Carotenoids for treating or preventing nausea

Abstract

To provide a technique relating to treatment of nausea and / or vomiting.SOLUTION: Administering to a subject in need thereof a therapeutically effective amount of a C50 carotenoids compound. The C50 carotenoids compound is selected from the group consisting of decaprenoxanthin, C50 - astaxanthin, C50 - beta-carotene, C50 - carotene (n = 3) (1616 - diisopentenylphytoene), C50 - zeaxanthin, C50 - caroxanthin, C50 - nostoxanthin, sarsinaxanthin, sarprenoxanthin, acyclic C50 carotenoids bacterioruberin, C50 - canthaxanthin, - lycopene, - phytoene, and combinations thereof. C50 C50.SELECTED DRAWING: Figure 1A

Description

[Technical Field]

[0001] Priority claims This application claims the benefit of U.S. Provisional Application No. 62,802,398, filed February 7, 2019, the entire contents of which are incorporated herein by reference. O

[0002] Federally funded research or development This invention was made with government support under Grant No. AG043184 awarded by the National Institutes of Health. The government has certain rights in this invention. [Background technology]

[0003] background Chemotherapy-induced nausea and vomiting (CINV) affects 70-80% of patients receiving chemotherapy and is the leading cause of discontinuation of cancer treatment. Summary of the Invention

[0004] overview The present disclosure provides techniques for the treatment and / or prevention of certain diseases, disorders, and / or conditions, and further provides techniques for evaluating one or more properties of agents useful for such treatment and / or prevention.

[0005] In some embodiments, the provided technology relates to the treatment of nausea and / or vomiting, and in some embodiments, particularly induced (e.g., chemotherapy-induced) nausea and / or vomiting. Alternatively, or in addition, in some embodiments, the provided technology relates to the treatment or prevention of one or more eating disorders (e.g., anorexia nervosa).

[0006] Nausea and / or vomiting are experienced in some diseases and disorders, and can also result from therapeutic treatment such as chemotherapy.No matter what situation causes nausea and / or vomiting, both are generally considered unpleasant and undesirable.Therefore, subjects try to avoid nausea and / or vomiting; this avoidance can lead to, for example, non-compliance with therapeutic treatment, for example, when patients find the relationship between such treatment and the nausea and / or vomiting they experience.

[0007] The present disclosure recognizes that certain biological pathways involved in monitoring and / or responding to toxins (e.g., toxin detoxification pathways), particularly certain signaling pathways that convert the detection of translational defects into the induction of detoxification genes, are conserved across animal lineages and can also suppress aberrant human xenobiotic responses; the present disclosure teaches that such agents may have therapeutic potential, particularly in the treatment of certain diseases, disorders, or conditions including, for example, nausea and / or vomiting (e.g., chemotherapy-induced nausea, which is a major problem in cancer therapy), and / or eating disorders such as anorexia nervosa.

[0008] Thus, the present disclosure provides model systems (e.g., C. elegans) for characterizing agents for their utility as therapeutic agents described herein. Further, the present disclosure describes the use of such systems to identify and / or characterize certain useful such agents.

[0009] Among other things, the present disclosure teaches that certain carotenoid compounds (e.g., certain C50 carotenoid compounds) are useful for treating and / or preventing diseases, disorders and / or conditions such as nausea and / or vomiting (e.g., induced nausea, such as chemotherapy-induced nausea), and / or one or more eating disorders (e.g., anorexia nervosa).

[0010] For example, the present disclosure provides that certain carotenoid compounds (e.g., certain C50 carotenoid compounds) inhibit detoxification pathways, including those activated in response to translation defects induced by toxins and / or by mutations in translation components. The present disclosure recognizes that carotenoids are generally well tolerated by mammals, including humans, and teaches that carotenoids are an attractive class of compounds for the therapeutic uses described herein (e.g., for the treatment and / or prevention of nausea and / or vomiting and / or one or more eating disorders).

[0011] The present disclosure specifically recognizes that certain C50 carotenoid compounds are naturally produced by microorganisms (e.g., as reviewed by Hencke et al., "C50 Carotenoids: Occurrence, Biosynthesis, Glycosylation, and Metabolic Engineering for Their Overproduction," Chapter 5 of Bio-pigmentation and Biotechnological Implementations, Ed. Singh, Wiley & Sons, 2017). In some embodiments, delivery of carotenoid compounds (e.g., C50 carotenoid compounds) for therapeutic uses described herein can be achieved by administration of a composition that is or includes a microorganism that produces one or more carotenoid compounds of interest, or an extracted or purified component thereof. In some embodiments, such administration can be viable (e.g., microorganism), and in certain embodiments, can achieve establishment of the administered microorganism in the recipient (e.g., as part of the recipient's microbiome).

[0012] Among other things, the present disclosure recognizes that embodiments involving administration of microorganisms, particularly viable (e.g., living) microorganisms, may offer certain advantages such as, for example, reduced dosing (e.g., reduced dosing frequency, duration of dosing, total number of doses administered, amount and / or concentration of administered doses, and / or combinations thereof), reduced cost, long-term effectiveness, etc.

[0013] However, those skilled in the art will recognize, upon reading this disclosure, that delivery of carotenoid compounds (e.g., C50 carotenoid compounds) for the treatments described herein is not limited to delivery of microorganisms, or even extracts and / or components thereof; rather, useful carotenoid compounds (e.g., as described herein) can be prepared in whole or in part by chemical synthesis and / or purified from microbial sources (e.g., cultured microbial cells, which can be naturally occurring and / or genetically or otherwise engineered cells).

