment provided herein.

[0269] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:6, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:6, and Z1 comprises the amino acid sequence of SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO:6, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO:6, and Z1 comprises the amino acid sequence of SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:6, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof.In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:6 and Z1 comprises amino acids of SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:6 and Z1 comprises amino acids of SEQ ID NO:21, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:6 and Z1 comprises amino acids of SEQ ID NO:118, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:6 and Z1 comprises amino acids of SEQ ID NO:119, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:6 and Z1 comprises amino acids of SEQ ID NO:120, or a fragment thereof.

[0270] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:7, and Z1 comprises an anti-TNF-α antibody or an anti-TNF-α antibody fragment. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO:7, and Z1 comprises an anti-TNF-α antibody or an anti-TNF-α antibody fragment. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 comprises an amino acid sequence having the amino acid sequence of SEQ ID NO:7, and Z1 comprises an anti-TNF-α antibody or an anti-TNF-α antibody fragment. In some embodiments, Z1 comprises an anti-TNF-α antibody. In some embodiments, Z1 comprises an anti-TNFα antibody fragment. In some embodiments, the anti-TNFα antibody is as provided herein. In some embodiments, the anti-TNFα antibody fragment is an anti-TNFα antibody fragment provided herein.

[0271] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:7, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:7, and Z1 comprises the amino acid sequence of SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO:7, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO:7 and Z1 comprises the amino acid sequence of SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7 and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof.In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7 and Z1 comprises amino acids of SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7 and Z1 comprises amino acids of SEQ ID NO:21, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7 and Z1 comprises amino acids of SEQ ID NO:118, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7 and Z1 comprises amino acids of SEQ ID NO:119, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7 and Z1 comprises amino acids of SEQ ID NO:120, or a fragment thereof.

[0272] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 14, and Z1 comprises an anti-TNF-α antibody or an anti-TNF-α antibody fragment. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 14, and Z1 comprises an anti-TNF-α antibody or an anti-TNF-α antibody fragment. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 14, and Z1 comprises an anti-TNF-α antibody or an anti-TNF-α antibody fragment. In some embodiments, Z1 comprises an anti-TNFα antibody. In some embodiments, Z1 comprises an anti-TNFα antibody fragment. In some embodiments, the anti-TNFα antibody is as provided herein. In some embodiments, the anti-TNFα antibody fragment is an anti-TNFα antibody fragment provided herein.

[0273] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:14, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 14, and Z1 comprises the amino acid sequence of SEQ ID NO: 21, SEQ ID NO: 118, SEQ ID NO: 119, or SEQ ID NO: 120, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 14, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 21, SEQ ID NO: 118, SEQ ID NO: 119, or SEQ ID NO: 120, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence at least 98% identical to the amino acid sequence of SEQ ID NO: 14, and Z1 comprises the amino acid sequence of SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:14, and Z1 comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:120, or a fragment thereof.In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14 and Z1 comprises amino acids of SEQ ID NO: 21, SEQ ID NO: 118, SEQ ID NO: 119, or SEQ ID NO: 120, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14 and Z1 comprises amino acids of SEQ ID NO: 21, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14 and Z1 comprises amino acids of SEQ ID NO: 118, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14 and Z1 comprises amino acids of SEQ ID NO: 119, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14 and Z1 comprises amino acids of SEQ ID NO: 120, or a fragment thereof.

[0274] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 17, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 17, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 17, and Z1 comprises leptin, or a fragment thereof. In some embodiments, Z1 comprises leptin. In some embodiments, Z1 comprises a fragment of leptin.

[0275] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 17, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 17, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 17, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 17, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 17 and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 17 and Z1 comprises the amino acids of SEQ ID NO: 25, or a fragment thereof.

[0276] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 18, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 18, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 18, and Z1 comprises leptin, or a fragment thereof. In some embodiments, Z1 comprises leptin. In some embodiments, Z1 comprises a fragment of leptin.

[0277] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 18, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 18, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 18, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 18, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 18 and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 18 and Z1 comprises the amino acids of SEQ ID NO: 25, or a fragment thereof.

[0278] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 19, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 19, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 19, and Z1 comprises leptin, or a fragment thereof. In some embodiments, Z1 comprises leptin. In some embodiments, Z1 comprises a fragment of leptin.

[0279] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 19, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 19, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 19, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 19, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 19 and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 19 and Z1 comprises the amino acids of SEQ ID NO: 25, or a fragment thereof.

[0280] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 47, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 47, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 47, and Z1 comprises leptin, or a fragment thereof. In some embodiments, Z1 comprises leptin. In some embodiments, Z1 comprises a fragment of leptin.

[0281] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:47, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 47, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 47, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 47, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acids of SEQ ID NO:47 and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:47 and Z1 comprises the amino acids of SEQ ID NO:25, or a fragment thereof.

[0282] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 48, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 48, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 48, and Z1 comprises leptin, or a fragment thereof. In some embodiments, Z1 comprises leptin. In some embodiments, Z1 comprises a fragment of leptin.

[0283] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:48, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 48, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 48, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 48, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:48 and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:48 and Z1 comprises the amino acids of SEQ ID NO:25, or a fragment thereof.

[0284] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises leptin, or a fragment thereof. In some embodiments, Z1 comprises leptin. In some embodiments, Z1 comprises a fragment of leptin.

[0285] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 148 and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 148 and Z1 comprises the amino acids of SEQ ID NO: 25, or a fragment thereof.

[0286] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises leptin, or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 180, and Z1 comprises leptin, or a fragment thereof. In some embodiments, Z1 comprises leptin. In some embodiments, Z1 comprises a fragment of leptin.

[0287] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises the amino acid sequence of SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 180 and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 180 and Z1 comprises the amino acids of SEQ ID NO: 25, or a fragment thereof.

[0288] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 20, and Z1 comprises IL-10 or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 20, and Z1 comprises IL-10 or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 20, and Z1 comprises IL-10 or a fragment thereof. In some embodiments, Z1 comprises IL-10. In some embodiments, Z1 comprises a fragment of IL-10.

[0289] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:20, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:24, SEQ ID NO:283, SEQ ID NO:284, or SEQ ID NO:285, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:20, and Z1 comprises the amino acid sequence of SEQ ID NO:24, SEQ ID NO:283, SEQ ID NO:284, or SEQ ID NO:285, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO:20, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO:24, SEQ ID NO:283, SEQ ID NO:284, or SEQ ID NO:285, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence at least 98% identical to the amino acid sequence of SEQ ID NO:20, and Z1 comprises the amino acid sequence of SEQ ID NO:24, SEQ ID NO:283, SEQ ID NO:284, or SEQ ID NO:285, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:20, and Z1 comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:24, SEQ ID NO:283, SEQ ID NO:284, or SEQ ID NO:285, or a fragment thereof.In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:20 and Z1 comprises the amino acid sequence of SEQ ID NO:24, SEQ ID NO:283, SEQ ID NO:284, or SEQ ID NO:285, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:20 and Z1 comprises the amino acid sequence of SEQ ID NO:24. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:20 and Z1 comprises the amino acid sequence of SEQ ID NO:283. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:20 and Z1 comprises the amino acid sequence of SEQ ID NO:284. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:20 and Z1 comprises the amino acid sequence of SEQ ID NO:285.