[0014] Those skilled in the art will further recognize that any of a variety of delivery routes and / or forms can be utilized to administer compositions that deliver the useful carotenoid compounds described herein (e.g., that are or include microorganisms and / or extracts or components thereof and / or one or more pure carotenoid compounds described herein). In many embodiments, the compositions are administered orally (e.g., via pills, tablets, capsules, powders, lozenges, troches, syrups, elixirs, etc.). In some embodiments, oral administration is via a nutritional source such as food or drink.

[0015] One challenge associated with developing useful treatments for nausea and / or vomiting has been the lack of animal models. The present disclosure describes that Caenorhabditis elegans can provide an effective model for nausea and / or vomiting. Among other things, the present disclosure provides that C. elegans can be useful for characterizing (e.g., screening) agents to evaluate their effects on and / or utility in treating nausea and / or vomiting. For example, microbial toxins and virulence factors often target the translation machinery. C. elegans responds to translation defects (e.g., as may result from exposure to toxins and / or mutations in translation components) by inducing detoxification and defense response genes. According to the present disclosure, agents that inhibit this induction may be useful for treating nausea and / or vomiting, and evaluation of such inhibition can be useful for characterizing (e.g., screening) such agents.

[0016] Furthermore, the present disclosure demonstrates that certain carotenoid compounds (e.g., certain C50 carotenoid compounds) can inhibit the C. elegans translation defect monitoring and response pathway (e.g., the C. elegans xenobiotic detoxification response to translation defects); such carotenoid compounds may be useful in accordance with the present disclosure in therapeutic applications, such as for treating nausea and / or vomiting. For example, the present disclosure specifically describes that the C50 carotenoid compound produced by Kocuria rhizophila inhibits the C. elegans translation defect monitoring and response pathway. Among other things, the present disclosure describes genetic analyses identifying the biosynthetic pathway for this carotenoid as mediating the suppression of the C. elegans translation toxin defense response. Furthermore, the present disclosure describes that K. rhizophila extracts (i) mimic the suppression of the xenobiotic detoxification response of C. elegans to translation defects; and (ii) restore the ability of carotenoid mutants of K. rhizophila to inhibit such detoxification responses to translation defects.

[0017] Additionally, the present disclosure describes that other carotenoid compounds (e.g., C50 carotenoid compounds produced by other bacterial species) also inhibit C. elegans translation defect monitoring and response pathways (e.g., the C. elegans xenobiotic detoxification response to translation defects). For example, the present disclosure describes that C. glutamicum, which produces the C50 carotenoid decaprenoxanthin, also inhibits the C. elegans detoxification response, and further describes that carotenoid biosynthesis mutants of C. glutamicum are defective in this inhibition.

[0018] Furthermore, the present disclosure describes that yet another bacterial species, Arthrobacter arilaitensis, which also produces C50 carotenoids (specifically, decaprenoxanthin), also inhibits the detoxification response of C. elegans.

[0019] Without wishing to be bound by any particular theory, the present disclosure proposes that the carotenoid compounds described herein (e.g., C50 carotenoid compounds) suppress the induction of xenobiotic detoxification by inhibiting the bile acid signaling pathway in C. elegans, which converts the detection of translation defects into the induction of detoxification genes. Suppression of translation surveillance by the carotenoid compounds described herein (e.g., C50 carotenoid compounds) abolishes the drug detoxification response, resulting in enhanced efficacy of translation inhibitors. Thus, in some embodiments, carotenoid compounds useful according to the present disclosure may be characterized by their ability to increase the efficacy of translation inhibitors. For example, in some embodiments, useful carotenoid compounds are characterized by an enhancement of one or more characteristics of the effect of a translation inhibitor on C. elegans when such a carotenoid compound is contacted with C. elegans in the presence of a translation inhibitor, compared to that observed under otherwise equivalent conditions (e.g., the presence of the same translation inhibitor at the same concentration) in the absence of the carotenoid compound.

[0020] The present disclosure also demonstrates that certain carotenoid compounds (e.g., certain C50 carotenoid compounds) inhibit the coupling of translational surveillance to food aversion behavior in C. elegans, which is normally induced by translation inhibitors. Thus, in some embodiments, carotenoid compounds useful according to the present disclosure may be characterized by their effect on food aversion behavior in C. elegans in the presence of a toxin. For example, in some embodiments, useful carotenoid compounds may be characterized by altering one or more characteristics of the effect of a toxin that targets protein translation on food avoidance behavior in C. elegans when such compounds are contacted with C. elegans compared to that observed under otherwise equivalent conditions (e.g., the presence of the same toxin at the same concentration) in the absence of the carotenoid compound. In some embodiments, agents (e.g., carotenoid compounds such as C50 carotenoid compounds) shown to affect food aversion behavior described herein may be particularly useful for treating one or more food aversion disorders, such as anorexia nervosa.

[0021] Therefore, provided herein is a method for treating nausea and / or vomiting or reducing food aversion in a subject.The method comprises administering a therapeutically effective amount of a C50 carotenoid compound to a subject in need thereof.In some embodiments, the subject has or is at risk of developing nausea and / or vomiting associated with chemotherapy or radiation, such as chemotherapy-induced nausea and vomiting (CINV) or radiation-induced nausea and vomiting (RINV).

[0022] In some embodiments, the subject has or is at risk of developing post-operative nausea and vomiting (PONV).

[0023] In some embodiments, the C50 carotenoid compound is selected from the group consisting of decaprenoxanthin, C50-astaxanthin, C50-β-carotene, C50-carotene (n=3) (16,16-diisopentenylphytoene), C50-zeaxanthin, C50-caroxanthin, C50-nostoxanthin, sarcinaxanthin, sarprenoxanthin, the acyclic C50 carotenoid bacterioruberin, C50-canthaxanthin, C50-lycopene, C50-phytoene, and combinations thereof. In some embodiments, the C50 carotenoid compound is decaprenoxanthin.

[0024] In some embodiments, the administering step comprises administering a composition that is or comprises: (i) a microorganism or a component thereof that synthesizes C50 carotenoid compounds, (ii) an extract from a microorganism that synthesizes C50 carotenoid compounds, (iii) an extracted carotenoid compound, or (iv) a combination thereof. In some embodiments, the microorganism that synthesizes C50 carotenoid compounds is viable or alive. In some embodiments, the administering step comprises administering a sufficient amount of the microorganism to colonize the microbiome of the subject. In some embodiments, the composition comprises a culture of the microorganism or is prepared from a culture of the microorganism. In some embodiments, the microorganism is a strain found in nature. In some embodiments, the microorganism is an engineered microorganism. In some embodiments, the engineered microorganism comprises a genetic mutation compared to an otherwise identical reference microorganism such that the engineered microorganism produces C50 carotenoid compounds at an absolute or relative level that differs from that of the reference microorganism.

[0025] In some embodiments, the administering step comprises administering a composition containing or delivering a synthesized C50 carotenoid compound. In some embodiments, the microorganism that synthesizes the C50 carotenoid compound is selected from the group consisting of Kocuria rhizophila, Corynebacterium glutamicum, Arthrobacter alilactensis, and combinations thereof.

[0026] Also provided herein is a therapeutic composition for oral delivery comprising a therapeutically effective amount of a C50 carotenoid compound and a pharmaceutically acceptable carrier.

[0027] In some embodiments, the composition comprises a microorganism that synthesizes a C50 carotenoid compound. In some embodiments, the microorganism is a cultured microorganism. In some embodiments, the microorganism is an engineered microorganism. In some embodiments, the engineered microorganism comprises a genetic mutation relative to an otherwise identical reference microorganism such that the engineered microorganism produces a C50 carotenoid compound at an absolute or relative level that differs from that of the reference microorganism. In some embodiments, the microorganism is live or viable. In some embodiments, the microorganism is killed. In some embodiments, the microorganism is selected from the group consisting of Kocuria rhizophila, Corynebacterium glutamicum, Arthrobacter alilactensis, and combinations thereof.

[0028] In some embodiments, the C50 carotenoid compound is at least 20% w / w of the composition.

[0029] In some embodiments, the C50 carotenoid compound is purified.

[0030] In some embodiments, the C50 carotenoid compound has a chemical structure found in nature.

[0031] In some embodiments, the C50 carotenoid compound is an analog of a reference C50 carotenoid compound found in nature.

[0032] In some embodiments, the C50 carotenoid compound is selected from the group consisting of decaprenoxanthin, C50-astaxanthin, C50-β-carotene, C50-carotene (n=3) (16,16-diisopentenylphytoene), C50-zeaxanthin, C50-caroxanthin, C50-nostoxanthin, sarcinaxanthin, sarprenoxanthin, the acyclic C50 carotenoid bacterioruberin, C50-canthaxanthin, C50-lycopene, C50-phytoene, and combinations thereof. In some embodiments, the C50 carotenoid compound is decaprenoxanthin.

[0033] In some embodiments, the therapeutic composition is a liquid, syrup, tablet, lozenge, gummy, capsule, powder, gel, or film.

[0034] Furthermore, the present specification provides a method for preparing a therapeutic composition.For example, the method can include combining a pharmaceutically acceptable carrier with a C50 carotenoid compound; and formulating the combination into a therapeutic composition.

[0035] In some embodiments, the combining step comprises combining a pharmaceutically acceptable carrier with a microorganism that synthesizes a C50 carotenoid compound.

[0036] In some embodiments, the combining step comprises combining a pharmaceutically acceptable carrier with a chemically synthesized C50 carotenoid compound.

[0037] Also provided herein is a method for treating nausea and / or vomiting or reducing food aversion in a subject, the method comprising administering to a subject in need thereof (i) a microorganism that synthesizes a C50 carotenoid compound or a component thereof, (ii) an extract of a microorganism that synthesizes a C50 carotenoid compound, or (iii) an extracted C50-carotenoid compound, or (iv) a combination thereof.

[0038] In some embodiments, the subject has or is at risk of developing nausea and / or vomiting associated with chemotherapy or radiation.

[0039] In some embodiments, the subject has or is at risk of developing chemotherapy-induced nausea and vomiting (CINV) or radiation-induced nausea and vomiting (RINV). In some embodiments, the subject has or is at risk of developing post-operative nausea and vomiting (PONV).

[0040] In some embodiments, the microorganism that synthesizes a C50 carotenoid compound is selected from the group consisting of Kocuria rhizophila, Corynebacterium glutamicum, Arthrobacter alilactensis, and combinations thereof.

[0041] Further provided herein are uses of C50 carotenoid compounds for treating nausea and / or vomiting or reducing food aversions in a subject in need thereof, and C50 carotenoid compounds for use in treating nausea and / or vomiting or reducing food aversions in a subject in need thereof.

[0042] Also provided are microorganisms that synthesize C50 carotenoid compounds for treating nausea and / or vomiting or reducing food aversions in a subject in need thereof, and uses of microorganisms that synthesize C50 carotenoid compounds for treating nausea and / or vomiting or reducing food aversions.

[0043] In some embodiments, the microorganism that synthesizes a C50 carotenoid compound is selected from the group consisting of Kocuria rhizophila, Corynebacterium glutamicum, Arthrobacter alilactensis, and combinations thereof.

[0044] Also provided herein are methods for evaluating carotenoid compounds for anti-nausea and / or anti-emetic activity, the methods including (i) contacting a system with a carotenoid compound; and (ii) determining whether the carotenoid compound alters a characteristic of the system, the characteristic being associated with nausea and / or vomiting.

[0045] In some embodiments, the determining step comprises comparing the characteristics before and after performing the contacting step.