[0290] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:7, and Z1 comprises IL-22 or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO:7, and Z1 comprises IL-22 or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having the amino acid sequence of SEQ ID NO:7, and Z1 comprises IL-22 or a fragment thereof. In some embodiments, Z1 comprises IL-22. In some embodiments, Z1 comprises a fragment of IL-22.

[0291] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:7, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:22, SEQ ID NO:59, or SEQ ID NO:282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO:7, and Z1 comprises the amino acid sequence of SEQ ID NO:22, SEQ ID NO:59, or SEQ ID NO:282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO:7, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO:22, SEQ ID NO:59, or SEQ ID NO:282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO:7, and Z1 comprises the amino acid sequence of SEQ ID NO:22, SEQ ID NO:59, or SEQ ID NO:282, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO:22, SEQ ID NO:59, or SEQ ID NO:282, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO:7, and Z1 comprises the amino acid sequence of SEQ ID NO:22, SEQ ID NO:59, or SEQ ID NO:282, or a fragment thereof.In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 7 and Z1 comprises the amino acid sequence of SEQ ID NO: 22. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 7 and Z1 comprises the amino acid sequence of SEQ ID NO: 59. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 7 and Z1 comprises the amino acid sequence of SEQ ID NO: 282.

[0292] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 14, and Z1 comprises IL-22 or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 14, and Z1 comprises IL-22 or a fragment thereof. In some embodiments, Z1 comprises IL-22. In some embodiments, Z1 comprises a fragment of IL-22.

[0293] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 14, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 14, and Z1 comprises the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 14, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 14, and Z1 comprises the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14, and Z1 comprises the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof.In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14 and Z1 comprises the amino acid sequence of SEQ ID NO: 22. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14 and Z1 comprises the amino acid sequence of SEQ ID NO: 59. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 14 and Z1 comprises the amino acid sequence of SEQ ID NO: 282.

[0294] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises IL-22 or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises IL-22 or a fragment thereof. In some embodiments, Z1 comprises IL-22. In some embodiments, Z1 comprises a fragment of IL-22.

[0295] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 148, and Z1 comprises the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 148, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 148, and Z1 comprises the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof.In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 148 and Z1 comprises the amino acid sequence of SEQ ID NO: 22. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 148 and Z1 comprises the amino acid sequence of SEQ ID NO: 59. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 148 and Z1 comprises the amino acid sequence of SEQ ID NO: 282.

[0296] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises IL-22 or a fragment thereof. In some embodiments, the polypeptide comprises the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises IL-22 or a fragment thereof. In some embodiments, Z1 comprises IL-22. In some embodiments, Z1 comprises a fragment of IL-22.

[0297] In some embodiments, the bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula of X1-Z1, where X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 180, and Z1 comprises the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 180, and Z1 comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 180, and Z1 comprises the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 59, or SEQ ID NO: 282, or a fragment thereof.In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 180 and Z1 comprises the amino acid sequence of SEQ ID NO: 22. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 180 and Z1 comprises the amino acid sequence of SEQ ID NO: 59. In some embodiments, X1 comprises the amino acid sequence of SEQ ID NO: 180 and Z1 comprises the amino acid sequence of SEQ ID NO: 282.