[0046] In some embodiments, the determining step comprises comparing the characteristic to an equivalent reference after the contacting step.

[0047] In some embodiments, the equivalent reference is a historical reference.

[0048] In some embodiments, the comparable reference is a negative control reference.

[0049] In some embodiments, the comparable reference is a positive control reference.

[0050] In some embodiments, the system is or comprises C. elegans.

[0051] In some embodiments, the characteristic is level of food aversion.

[0052] In some embodiments, the feature is the level or activity of a nucleic acid or protein, or a form thereof.

[0053] In some embodiments, the trait is or comprises an aspect of a xenobiotic detoxification response.

[0054] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this invention belongs.Methods and materials for use in the present invention are described herein; other suitable methods and materials known in the art can also be used.Materials, methods and examples are only illustrative and are not intended to be limiting.All publications, patent applications, patents, sequences, database entries and other references mentioned herein are incorporated by reference in their entirety.In case of discrepancy, the present specification, including definitions, shall prevail.

[0055] [The present invention 1001] administering a therapeutically effective amount of a C50 carotenoid compound to a subject in need thereof. 10. A method for treating nausea and / or vomiting in a subject, comprising: [The present invention 1002] 1002. The method of claim 1001, wherein said subject has or is at risk of developing nausea and / or vomiting associated with chemotherapy or radiation. [The present invention 1003] The method of any one of claims 1001 to 1002, wherein said subject has or is at risk of developing chemotherapy-induced nausea and vomiting (CINV) or radiation-induced nausea and vomiting (RINV). [The present invention 1004] 1001. The method of claim 1001, wherein said subject has or is at risk of developing post-operative nausea and vomiting (PONV). [The present invention 1005] Any of the methods of inventions 1001 to 1004, wherein the C50 carotenoid compound is selected from the group consisting of decaprenoxanthin, C50-astaxanthin, C50-β-carotene, C50-carotene (n=3) (16,16-diisopentenylphytoene), C50-zeaxanthin, C50-caloxanthin, C50-nostoxanthin, sarcinaxanthin, sarprenoxanthin, the acyclic C50 carotenoid bacterioruberin, C50-canthaxanthin, C50-lycopene, C50-phytoene, and combinations thereof. [The present invention 1006] 1005. The method of claim 10, wherein the C50 carotenoid compound is decaprenoxanthin. [The present invention 1007] Any of the methods of inventions 1001 to 1006, wherein the administration step comprises administering (i) a microorganism that synthesizes a C50 carotenoid compound or a component thereof, (ii) an extract of a microorganism that synthesizes a C50 carotenoid compound, (iii) an extracted C50-carotenoid compound, or (iv) a composition that is a combination thereof; or a composition containing them. [The present invention 1008] 1007. The method of claim 1007, wherein said microorganism that synthesizes C50 carotenoid compounds is viable or alive. [The present invention 1009] 1008. The method of claim 10, wherein said administering step comprises administering said microorganism in an amount sufficient to colonize the microbiome of said subject. [The present invention 1010] 1009. The method of any of claims 1007 to 1009, wherein said composition comprises or is prepared from a culture of said microorganism. [The present invention 1011] The method of claim 10, wherein said microorganism is a strain found in nature. [The present invention 1012] The method of any one of claims 1007 to 1009 or 1010, wherein said microorganism is an engineered microorganism. [The present invention 1013] The method of claim 10, wherein the engineered microorganism comprises a genetic mutation relative to an otherwise identical reference microorganism such that the engineered microorganism produces the C50 carotenoid compound at an absolute or relative level that differs from that of the reference microorganism. [The present invention 1014] The method of any of claims 1001 to 1013, wherein said administering step comprises administering a composition that contains or delivers a synthetic C50 carotenoid compound. [The present invention 1015] 1007. The method of claim 1007, wherein the microorganism that synthesizes the C50 carotenoid compound is selected from the group consisting of Kocuria rhizophila, Corynebacterium glutamicum, Arthrobacter arilaitensis, and combinations thereof. [The present invention 1016] administering a therapeutically effective amount of a C50 carotenoid compound to a subject in need thereof. 10. A method for reducing food aversions in a subject, comprising: [The present invention 1017] The method of claim 1016, wherein said subject has or is at risk of developing nausea and vomiting associated with chemotherapy or radiation. [The present invention 1018] The method of any one of claims 1016 to 1017, wherein said subject has or is at risk of developing chemotherapy-induced nausea and vomiting (CINV) or radiation-induced nausea and vomiting (RINV). [The present invention 1019] The method of claim 1016, wherein said subject has or is at risk of developing post-operative nausea and vomiting (PONV). [The present invention 1020] Any of the methods of claims 1016 to 1019, wherein the C50 carotenoid compound is selected from the group consisting of decaprenoxanthin, C50-astaxanthin, C50-β-carotene, C50-carotene (n=3) (16,16-diisopentenylphytoene), C50-zeaxanthin, C50-caloxanthin, C50-nostoxanthin, sarcinaxanthin, sarprenoxanthin, the acyclic C50 carotenoid bacterioruberin, C50-canthaxanthin, C50-lycopene, C50-phytoene, and combinations thereof. [The present invention 1021] The method of claim 1020, wherein the C50 carotenoid compound is decaprenoxanthin. [The present invention 1022] Any of the methods of inventions 1016 to 1021, wherein the administration step comprises administering (i) a microorganism that synthesizes a C50 carotenoid compound or a component thereof, (ii) an extract of a microorganism that synthesizes a C50 carotenoid compound, (iii) an extracted C50-carotenoid compound, or (iv) a composition that is a combination thereof; or a composition containing them. [The present invention 1023] The method of claim 1022, wherein the microorganism that synthesizes the C50 carotenoid compound is viable or alive. [The present invention 1024] The method of claim 1023, wherein said administering step comprises administering said microorganism in an amount sufficient to colonize the microbiome of said subject. [The present invention 1025] The method of any of claims 1022 to 1024, wherein said composition comprises or is prepared from a culture of said microorganism. [The present invention 1026] 1025. The method of claim 1025, wherein the microorganism is a strain found in nature. [The present invention 1027] 1025. The method of any one of claims 1022 to 1024, wherein the microorganism is an engineered microorganism. [The present invention 1028] The method of claim 1027, wherein said engineered microorganism comprises a genetic mutation relative to an otherwise identical reference microorganism such that said engineered microorganism produces said C50 carotenoid compound at an absolute or relative level that differs from that of the reference microorganism. [The present invention 1029] 1029. The method of any of claims 1016 to 1028, wherein said administering step comprises administering a composition that contains or delivers a synthetic C50 carotenoid compound. [The present invention 1030] The method of claim 1022, wherein the microorganism that synthesizes the C50 carotenoid compound is selected from the group consisting of Kocuria rhizophila, Corynebacterium glutamicum, Arthrobacter alilactensis, and combinations thereof. [The present invention 1031] A therapeutic composition for oral delivery comprising a therapeutically effective amount of a C50 carotenoid compound and a pharmaceutically acceptable carrier. [The present invention 1032] 1031. A therapeutic composition of the present invention comprising a microorganism that synthesizes said C50 carotenoid compound. [The present invention 1033] The therapeutic composition of claim 1032, wherein the microorganism is a cultured microorganism. [The present invention 1034] The therapeutic composition of claim 1033, wherein said microorganism is an engineered microorganism. [This invention 1035] 1034. The therapeutic composition of the present invention, wherein said engineered microorganism comprises a genetic mutation relative to an otherwise identical reference microorganism such that said engineered microorganism produces said C50 carotenoid compound at an absolute or relative level that differs from that of the reference microorganism. [The present invention 1036] The therapeutic composition of any one of claims 1031 to 1034, which is a liquid, syrup, tablet, lozenge, gummy, capsule, powder, gel, or film. [This invention 1037] 1037. The therapeutic composition of any one of claims 1031 to 1036, wherein said C50 carotenoid compound is at least 20% w / w of said composition. [The present invention 1038] The therapeutic composition of any one of claims 1031 to 1037, wherein the C50 carotenoid compound is purified. [This invention 1039] 1039. The therapeutic composition of any one of claims 1031 to 1038, wherein the C50 carotenoid compound has a chemical structure found in nature. [The present invention 1040] 1039. The therapeutic composition of any of claims 1031 to 1039, wherein said C50 carotenoid compound is an analog of a reference C50 carotenoid compound found in nature. [The present invention 1041] The therapeutic composition of any of claims 1031 to 1040, wherein the C50 carotenoid compound is selected from the group consisting of decaprenoxanthin, C50-astaxanthin, C50-β-carotene, C50-carotene (n=3) (16,16-diisopentenylphytoene), C50-zeaxanthin, C50-caroxanthin, C50-nostoxanthin, sarcinaxanthin, sarprenoxanthin, the acyclic C50 carotenoid bacterioruberin, C50-canthaxanthin, C50-lycopene, C50-phytoene, and combinations thereof. [The present invention 1042] The therapeutic composition of the present invention 1041, wherein the C50 carotenoid compound is decaprenoxanthin. [This invention 1043] The therapeutic composition of any one of claims 1031 to 1042, wherein the microorganism is alive or viable. [This invention 1044] The therapeutic composition of any one of claims 1031 to 1043, wherein the microorganism has been killed. [This invention 1045] The therapeutic composition of any of claims 1031 to 1044, wherein the microorganism is selected from the group consisting of Kocuria rhizophila, Corynebacterium glutamicum, Arthrobacter alilactensis, and combinations thereof. [The present invention 1046] A method for producing a therapeutic composition according to any one of claims 1031 to 1045, comprising the following steps: combining a pharmaceutically acceptable carrier with a C50 carotenoid compound; and formulating the combination into a therapeutic composition. [This invention 1047] The method of claim 1046, wherein said combining step comprises combining a pharmaceutically acceptable carrier with a microorganism that synthesizes a C50 carotenoid compound. [This invention 1048] The method of claim 1046, wherein said combining step comprises combining a pharmaceutically acceptable carrier with a chemically synthesized C50 carotenoid compound. [This invention 1049] A method for treating nausea and / or vomiting in a subject, comprising: Administering (i) a microorganism that synthesizes C50 carotenoid compounds or a component thereof, (ii) an extract of a microorganism that synthesizes C50 carotenoid compounds, (iii) the extracted C50-carotenoid compounds, or (iv) a combination thereof to a subject in need thereof. [The present invention 1050] The method of claim 1049, wherein said subject has or is at risk of developing nausea and / or vomiting associated with chemotherapy or radiation. [This invention 1051] The method of any one of claims 1049 to 1050, wherein said subject has or is at risk of developing chemotherapy-induced nausea and vomiting (CINV) or radiation-induced nausea and vomiting (RINV). [This invention 1052] The method of claim 1049, wherein said subject has or is at risk of developing post-operative nausea and vomiting (PONV). [This invention 1053] Any of the methods of claims 1049 to 1052, wherein the microorganism that synthesizes the C50 carotenoid compound is selected from the group consisting of Kocuria rhizophila, Corynebacterium glutamicum, Arthrobacter alilactensis, and combinations thereof. [This invention 1054] A method for reducing food aversions in a subject, comprising: Administering (i) a microorganism that synthesizes C50 carotenoid compounds or a component thereof, (ii) an extract of a microorganism that synthesizes C50 carotenoid compounds, (iii) the extracted C50-carotenoid compounds, or (iv) a combination thereof to a subject in need thereof. [This invention 1055] The method of claim 1054, wherein said subject has or is at risk of developing nausea and vomiting associated with chemotherapy or radiation. [The present invention 1056] The method of any one of claims 1054 to 1055, wherein said subject has or is at risk of developing chemotherapy-induced nausea and vomiting (CINV) or radiation-induced nausea and vomiting (RINV). [This invention 1057] The method of claim 1054, wherein said subject has or is at risk of developing post-operative nausea and vomiting (PONV). [This invention 1058] Any of the methods of claims 1054 to 1057, wherein the microorganism that synthesizes the C50 carotenoid compound is selected from the group consisting of Kocuria rhizophila, Corynebacterium glutamicum, Arthrobacter alilactensis, and combinations thereof. [This invention 1059] Use of a C50 carotenoid compound for treating nausea and / or vomiting in a subject in need thereof. [The present invention 1060] Use of a microorganism that synthesizes a C50 carotenoid compound to treat nausea and / or vomiting in a subject in need thereof. [The present invention 1061] The use of the present invention 1060, wherein the microorganism that synthesizes the C50 carotenoid compound is selected from the group consisting of Kocuria rhizophila, Corynebacterium glutamicum, Arthrobacter alilactensis, and combinations thereof. [The present invention 1062] Use of a C50 carotenoid compound to reduce food aversion in a subject. [This invention 1063] Use of a microorganism that synthesizes C50 carotenoid compounds to reduce food aversions in a subject. [This invention 1064] The use of the present invention 1063, wherein the microorganism that synthesizes the C50 carotenoid compound is selected from the group consisting of Kocuria rhizophila, Corynebacterium glutamicum, Arthrobacter alilactensis, and combinations thereof. [This invention 1065] A method for evaluating carotenoid compounds for anti-nausea and / or anti-vomiting activity, comprising the steps of: (i) contacting the system with a carotenoid compound; (ii) determining whether the carotenoid compound alters a characteristic of the system, the characteristic being associated with nausea and / or vomiting. [The present invention 1066] 1065. The method of claim 1065, wherein said determining step comprises comparing said characteristics before and after performing said contacting step. [This invention 1067] 1065. The method of claim 1065, wherein said determining step comprises, after said contacting step, comparing said characteristic with an equivalent reference. [The present invention 1068] 1067. The method of claim 1067, wherein said equivalent reference is a historical reference. [The present invention 1069] 1067. The method of claim 1067, wherein said equivalent reference is a negative control reference. [The present invention 1070] 1067. The method of claim 1067, wherein said equivalent reference is a positive control reference. [This invention 1071] The method of any of claims 1065 to 1070, wherein the system is or comprises C. elegans. [This invention 1072] 1072. The method of any one of claims 1065 to 1071, wherein the characteristic is the level of food aversion. [This invention 1073] 1072. The method of any one of claims 1065 to 1071, wherein said characteristic is the level or activity of a nucleic acid or protein, or the form thereof. [This invention 1074] 1074. The method of claim 1073, wherein said characteristic is or comprises an aspect of a xenobiotic detoxification response. Other features and advantages of aspects of the present invention will become apparent from the following detailed description, drawings, and claims. [Brief explanation of the drawings]