[0298] In some embodiments, the bacterium is a bacterium provided herein. In some embodiments, the bacterium is a Lactobacillus bacterium provided herein. In some embodiments, the Lactobacillus is Lactobacillus acetotolerans, Lactobacillus acidifarinae, Lactobacillus acidipiscis, Lactobacillus acidophilus, Lactobacillus agilis, Lactobacillus algidus, Lactobacillus alimentarius, Lactobacillus alvei, Lactobacillus alvi, Lactobacillus amylolyticus, Lactobacillus amylophilus, Lactobacillus amylotrophicus, Lactobacillus amylovorus, Lactobacillus animalis, Lactobacillus animata, Lactobacillus antri, Lactobacillus apinorum, Lactobacillus apis, Lactobacillus apodemi, Lactobacillus aquaticus, Lactobacillus aviarius, Lactobacillus backii, Lactobacillus bifermentans, Lactobacillus bombi, Lactobacillus bombicola, Lactobacillus brantae, Lactobacillus brevis, Lactobacillus brevisimilis, Lactobacillus buchneri, Lactobacillus cacaonum, Lactobacillus camelliae, Lactobacillus capillatus, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus zeae, Lactobacillus catenefornis, Lactobacillus ceti, Lactobacilluscoleohominis, Lactobacillus colini, Lactobacillus collinoides, Lactobacillus composti, Lactobacillus concavus, Lactobacillus coryniformis, Lactobacillus crispatus, Lactobacillus crustorum, Lactobacillus curieae, Lactobacillus curvatus, Lactobacillus delbrueckii, Lactobacillus dextrinicus, Lactobacillus diolivorans, Lactobacillus equi, Lactobacillus equicursoris, Lactobacillus equigenerosi, Lactobacillus fabifermentans, Lactobacillus faecis, Lactobacillus faeni, Lactobacillus farciminis, Lactobacillus farraginis, Lactobacillus fermentum, Lactobacillus floricola, Lactobacillus florum, Lactobacillus formosensis, Lactobacillus fornicalis, Lactobacillus fructivorans, Lactobacillus frumenti, Lactobacillus fuchuensis, Lactobacillus furfuricola, Lactobacillus futsaii, Lactobacillus gallinarum, Lactobacillus gasseri, Lactobacillus gastricus, Lactobacillus ghanensis, Lactobacillus gigeriorum, Lactobacillus ginsenosidimutans, Lactobacillus gorillae, Lactobacillus graminis, Lactobacillus guizhouensis, Lactobacillushalophilus, Lactobacillus hammesii, Lactobacillus hamsteri, Lactobacillus harbinensis, Lactobacillus hayakitensis, Lactobacillus heilongjiangensis, Lactobacillus helsingborgensis, Lactobacillus helveticus, Lactobacillus herbarum, Lactobacillus heterohiochii, Lactobacillus hilgardii, Lactobacillus hokkaidonensis, Lactobacillus hominis, Lactobacillus homohiochii, Lactobacillus hordei, Lactobacillus iatae, Lactobacillus iners, Lactobacillus ingluviei, Lactobacillus insectis, Lactobacillus insicii, Lactobacillus intermedius, Lactobacillus intestinalis, Lactobacillus iwatensis, Lactobacillus ixorae, Lactobacillus japonicus, Lactobacillus jensenii, Lactobacillus johnsonii, Lactobacillus kalixensis, Lactobacillus kefiranofaciens, Lactobacillus kefiri, Lactobacillus kimbladii, Lactobacillus kimchicus, Lactobacillus kimchiensis, Lactobacillus kisonensis, Lactobacillus kitasatonis, Lactobacillus koreensis, Lactobacillus kullabergensis、Lactobacillus kunkeei、Lactobacillus larvae、Lactobacillus leichmannii、Lactobacillusletivazi, Lactobacillus lindneri, Lactobacillus malefermentans, Lactobacillus mali, Lactobacillus manihotivorans, Lactobacillus mellifer, Lactobacillus mellis, Lactobacillus melliventris, Lactobacillus micheneri, Lactobacillus mindensis, Lactobacillus mixtipabuli, Lactobacillus mobilis, Lactobacillus modestisalitolerans, Lactobacillus mucosae, Lactobacillus mudanjiangensis, Lactobacillus murinus, Lactobacillus nagelii, Lactobacillus namurensis, Lactobacillus nantensis, Lactobacillus nasuensis, Lactobacillus nenjiangensis, Lactobacillus nodensis, Lactobacillus odoratitofui, Lactobacillus oeni, Lactobacillus oligofermentans, Lactobacillus ori, Lactobacillus oryzae, Lactobacillus otakiensis, Lactobacillus ozensis, Lactobacillus panis, Lactobacillus pantheris, Lactobacillus parabrevis, Lactobacillus parabuchneri, Lactobacillus paracollinoides, Lactobacillus parafarraginis, Lactobacillus parakefiri, Lactobacillus paraalimentarius, Lactobacillus paraplantarum, Lactobacillus pasteurii, Lactobacillus paucivorans, Lactobacilluspentosus, Lactobacillus perolens, Lactobacillus plajomi, Lactobacillus plantarum, Lactobacillus pobuzihii, Lactobacillus pontis, Lactobacillus porcinae, Lactobacillus psittaci, Lactobacillus rapi, Lactobacillus rennanquilfy, Lactobacillus rennini, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus rodentium, Lactobacillus rogosae, Lactobacillus rossiae, Lactobacillus ruminis, Lactobacillus saerimneri, Lactobacillus sakei, Lactobacillus salivarius, Lactobacillus sanfranciscensis, Lactobacillus saniviri, Lactobacillus satsumensis, Lactobacillus secaliphilus, Lactobacillus selangorensis, Lactobacillus senioris, Lactobacillus senmaizukei, Lactobacillus sharpeae, Lactobacillus shenzhenensis, Lactobacillus sicerae, Lactobacillus silagei, Lactobacillus siliginis, Lactobacillus similis, Lactobacillus songhuajiangensis, Lactobacillus spicheri, Lactobacillus sucicola, Lactobacillus suebicus、Lactobacillus sunkii、Lactobacillus taiwanensis、Lactobacillus thailandensis、Lactobacillus tucceti、Lactobacillusultunensis, Lactobacillus uvarum, Lactobacillus vaccinostercus, Lactobacillus vaginalis, Lactobacillus vermiforme, Lactobacillus vespulae, Lactobacillus vini, Lactobacillus wasatchensis, Lactobacillus xiang fangensis, Lactobacillus yonginensis, Lactobacillus zymae, Atopobium (e.g. A. deltae), Carnobacterium (e.g. C. alterfunditum), Weissella (e.g. W. beninensis), Oenococcus (e.g. O. oeni), L euconostoc (e.g. L. carnosum), Pediococcus (e.g. P. acidilactici), Paralactobacillus (e.g. P. selangorensis), Holzapfelia (e.g. H. fl oricola), Amylolactobacillus (e.g., A. amylophilus), Bombilactobacillus (e.g., B. mellifer), Companilactobacillus (e.g., C. alimentarius), Lapidilactobacillus (e.g., L. concavus), Agrilactobacillus (e.g., A. composti), Schleiferilactobacillus (e.g., S. perolens), Loi golactobacillus (e.g., L. coryniformis), Lacticaseibacillus (e.g., L. casei), Latilactobacillus (e.g., L. sakei), Dellaglioa (e.g., D. algidus), Liquorilactobacillus (e.g., L. mail), Ligilactobacillus (e.g., L. salivarius), Lactiplantibacillus (e.g., L. plantarum), Furfurilactobacillus (e.g., F. rossiae), Paucilactobacillus (e.g., P. vaccinostercus), Limosilactobacillus (e.g., L. fermentum), Fructilactobacillus (e.g., F. fructivorans), Acetilactobacillus (e.g., A. jinshani), Apilactobacillus (e.g., A. kunkeei), Levilactobacillus (L.brevis), Secundilactobacillus (e.g., S. collinoides), or Lentilactobacillus (e.g., L. buchneri). In some embodiments, the bacterium is a Lactobacillus bacterium selected from L. gasseri, L. jensenii, L. crispatus, L. rhamnosus, L. casei, L. paracasei, L. acidophilus, L. delbruckii, L. johnsonii, L. fermentum, L. reuteri, and L. salivarus. In some embodiments, the Lactobacillus bacterium is L. gasseri. In some embodiments, the Lactobacillus bacterium is L. jensenii. In some embodiments, the Lactobacillus bacterium is L. crispatus. In some embodiments, the Lactobacillus bacterium is L. rhamnosus. In some embodiments, the Lactobacillus bacterium is L. casei. In some embodiments, the Lactobacillus bacterium is L. paracasei. In some embodiments, the Lactobacillus bacterium is L. acidophilus. In some embodiments, the Lactobacillus bacterium is L. delbruckii. In some embodiments, the Lactobacillus bacterium is L. johnsonii. In some embodiments, the Lactobacillus bacterium is L. fermentum. In some embodiments, the Lactobacillus bacterium is L. reuteri. In some embodiments, the Lactobacillus bacterium is L. salivarus. In some embodiments, the Lactobacillus bacterium is selected from L. gasseri and L. rhamnosus. In some embodiments, the Lactobacillus bacterium is L. gasseri. In some embodiments, the Lactobacillus bacterium is L. rhamnosus.

[0299] In some embodiments, an L. gasseri bacterium is provided. In some embodiments, the L. gasseri bacterium comprises a nucleic acid molecule provided herein. In some embodiments, the L. gasseri bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a payload protein provided herein.

[0300] In some embodiments, an L. jensenii bacterium is provided. In some embodiments, the L. jensenii bacterium comprises a nucleic acid molecule provided herein. In some embodiments, the L. jensenii bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a payload protein provided herein.

[0301] In some embodiments, an L. crispatus bacterium is provided. In some embodiments, the L. crispatus bacterium comprises a nucleic acid molecule provided herein. In some embodiments, the L. crispatus bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a payload protein provided herein.

[0302] In some embodiments, an L. rhamnosus bacterium is provided. In some embodiments, the L. rhamnosus bacterium comprises a nucleic acid molecule provided herein. In some embodiments, the L. rhamnosus bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a payload protein provided herein.

[0303] In some embodiments, an L. casei bacterium is provided. In some embodiments, the L. casei bacterium comprises a nucleic acid molecule provided herein. In some embodiments, the L. casei bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a payload protein provided herein.

[0304] In some embodiments, L. paracasei bacteria are provided. In some embodiments, the L. paracasei bacteria comprise a nucleic acid molecule provided herein. In some embodiments, the L. paracasei bacteria comprise a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a payload protein provided herein.

[0305] In some embodiments, an L. acidophilus bacterium is provided. In some embodiments, the L. acidophilus bacterium comprises a nucleic acid molecule provided herein. In some embodiments, the L. acidophilus bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a payload protein provided herein.

[0306] In some embodiments, L. delbruckii bacteria are provided. In some embodiments, the L. delbruckii bacteria comprise a nucleic acid molecule provided herein. In some embodiments, the L. delbruckii bacteria comprise a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a payload protein provided herein.

[0307] In some embodiments, an L. johnsonii bacterium is provided. In some embodiments, the L. johnsonii bacterium comprises a nucleic acid molecule provided herein. In some embodiments, the L. johnsonii bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a payload protein provided herein.

[0308] In some embodiments, an L. fermentum bacterium is provided. In some embodiments, the L. fermentum bacterium comprises a nucleic acid molecule provided herein. In some embodiments, the L. fermentum bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a payload protein provided herein.

[0309] In some embodiments, an L. reuteri bacterium is provided. In some embodiments, the L. reuteri bacterium comprises a nucleic acid molecule provided herein. In some embodiments, the L. reuteri bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a payload protein provided herein.

[0310] In some embodiments, an L. salivarus bacterium is provided. In some embodiments, the L. salivarus bacterium comprises a nucleic acid molecule provided herein. In some embodiments, the L. salivarus bacterium comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a payload protein provided herein.

[0311] composition In another aspect, the present embodiments provide compositions, e.g., pharmaceutically acceptable compositions, which comprise the modified microorganisms provided herein, which can be formulated, for example, with a pharmaceutically acceptable carrier. As used herein, "pharmaceutically acceptable carrier" includes all physiologically compatible solvents, dispersion media, isotonic and absorption delaying agents, and the like. The carrier can be suitable for intravenous, intramuscular, subcutaneous, parenteral, rectal, local, topical, spinal, or epidermal administration (e.g., by injection or inhalation).

[0312] In any of the embodiments provided herein, the pharmaceutical composition or composition comprises an engineered microorganism provided herein. In some embodiments, the engineered microorganism is any engineered microorganism provided herein. In some embodiments, the engineered microorganism is a Lactobacillus species provided herein. In some embodiments, the Lactobacillus is selected from L. gasseri, L. jensenii, L. crispatus, L. rhamnosus, L. casei, L. paracasei, L. acidophilus, L. delbruckii, L. johnsonii, L. fermentum, L. reuteri, and L. salivarus. In some embodiments, the Lactobacillus bacterium is L. gasseri. In some embodiments, the Lactobacillus bacterium is L. jensenii. In some embodiments, the Lactobacillus bacterium is L. crispatus. In some embodiments, the Lactobacillus bacterium is L. rhamnosus. In some embodiments, the Lactobacillus bacterium is L. casei. In some embodiments, the Lactobacillus bacterium is L. paracasei. In some embodiments, the Lactobacillus bacterium is L. acidophilus. In some embodiments, the Lactobacillus bacterium is L. delbruckii. In some embodiments, the Lactobacillus bacterium is L. johnsonii. In some embodiments, the Lactobacillus bacterium is L. fermentum. In some embodiments, the Lactobacillus bacterium is L. reuteri. In some embodiments, the Lactobacillus bacterium is L. salivarus. In some embodiments, the Lactobacillus bacterium is selected from L. gasseri and L. rhamnosus. In some embodiments, the Lactobacillus bacterium is L. gasseri. In some embodiments, the Lactobacillus bacterium is L. rhamnosus.

[0313] In some embodiments, the Lactobacillus bacterium of the composition comprises a nucleic acid molecule provided herein. In some embodiments, the Lactobacillus bacterium of the composition comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a payload protein provided herein. In some embodiments, the L. gasseri bacterium of the composition comprises a nucleic acid molecule provided herein. In some embodiments, the L. gasseri bacterium of the composition comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a payload protein provided herein. In some embodiments, the L. rhamnosus bacterium of the composition comprises a nucleic acid molecule provided herein. In some embodiments, the L. rhamnosus bacterium of the composition comprises a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a payload protein provided herein. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:148, or SEQ ID NO:180, provided herein. In some embodiments, Z1 is a payload protein selected from the group consisting of an antimicrobial peptide provided herein, catestatin, CRAMP, hBD1, hBD3, β-catethritin, dermaseptin, IL-10, IL-22, ClbS, TIGIT, TFF1, TFF2, TFF3, an anti-CCL2 antibody, an anti-CCL2 antibody fragment, an anti-IL23 antibody, an anti-IL23 antibody fragment, an anti-TNFα antibody, an anti-TNFα antibody fragment, GLP2, IGF1, leptin, and casein.In some embodiments, Z1 is a payload protein selected from the group consisting of an anti-TNFα antibody, an anti-TNFα antibody fragment, IL-10, IL-22, hBD3, leptin, and beta-casein. In some embodiments, Z1 is an anti-TNFα antibody. In some embodiments, Z1 is an anti-TNFα antibody fragment. In some embodiments, Z1 is IL-10. In some embodiments, Z1 is IL-22. In some embodiments, Z1 is hBD3. In some embodiments, Z1 is leptin. In some embodiments, Z1 is beta-casein.

[0314] In some embodiments, the composition further comprises one or more pharmaceutically acceptable carriers or excipients, including, but not limited to, carriers, solvents, cosolvents, emulsifiers, lubricants, disintegrants, binders, fillers, glidants, rheological agents, solubilizers, antibacterial agents, antioxidants, preservatives, colorants, flavoring agents, emollients, pH adjusters, etc. Certain carriers, solvents, co-solvents, emulsifiers, lubricants, disintegrants, binders, fillers, glidants, rheological agents, solubilizers, antimicrobial agents, antioxidants, preservatives, colorants, flavoring agents, emollients, pH adjusters, and the like are well known in the art, and any such particular carriers, solvents, co-solvents, emulsifiers, lubricants, disintegrants, binders, fillers, glidants, rheological agents, solubilizers, antimicrobial agents, antioxidants, preservatives, colorants, flavoring agents, emollients, pH adjusters, and the like are within the scope of the present disclosure.

[0315] Pharmaceutically acceptable carriers useful in the present disclosure are conventionally known in the art. The nature of the carrier can vary depending on the particular mode of administration employed. For example, oral administration typically includes a pharmaceutically and physiologically acceptable fluid such as water, physiological saline, balanced salt solution, aqueous dextrose, glycerol, or the like as a vehicle. In addition to a biologically neutral carrier, oral compositions may also contain auxiliary substances such as wetting or emulsifying agents, preservatives, and pH buffering agents. Exemplary pH-adjusting agents include acids (e.g., acetic acid, adipic acid, carbonic acid, citric acid, fumaric acid, phosphoric acid, sorbic acid, succinic acid, tartaric acid), bases (e.g., magnesium oxide, tribasic potassium phosphate), and pharmaceutically acceptable salts thereof.