[0056] [Figure 1A] Induction of pgp-5p::gfp was significantly reduced in eft-3(q145);pgp-5p::gfp animals fed K. rhizophila wild-type, whereas K. rhizophila crtEb(e17), K. rhizophila crtI(e10), and K. rhizophila crtYe(e2) mutants did not suppress GFP induction. [Figure 1B] Feeding K. rhizophila significantly reduced the induction of pgp-5p::gfp in eft-3(q145);pgp-5p::gfp animals, whereas expression of pgp-5p::gfp was unaffected in K. rhizophila mutants. Unpaired t-test, **P<0.01. Mean ± sd is shown. The number of animals analyzed per condition is indicated above each bar. ns is not significant compared to eft-3(q145);pgp-5p::gfp fed E. coli OP50. [Figure 1C] Discoloration phenotype of K. rhizophila mutants. [Figure 1D-1] Diagrammatic representation of mutations in the carotenoid cluster of K. rhizophylla . [Figure 1D-2] This is a continuation of Figure 1D-1. [Figure 1E] Putative C50 carotenoid biosynthetic pathway in K. rhizophila. [Figure 2A]A) Induction of pgp-5p::gfp was significantly reduced in eft-3(q145);pgp-5p::gfp animals fed C. glutamicum wild type, whereas C. glutamicum ΔcrtEb, ΔcrtI, ΔcrtY, and ΔcrtB mutants did not suppress GFP induction. [Figure 2B] Quantification of pgp-5p::gfp expression in eft-3(q145);pgp-5p::gfp animals fed C. glutamicum wild-type, ΔcrtEb, ΔcrtI, ΔcrtY, and ΔcrtB mutants. Unpaired t-test, ****P<0.0001. Mean ± sd shown. Number of animals analyzed per condition is indicated above each bar. ns not significant compared to eft-3(q145);pgp-5p::gfp fed E. coli OP50. [Figure 2C] Induction of pgp-5p::gfp was significantly reduced in eft-3(q145);pgp-5p::gfp animals fed A. arilaitensis wild-type. [Figure 2D] TLC of K. rhizophylla extract showing orange pigment. [Figure 2E] HPLC of K. rhizophila extract showing the absorbance of the orange pigment. The inset numbers indicate the elution times and absorbance of the various peaks from the extract. [Figure 2F] 750 μg / ml of K. rhizophila extract inhibited pgp-5p::gfp in eft-3(q145);pgp-5p::gfp animals. [Figure 2G] Quantification of pgp-5p::gfp expression in eft-3(q145);pgp-5p::gfp animals fed K. rhizophila wild-type, K. rhizophila crtEb(e17), K. rhizophila crtI(e10), and K. rhizophila crtEb(e6) with either control extract or K. rhizophila extract. [Figure 3A]Induction of pgp-5p::gfp in response to 10 mg / ml hygromycin was significantly reduced in animals fed K. rhizophila wild type, but GFP induction was not suppressed in K. rhizophila crtEb(e17) or K. rhizophila crtI(e10) mutants. [Figure 3B] Animals treated with K. rhizophila carotenoid extract were hypersensitive to hygromycin. Paired t-test, ****P<0.0001 compared to wild-type worms fed E. coli OP50 with solvent extract and hygromycin. Mean ± SD shown. Data collected from three independent trials with at least 20 animals per condition. ns not significant compared to wild-type worms fed E. coli OP50 with solvent extract without hygromycin. [Figure 3C] Animals treated with the carotenoid extract of K. rhizophila were hypersensitive to emetine. [Figure 3D] Animals treated with carotenoid extract of K. rhizophila were hypersensitive to cisplatin. [Figure 3E] Animals treated with the carotenoid extract of K. rhizophila did not avoid hygromycin compared to vehicle control and hygromycin treated animals. [Figure 3F] Animals treated with the carotenoid extract of K. rhizophylla did not avoid cisplatin compared to vehicle- and cisplatin-treated animals. Unpaired t-test, ****P<0.01, **P<0.0001. Mean ± sd shown. ns not significant. [Figure 4A] pgp-5p::gfp was constitutively induced in animals fed E. coli OP50 or K. rhizophila expressing ZIP-2::mCherry in the intestine under the control of the vha-6 promoter. [Figure 4B] Bile acid supplementation suppressed the K. rhizophila -induced pgp-5p::gfp activation defect in eft-3(q145);pgp-5p::gfp animals. [Figure 4C]Quantification of bile acid suppression of the K. rhizophila-induced pgp-5p::gfp activation defect in eft-3(q145);pgp-5p::gfp animals shown in Figure 4B. Unpaired t-test, ***P<0.0001. Mean ± sd is shown. The number of animals analyzed per condition is indicated above each bar. ns was not significant compared to eft-3(q145);pgp-5p::gfp fed E. coli OP50. [Figure 4D] lbp-5 RNAi, chc-1RNAi, fcho-1 RNAi, and dyn-1 RNAi suppressed the K. rhizophila -induced pgp-5p::gfp activation defect in eft-3(q145);pgp-5p::gfp animals, whereas rme-1 RNAi or rab-5 RNAi did not suppress GFP induction. [Figure 4E] A working model of how the carotenoid extract of K. rhizophylla can suppress the induction of xenobiotic detoxification responses. [Figure 5A] Feeding K. rhizophila inhibited the induction of pgp-5p::gfp in eft-3(q145);pgp-5p::gfp animals, whereas pgp-5p::gfp expression was unaffected in K. rhizophila mutants. [Figure 5B] Feeding K. rhizophila suppressed the induction of pgp-5p::gfp in eft-3(q145);pgp-5p::gfp animals within 12 hours of feeding. [Figure 5C] pgp-5p::gfp animals fed vrs-2 dsRNA and transferred to E. coli showed induction of gfp, whereas animals