[0316] In some embodiments, the composition may further comprise one or more cosmetically acceptable excipients. The cosmetically acceptable excipients may be in addition to or in place of the pharmaceutically acceptable carriers or excipients. In some embodiments, the cosmetically acceptable excipients are selected from the group consisting of preservatives, antioxidants, chelating agents, fillers, film-forming agents, fragrances, pH adjusters, and combinations thereof. In some embodiments, the cosmetically acceptable excipient is selected from the group consisting of hydroxyacetophenone, ethylhexylglycerin, sodium phytate, tromethamine, lactic acid, disodium phosphate, phenoxyethanol, xylose, HDI / trimethylol hexyllactone crosspolymer, lactose, ethylhexylglycerin, disodium EDTA, inorganic salts, glucose, caprylyl glycol, silica, hexylene glycol, sodium benzoate, fragrance / parfum, BHT, hydrated silica, dimethyl silicate, sodium citrate, citric acid, lactic acid, sodium hydroxide, VP / polycarbamyl polyglycol esters, hydrolyzed sesame protein PG-propylmethylsilanediol, glucose, silica, tocopherol, limonene, linalool, isohexadecane, and combinations thereof. In some embodiments, the cosmetically acceptable excipient contains a plurality of excipients selected from hydroxyacetophenone, ethylhexylglycerin, sodium phytate, tromethamine, lactic acid, disodium phosphate, phenoxyethanol, xylose, HDI / trimethylol hexyllactone crosspolymer, lactose, ethylhexylglycerin, disodium EDTA, inorganic salts, glucose, caprylyl glycol, silica, hexylene glycol, sodium benzoate, fragrance / parfum, BHT, hydrated silica, dimethyl silicate, sodium citrate, citric acid, lactic acid, sodium hydroxide, VP / polycarbamyl polyglycol esters, hydrolyzed sesame protein PG-propylmethylsilanediol, glucose, silica, tocopherol, limonene, linalool, or isohexadecane.

[0317] The compositions may be in a variety of forms. These include, for example, liquid, semi-solid, and solid dosage forms, such as liquid solutions (e.g., injectable and infusible solutions), dispersions or suspensions, liposomes, and suppositories. The forms vary depending on the mode of administration and therapeutic application. Typical compositions are in the form of an injectable or infusible solution. In one embodiment, the mode of administration is parenteral (e.g., intravenous, subcutaneous, intraperitoneal, intramuscular). In one embodiment, the composition is administered by intravenous infusion or injection. In some embodiments, the composition is administered by intramuscular or subcutaneous injection. In some embodiments, the composition is administered locally to a target site, for example, by injection or topical application. For example, the pharmaceutical composition can be lyophilized prior to administration to a patient and reconstituted for use.

[0318] As used herein, the phrases "parenteral administration" and "administered parenterally" refer to modes of administration other than enteral administration and topical administration, usually by injection, and include, but are not limited to, intravenous, intramuscular, intra-arterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intra-articular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion.

[0319] Therapeutic compositions should generally be sterile and stable under the conditions of manufacture and storage. The compositions can be formulated as solutions, microemulsions, dispersions, liposomes, or other ordered structures suitable for high concentration of the therapeutic molecule. Sterile injectable solutions can be prepared by incorporating the required amount of the active compound (i.e., therapeutic molecule) in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle containing a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, preparation methods can be vacuum drying and freeze-drying, which yield a powder of the active ingredient plus any additional desired ingredients from a previously sterile-filtered solution thereof. The proper fluidity of the solution can be maintained, for example, by the use of a coating such as lecithin, by maintaining the required particle size in the case of dispersions, and by the use of surfactants. Prolonged absorption of injectable compositions can be achieved by including in the composition an agent that delays absorption, for example, monostearate salts and gelatin.

[0320] As will be appreciated by those skilled in the art, the route and / or mode of administration will vary depending on the desired results. In certain embodiments, the modified microorganisms may be prepared with a carrier that will protect the modified microorganisms against rapid release, for example, a controlled-release formulation, including implants, transdermal patches, and microencapsulated delivery systems. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used. Many methods for the preparation of such formulations are patented or known to those skilled in the art. See, for example, *Sustained and Controlled Release Drug Delivery Systems*, *J.R. Robinson*, ed., *Marcel Dekker*, Inc., New York, 1978.

[0321] In some embodiments, the pharmaceutical composition can be administered orally, for example, with an inert diluent or an assimilable edible carrier. The compound (and other ingredients, if desired) can also be enclosed in hard or soft shell gelatin capsules, compressed into tablets, or incorporated directly into the subject's diet. For oral therapeutic administration, the compound can be mixed with excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. To administer the composition by a method other than parenteral administration, it may be necessary to coat the composition with, or administer the compound together with, a material that prevents its inactivation. Therapeutic compositions can also be administered using medical devices known in the art.

[0322] The dosage regimen is adjusted to provide the optimum desired response (e.g., therapeutic response). For example, a single bolus may be administered, multiple divided doses may be administered over time, or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. For ease of administration and uniformity of dose, it is particularly advantageous to formulate parenteral compositions in unit dosage form. As used herein, unit dosage form refers to physically discrete units suitable as dosage units for treating a subject, each unit containing a predetermined amount of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specifications for the unit dosage form are dictated by and directly depend on (a) the unique characteristics of the active compound and the specific therapeutic effect to be achieved, and (b) the inherent limitations of the technical field of formulating such active compounds for sensitive treatment in individuals.

[0323] A therapeutically effective amount of the modified microorganism may be measured or determined in colony forming units (CFU), and may be any amount, for example, about 100 CFU to 10 CFU, about 10 to 10 CFU, 10 to 10 CFU, or about 10 to about 10 CFU. In some embodiments, a therapeutically effective amount of the modified microorganism is about 100 CFU to about 10 CFU. In some embodiments, a therapeutically effective amount of the modified microorganism is about 100 CFU, about 10 CFU, or about 10 to about 10 CFU, about 10 CFU, about 10 CFU, or about 10 CFU. In some embodiments, a therapeutically effective amount of the modified microorganism is any CFU amount falling within any of the above ranges. Note that dosage values ​​may vary depending on the type and severity of the condition to be alleviated. It is further understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions, and that dosage ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed compositions.

[0324] A pharmaceutical composition may contain a "therapeutically effective amount" or a "prophylactically effective amount" of the composition or modified microorganism. A "therapeutically effective amount" refers to an amount effective, at a dosage and for a period of time necessary, to achieve a desired therapeutic result. The therapeutically effective amount of a therapeutic molecule may vary depending on factors such as the individual's disease state, age, and weight, as well as the ability of the modified microorganism to elicit a desired response in the individual. A therapeutically effective amount also refers to an amount in which the therapeutically beneficial effects outweigh the toxic or harmful effects of the modified microorganism. A "therapeutically effective amount" preferably reduces a measurable parameter, such as the content of microorganisms that cause vaginal dysbiosis, by at least about 20%, at least about 40%, at least about 60%, and at least about 80% compared to untreated subjects. The ability of a composition or modified microorganism to reduce a measurable parameter, such as the content of microorganisms that cause vaginal dysbiosis, can be evaluated in an animal model system that is predictive of efficacy in reducing the content of microorganisms that cause vaginal dysbiosis. Alternatively, this property of the composition or modified microorganism can be assessed by examining the ability of the composition or modified microorganism to inhibit the growth of microorganisms that cause vaginal dysbiosis in vitro, such in vitro inhibition being achieved by assays known to those skilled in the art.

[0325] A "prophylactically effective amount" refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result. Because a prophylactic dose is used in subjects prior to or at an earlier stage of disease, generally, the prophylactically effective amount can be less than the therapeutically effective amount.

[0326] Also provided herein are kits containing the compositions described herein, which may include instructions for use, other reagents (e.g., labels, therapeutic agents, or agents useful for chelating or attaching therapeutic molecules to labels or other therapeutic agents, or radioprotective compositions), devices or other materials for preparing the therapeutic molecules for administration, a pharmaceutically acceptable carrier, and devices or other materials for administration to a subject.

[0327] method In some embodiments, a method for producing a protein is provided. The protein produced can be any protein. In some embodiments, the protein is useful for treating a disease or disorder. In some embodiments, the protein is a therapeutic protein. In some embodiments, the method includes transfecting a bacterium with a nucleic acid molecule provided herein to produce a bacterium containing the nucleic acid molecule, and culturing the bacterium containing the nucleic acid molecule under conditions sufficient to grow the bacterium, wherein the bacterium expresses and secretes the payload protein.

[0328] In some embodiments, the nucleic acid molecule encodes a polypeptide provided herein. In some embodiments, the nucleic acid molecule encodes a polypeptide comprising the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a payload protein provided herein. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:148, or SEQ ID NO:180, provided herein. In some embodiments, Z1 is a payload protein provided herein. In some embodiments, Z1 is a payload protein selected from the group consisting of an antimicrobial peptide provided herein, catestatin, CRAMP, hBD1, hBD3, β-catethritin, dermaseptin, IL-10, IL-22, ClbS, TIGIT, TFF1, TFF2, TFF3, an anti-CCL2 antibody, an anti-CCL2 antibody fragment, an anti-IL23 antibody, an anti-IL23 antibody fragment, an anti-TNFα antibody, an anti-TNFα antibody fragment, GLP2, IGF1, leptin, and casein. In some embodiments, Z1 is a payload protein selected from the group consisting of an anti-TNFα antibody, an anti-TNFα antibody fragment, IL-10, IL-22, hBD3, leptin, and β-casein. In some embodiments, Z1 is an anti-TNFα antibody. In some embodiments, Z1 is an anti-TNFα antibody fragment. In some embodiments, Z1 is IL-10. In some embodiments, Z1 is IL-22. In some embodiments, Z1 is hBD3. In some embodiments, Z1 is leptin. In some embodiments, Z1 is β-casein.

[0329] In some embodiments, a method for producing a protein comprises transfecting a bacterium with a nucleic acid molecule encoding a polypeptide provided herein to produce a bacterium comprising the nucleic acid molecule, and culturing the bacterium comprising the nucleic acid molecule under conditions sufficient to grow the bacterium, wherein the bacterium expresses and secretes the payload protein. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a payload protein provided herein.

[0330] In some embodiments, a method for producing a protein comprises transfecting a bacterium with a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a pre-protein signal peptide provided herein and Z1 is a payload protein provided herein, to produce a bacterium comprising the nucleic acid molecule, and culturing the bacterium comprising the nucleic acid molecule under conditions sufficient to grow the bacterium, wherein the bacterium expresses and secretes the payload protein.

[0331] In some embodiments, the bacterium is a bacterium provided herein. In some embodiments, the bacterium is a Lactobacillus bacterium provided herein. In some embodiments, a method for producing a therapeutic protein includes transfecting a Lactobacillus with a nucleic acid molecule encoding a polypeptide comprising the formula X1-Z1, where X1 is a pre-protein signal peptide provided herein and Z1 is a payload protein provided herein, to produce a Lactobacillus comprising the nucleic acid molecule, and culturing the Lactobacillus comprising the nucleic acid molecule under conditions sufficient to grow the Lactobacillus, wherein the Lactobacillus expresses and secretes the payload protein. In some embodiments, the Lactobacillus bacterium is selected from L. gasseri, L. jensenii, L. crispatus, L. rhamnosus, L. casei, L. paracasei, L. acidophilus, L. delbruckii, L. johnsonii, L. fermentum, L. reuteri, and L. salivarus. In some embodiments, the Lactobacillus bacterium is L. gasseri. In some embodiments, the Lactobacillus bacterium is L. jensenii. In some embodiments, the Lactobacillus bacterium is L. crispatus. In some embodiments, the Lactobacillus bacterium is L. rhamnosus. In some embodiments, the Lactobacillus bacterium is L. casei. In some embodiments, the Lactobacillus bacterium is L. paracasei. In some embodiments, the Lactobacillus bacterium is L. acidophilus. In some embodiments, the Lactobacillus bacterium is L. delbruckii. In some embodiments, the Lactobacillus bacterium is L. johnsonii. In some embodiments, the Lactobacillus bacterium is L. fermentum. In some embodiments, the Lactobacillus bacterium is L. reuteri.In some embodiments, the Lactobacillus bacterium is L. salivarus. In some embodiments, the Lactobacillus bacterium is selected from L. gasseri and L. rhamnosus. In some embodiments, the Lactobacillus bacterium is L. gasseri. In some embodiments, the Lactobacillus bacterium is L. rhamnosus.

[0332] In some embodiments, the bacteria may be cultured for a period of time in an environment effective to maintain bacterial health, thereby producing a desired amount of therapeutic protein. Culturing bacteria is common practice and well known in the art. Generally, bacteria may be grown in a nutrient-rich broth, which may include amino acids and nitrogen. The engineered bacteria may be grown for any time necessary to produce a desired amount of recombinant payload protein, including a signal peptide and payload protein. For example, the bacteria may be grown for about 0.5 hours to about 168 hours or more. In some embodiments, the bacteria may be grown for 0.5 hours, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 18 hours, 24 hours, 30 hours, 36 hours, 42 hours, 48 ​​hours, 72 hours, 96 hours, 120 hours, 144 hours, or 168 hours or more. In some embodiments, the bacteria may be grown for any period of time falling within any of the recited periods or longer. Additionally, bacteria may be grown in a continuous culture system, whereby a portion of the bacterial culture is inoculated into fresh growth broth and the culture is continued. Thus, in some embodiments, bacteria may be grown for at least 0.5 hours. Those skilled in the art will recognize that growth time is a temperature-dependent variable, as different temperatures produce different bacterial growth rates, and thus, bacterial growth for any period of time is within the scope of the present application. Thus, the modified bacteria may be grown at room temperature, or more efficiently, at temperatures between about 40°C and 140°C, although any particular species and / or strain will have an optimal temperature range known to those skilled in the art. Temperature can be used to control bacterial growth and the production of the desired payload protein. Thus, in some embodiments, bacteria may be cultured at temperatures between about 4°C and about 140°C. The temperature ranges used in any of the embodiments herein may be any temperature range within the recited temperature ranges. Thus, in some embodiments, the bacteria may be cultured at temperatures such as about 4°C to about 140°C, about 4°C to about 80°C, about 4°C to about 40°C, about 16°C to about 40°C, about 16°C to about 60°C, about 22°C to about 37°C, about 22°C to about 45°C, or about 22°C to about 140°C.Similarly, a recited temperature range includes every individual temperature within said range. Thus, in some embodiments, the bacteria may be cultured at 4°C. In some embodiments, the bacteria may be cultured at 16°C. In some embodiments, the bacteria may be cultured at 22°C. In some embodiments, the bacteria may be cultured at 25°C. In some embodiments, the bacteria may be cultured at 30°C. In some embodiments, the bacteria may be cultured at 37°C. In some embodiments, bacteria may be cultured at 4°C, 5°C, 10°C, 15°C, 16°C, 17°C, 18°C, 19°C, 20°C, 21°C, 22°C, 23°C, 24°C, 25°C, 26°C, 27°C, 28°C, 29°C, 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, 38°C, 39°C, 40°C, 41°C, 42°C, 43°C, 44°C, 45°C, 50°C, 55°C, 60°C, 65°C, 70°C, 75°C, 80°C, 85°C, 90°C, 95°C, 100°C, 105°C, 110°C 115°C, 120°C, 125°C, 130°C, 135°C, 140°C, or any temperature in between the recited temperatures. Furthermore, those skilled in the art will recognize that further modifications to growth conditions may be necessary depending on the bacterial strain utilized and the therapeutic protein being produced. Such modifications are within the scope of the present application. Through various intracellular mechanisms, the payload protein is secreted extracellularly, with or without part or all of the synthetic preprotein signal peptide to which it is fused. In either case, secretion of the payload protein by the host bacterium results in accumulation of the payload protein in the surrounding medium, from which it may then be recovered, isolated, and / or quantified. Thus, in some embodiments, the method for producing a therapeutic protein further comprises recovering or purifying the payload protein from the medium. Techniques for isolating and purifying payload proteins from the medium are known in the art, and all such methods are within the scope of the present disclosure.