transferred to K. rhizophila plates had reduced pgp-5p::gfp expression. [Figure 5D] pgp-5p::gfp animals fed with rpl-1 dsRNA and transferred to E. coli showed induction of gfp, whereas animals transferred to K. rhizophila plates had reduced pgp-5p::gfp expression. [Figure 5E]Feeding K. rhizophila significantly reduced the induction of pgp-5p::gfp in eft-3(q145);pgp-5p::gfp animals, whereas pgp-5p::gfp expression was unaffected in K. rhizophila mutants. [Figure 5F] The colony color of the K. rhizophila wild type differed from that of the six mutants. [Figure 5G] Colony colors of K. rhizophila mutants observed from EMS screening. [Figure 6A] K. rhizophila mutants did not suppress the induction of pgp-5p::gfp in eft-3(q145);pgp-5p::gfp animals. [Figure 6B] Observed discoloration phenotypes of 23 mutants whose genomes have been sequenced. [Figure 6C] Discoloration phenotypes observed from C. glutamicum wild type, ΔcrtEb, ΔcrtI, ΔcrtY, and ΔcrtB mutants. [Figure 7-1] Bacterial operon structure containing a putative gene cluster that may produce decaprenoxanthin. [Figure 7-2] This is a continuation of Figure 7-1. [Figure 8-1] Alignment of CrtI proteins from various genera, showing the amino acid conservation across sequences found in the various genera. [Figure 8-2] This is a continuation of Figure 8-1. [Figure 8-3] This is a continuation of Figure 8-2. [Figure 8-4] This is a continuation of Figure 8-3. [Figure 9-1] Alignment of CrtB proteins from various genera, showing the amino acid conservation across sequences found in the various genera. [Figure 9-2] This is a continuation of Figure 9-1. [Figure 10-1] Alignment of CrtEb proteins from various genera, showing the amino acid conservation across sequences found in the various genera. [Figure 10-2]This is a continuation of Figure 10-1. [Figure 11] One exemplary biochemical isolation of a decaprenoxanthin-containing extract from K. rhizophila. [Figure 12A] Spectrophotometric analysis of methanol extracts of K. rhizophila wild type, crtI(e10), and crtb(e6). [Figure 12B] Spectrophotometric analysis of methanol extracts of K. rhizophila wild type, crtEb(e17), and crtYf(e18). [Figure 12C] Spectrophotometric analysis of methanol extracts of K. rhizophila wild type, crtEb(e17), and crtYe(e22). [Figure 13A] K. rhizophylla did not induce hsp-4p::gfp. [Figure 13B] K. rhizophila wild type and crtEb(e17) or crtI(e10) mutants did not induce hsp-6p::gfp. [Figure 13C] K. rhizophila wild type and crtEb(e17), crtI(e10), crtYf(e2), or crtYe(e22) mutants induced clec-60p::gfp. [Figure 13D] K. rhizophila wild type and crtEb(e17) or crtI(e10) mutants did not induce F35E12.5p::gfp. [Figure 13E] The suppression of pgp-5p::gfp induced by feeding K. rhizophila in eft-3(q145);pgp-5p::gfp animals was reversible. [Figure 14A] N-acetylcysteine, ascorbic acid, trolox, or resveratrol did not suppress pgp-5p::gfp in eft-3(q145);pgp-5p::gfp animals. [Figure 14B] Beta-carotene or astaxanthin did not suppress pgp-5p::gfp in eft-3(q145);pgp-5p::gfp animals. [Figure 14C]E. coli expressing either zeaxanthin, neurosporene, violaxanthin, delta-carotene, or alpha-carotene did not suppress pgp-5p::gfp in eft-3(q145);pgp-5p::gfp animals. [Figure 14D] Induction of pgp-5p::gfp in response to 10 μg / ml and 20 μg / ml hygromycin was significantly reduced in animals fed K. rhizophila wild-type. Induction of pgp-5p::gfp in response to 50 mg / ml hygromycin was normal in both E. coli OP50-fed and K. rhizophila wild-type-fed animals. [Figure 15A] Induction of pgp-5p::gfp in response to 2.5 μg / ml or 6.25 μg / ml emetine was significantly reduced in animals fed wild-type K. rhizophila. However, in animals treated with 12.5 μg / ml emetine, induction of pgp-5p::gfp in response was partially reduced in animals fed K. rhizophila. In animals treated with 25 μg / ml emetine, induction of pgp-5p::gfp was normal in both animals treated with either E. coli OP50 or wild-type K. rhizophila. [Figure 15B] Induction of pgp-5p::gfp in response to 6.25 μg / ml emetine was significantly reduced in animals fed K. rhizophila wild type, whereas K. rhizophila crtEb(e17) or K. rhizophila crtI(e10) mutants did not suppress GFP induction. [Figure 15C] Induction of pgp-5p::gfp in response to 1 mM cisplatin was significantly reduced in animals fed K. rhizophila wild type, whereas K. rhizophila crtEb(e17), crtYe(e22), or crtI(e10) mutants did not suppress GFP induction. [Figure 15D] The carotenoid itself was not toxic to the helminths in the absence of hygromycin. [Figure 15E] Animals given the control extract or the K. rhizophila extract were sensitive to antimycin. [Figure 16A]The ZK892.4 and C24A3.4 proteins share sequence similarity. [Figure 16B] Inactivation of ZK892.4 RNAi or C24A3.4 RNAi did not suppress the induction of pgp-5p::gfp in eft-3(q145);pgp-5p::gfp animals, but double RNAi of ZK892.4 and C24A3.4 suppressed gfp induction. [Figure 16C] RNAi of chc-1, fcho-1, or lbp-5 did not induce pgp-5p::gfp. DETAILED DESCRIPTION OF THE INVENTION