[0333] In some embodiments, the modified bacteria may be grown in any volume of medium. One of skill in the art will recognize that the volume of medium required for bacterial growth depends on the amount of payload protein desired to be produced. Thus, in some embodiments, the bacteria are cultured in volumes ranging from about 0.005 L to about 1,000,000 L or more. In some embodiments, the bacteria are cultured in a volume of at least 0.005 L. In some embodiments, the bacteria are cultured in volumes of about 0.005 L, 0.05 L, 0.5 L, 1 L, 2 L, 3 L, 4 L, 5 L, 10 L, 20 L, 30 L, 40 L, 50 L, 100 L, 1,000 L, 10,000 L, 100,000 L, or 1,000,000 L or more. In some embodiments, the bacteria may be cultured in any volume between any of the listed volumes or greater. Additionally, bacteria may be grown in a continuous culture system, whereby a portion of the bacterial culture is inoculated into fresh growth broth and the culture continued. It should be understood that the volumes listed should not be construed as limiting in any way, and that bacteria may be grown in any volume suitable for payload protein production.

[0334] In some embodiments, methods for producing a protein in a subject are provided. In some embodiments, the methods include administering a bacterium described herein to a subject. In some embodiments, the bacterium comprises a polypeptide provided herein. In some embodiments, the polypeptide comprises the formula X1-Z1, where X1 is a preprotein signal peptide provided herein and Z1 is a payload protein provided herein. In some embodiments, X1 comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:148, or SEQ ID NO:180, as provided herein. In some embodiments, Z1 is a payload protein provided herein. In some embodiments, Z1 is a payload protein selected from the group consisting of an antimicrobial peptide provided herein, catestatin, CRAMP, hBD1, hBD3, β-catethritin, dermaseptin, IL-10, IL-22, ClbS, TIGIT, TFF1, TFF2, TFF3, an anti-CCL2 antibody, an anti-CCL2 antibody fragment, an anti-IL23 antibody, an anti-IL23 antibody fragment, an anti-TNFα antibody, an anti-TNFα antibody fragment, GLP2, IGF1, leptin, and casein. In some embodiments, Z1 is a payload protein selected from the group consisting of an anti-TNFα antibody, an anti-TNFα antibody fragment, IL-10, IL-22, hBD3, leptin, and β-casein. In some embodiments, Z1 is an anti-TNFα antibody. In some embodiments, Z1 is an anti-TNFα antibody fragment. In some embodiments, Z1 is IL-10. In some embodiments, Z1 is IL-22. In some embodiments, Z1 is hBD3. In some embodiments, Z1 is leptin. In some embodiments, Z1 is β-casein. In some embodiments, the bacteria are administered orally. In some embodiments, the bacteria are administered topically. The bacteria of the present disclosure can survive in the environment in which they are administered.Thus, the bacteria continue to produce the payload protein when administered to a subject, thereby producing this protein in the subject.

[0335] In some embodiments, methods are provided for treating a disease or disorder in a subject in need thereof. In some embodiments, the method comprises administering a therapeutically effective amount of a bacterium described herein, or a pharmaceutical composition described herein comprising the bacterium, to the subject. The bacterium or a pharmaceutical composition comprising the bacterium may be administered via any suitable route known in the art. In some embodiments, the bacterium is administered orally or topically. In some embodiments, the bacterium i...

Claims

1. A preprotein signal peptide comprising an amino acid sequence having at least 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:148, or SEQ ID NO:

180.

2. X １ -Z １ A polypeptide comprising the formula: In the formula, X １ is the preprotein signal peptide, Z １ is a payload protein.

3. In the formula, X １ 3. The polypeptide of claim 2, comprising an amino acid sequence having at least 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:148, or SEQ ID NO:

180.

4. During the ceremony, Z １ The polypeptide of claim 2 or 3, wherein the polypeptide is selected from the group consisting of antimicrobial peptides, catestatin, CRAMP, hBD1, hBD3, β-catethritin, dermaseptin, IL-10, IL-22, ClbS, TIGIT, TFF1, TFF2, TFF3, anti-CCL2 antibody, anti-CCL2 antibody fragment, anti-IL23 antibody, anti-IL23 antibody fragment, anti-TNFα antibody, anti-TNFα antibody fragment, GLP2, IGF1, leptin, and casein.

5. In the formula, X １ comprises the amino acid sequence of SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:14, and Z １ The polypeptide of claim 2 , wherein said polypeptide comprises an anti-TNFα antibody or an anti-TNFα antibody fragment.

6. In the formula, X １ comprises the amino acid sequence of SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:14, and Z １ 6. The polypeptide of claim 5, comprising an amino acid sequence having at least 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, SEQ ID NO:120, or a fragment thereof.

7. In the formula, X １ The polypeptide of claim 5 or 6, comprising the amino acid sequence of SEQ ID NO:

6.

8. In the formula, X １ The polypeptide of claim 5 or 6, comprising the amino acid sequence of SEQ ID NO:

7.

9. In the formula, X １ The polypeptide of claim 5 or 6, comprising the amino acid sequence of SEQ ID NO:

14.

10. In the formula, X １ comprises the amino acid sequence of SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:148, or SEQ ID NO:180; Z １ The polypeptide of claim 2, wherein said polypeptide comprises leptin.

11. In the formula, X １ comprises the amino acid sequence of SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:148, or SEQ ID NO:180; Z １ 11. The polypeptide of claim 10, comprising an amino acid sequence having at least 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO:

25.

12. In the formula, X １ The polypeptide of claim 10 or 11, comprising the amino acid sequence of SEQ ID NO:

6.

13. In the formula, X １ The polypeptide of claim 10 or 11, comprising the amino acid sequence of SEQ ID NO:

7.

14. In the formula, X １ The polypeptide of claim 10 or 11, comprising the amino acid sequence of SEQ ID NO:

14.

15. In the formula, X １ The polypeptide of claim 10 or 11, comprising the amino acid sequence of SEQ ID NO:

17.

16. In the formula, X １ The polypeptide of claim 10 or 11, comprising the amino acid sequence of SEQ ID NO:

18.

17. In the formula, X １ The polypeptide of claim 10 or 11, comprising the amino acid sequence of SEQ ID NO:

19.

18. In the formula, X １ The polypeptide of claim 10 or 11, comprising the amino acid sequence of SEQ ID NO:

47.

19. In the formula, X １ The polypeptide of claim 10 or 11, comprising the amino acid sequence of SEQ ID NO:

48.

20. In the formula, X １ The polypeptide of claim 10 or 11, comprising the amino acid sequence of SEQ ID NO:

148.

21. In the formula, X １ The polypeptide of claim 10 or 11, comprising the amino acid sequence of SEQ ID NO:

180.

22. In the formula, X １ comprises the amino acid sequence of SEQ ID NO: 20, and Z １ The polypeptide of claim 2, wherein said polypeptide comprises IL-10.

23. In the formula, X １ comprises the amino acid sequence of SEQ ID NO: 20, and Z １ 23. The polypeptide of claim 22, comprising an amino acid sequence having at least 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO:24, SEQ ID NO:283, SEQ ID NO:284, or SEQ ID NO:

285.

24. In the formula, X １ comprises the amino acid sequence of SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:148, or SEQ ID NO:180; Z １ The polypeptide of claim 2, wherein said polypeptide comprises IL-22.

25. In the formula, X １ comprises the amino acid sequence of SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:148, or SEQ ID NO:180; Z １ 25. The polypeptide of claim 24, comprising an amino acid sequence having at least 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO:22, SEQ ID NO:59, or SEQ ID NO:

282.

26. In the formula, X １ 26. The polypeptide of claim 24 or 25, comprising the amino acid sequence of SEQ ID NO:

7.

27. In the formula, X １ 26. The polypeptide of claim 24 or 25, comprising the amino acid sequence of SEQ ID NO:

14.

28. In the formula, X １ 26. The polypeptide of claim 24 or 25, comprising the amino acid sequence of SEQ ID NO:

148.

29. In the formula, X １ 26. The polypeptide of claim 24 or 25, comprising the amino acid sequence of SEQ ID NO:

180.

30. X １ -Z １ A polypeptide comprising the formula: X １ is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:14; Z １ is a payload protein comprising an anti-TNFα antibody or an anti-TNFα antibody fragment.

31. During the ceremony, Z １ 31. The polypeptide of claim 30, wherein said polypeptide comprises an amino acid sequence selected from SEQ ID NO:21, SEQ ID NO:118, SEQ ID NO:119, or SEQ ID NO:

120.

32. X １ -Z １ A polypeptide comprising the formula: In the formula, X １ is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:148, or SEQ ID NO:180; Z １ is a payload protein comprising leptin.

33. During the ceremony, Z １ The polypeptide of claim 32, comprising the amino acid sequence of SEQ ID NO:

25.

34. X １ -Z １ A polypeptide comprising the formula: X １ is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO: 20; Z １ is a payload protein comprising IL-10.

35. During the ceremony, Z １ 35. The polypeptide of claim 34, comprising the amino acid sequence of SEQ ID NO:24, SEQ ID NO:283, SEQ ID NO:284, or SEQ ID NO:

285.

36. X １ -Z １ A polypeptide comprising the formula: X １ is a preprotein signal peptide comprising the amino acid sequence of SEQ ID NO:7, SEQ ID NO:14, SEQ ID NO:148, or SEQ ID NO:180; Z １ is a payload protein comprising IL-22.

37. During the ceremony, Z １ 37. The polypeptide of claim 36, wherein said polypeptide comprises the amino acid sequence of SEQ ID NO:22, SEQ ID NO:59, or SEQ ID NO:

282.

38. In the formula, X １ and Z １ The polypeptide according to any one of claims 2 to 37, wherein said polypeptides are linked by a linker polypeptide.

39. A nucleic acid molecule encoding the polypeptide of any one of claims 2 to 38.

40. X １ -Z １ 1. A bacterium comprising a heterologous nucleic acid molecule encoding a polypeptide comprising the formula: X １ is the preprotein signal peptide of claim 1, Z １ is a payload protein.

41. During the ceremony, Z １ The bacterium according to claim 40, wherein the antibody is selected from the group consisting of catestatin, CRAMP, hBD1, hBD3, β-catethritin, dermaseptin, IL-10, IL-22, ClbS, TIGIT, TFF1, TFF2, TFF3, anti-CCL2 antibody, anti-CCL2 antibody fragment, anti-IL23 antibody, anti-IL23 antibody fragment, anti-TNFα antibody, anti-TNFα antibody fragment, GLP2, IGF1, leptin, and casein.

42. Formula X １ -Z １ The bacterium according to claim 40, comprising a polypeptide according to any one of claims 2 to 38.

43. The bacterium according to any one of claims 40 to 42, wherein the bacterium is a Lactobacillus.

44. A pharmaceutical composition comprising the bacterium according to any one of claims 40 to 43 and a pharmaceutically acceptable carrier.

45. 1. A method for producing a payload protein, comprising: i) transfecting a bacterium with a nucleic acid molecule encoding a polypeptide according to any one of claims 2 to 38 to produce a bacterium comprising said nucleic acid molecule; ii) culturing the bacterium containing the nucleic acid molecule under conditions sufficient for the bacterium to grow, wherein the bacterium expresses and secretes the payload protein.

46. 46. ​​The method of claim 45, wherein the bacterium is Lactobacillus.

47. 45. A method for producing a payload protein in a subject, the method comprising administering to the subject the bacterium of any one of claims 40 to 43, or the pharmaceutical composition of claim 44, wherein the bacterium is live, and wherein the live bacterium produces the protein in the subject.

48. 45. A method for treating a disease or disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the bacterium of any one of claims 40 to 43 or the pharmaceutical composition of claim 44.

49. 49. The method of claim 48, wherein the disease or disorder is selected from inflammation, chronic inflammation, colorectal cancer, periodontitis, oral mucositis, infection, pathogenic E. coli, inflammatory bowel disease, ulcerative colitis, Crohn's disease, type 2 diabetes, obesity, lipodystrophy, hypertriglyceridemia, systemic lupus erythematosus, fatty liver, bone mineral density loss, hypothalamic hypogonadism, infertility, hypothalamic amenorrhea, autoimmune disease, cancer, NAFLD, NASH, infection, sepsis, or growth hormone disorder.

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