[0057] Administration: As used herein, the term "administration" typically refers to the administration of a composition to a subject or system to achieve delivery of an agent to the subject or system. In some embodiments, the agent is a composition or is contained in a composition; in some embodiments, the agent is produced through metabolism of the composition or one or more components thereof. Those skilled in the art will be aware of various routes that, under appropriate circumstances, can be utilized for administration to a subject, e.g., a human. For example, in some embodiments, administration can be ocular, oral, parenteral, topical, etc. In some specific embodiments, administration can be or include one or more of bronchial (e.g., by bronchial instillation), buccal, cutaneous (e.g., topical to the dermis, intradermal, interdermal, transdermal, etc.), enteral, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, intraspecific organ (e.g., intrahepatic), mucosal, intranasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (e.g., by intratracheal instillation), vaginal, vitreous, etc. In many embodiments provided by the present disclosure, administration is oral. In some embodiments, administration can include only a single dose. In some embodiments, administration can require the application of a fixed number of doses. In some embodiments, administration can include dosing that is intermittent (e.g., multiple doses separated by time) and / or periodic (e.g., individual doses separated by a common time period) dosing. In some embodiments, administration can include continuous dosing (eg, perfusion) over at least a selected period of time.

[0058] Analog: As used herein, the term "analog" refers to a substance that shares one or more specific structural features, elements, components, or moieties with a reference substance. Typically, an "analog" exhibits significant structural similarity with the reference substance, e.g., shares a core or consensus structure, but also differs in certain individual ways. In some embodiments, an analog is a substance that can be produced from a reference substance, e.g., by chemical manipulation of the reference substance. In some embodiments, an analog is a substance that can be produced through the implementation of a synthetic process that is substantially similar (e.g., shares multiple steps) to that which produces the reference substance. In some embodiments, an analog is produced, or can be produced, through the implementation of a synthetic process that is different from that used to produce the reference substance.

[0059] Carotenoid Compound: As used herein, the term "carotenoid compound" refers to a compound that is a member of a structurally diverse class of naturally occurring carotenoid pigments and their structural analogs. In nature, carotenoid compounds are typically synthesized from intermediates in the isoprenoid pathway. Carotenoids may be acyclic or cyclic and may or may not contain oxygen; therefore, in some embodiments, the term "carotenoid" can include both carotenes and xanthophylls. Many carotenoids have strong light-absorbing properties. Generally, carotenoids are hydrocarbon compounds with a conjugated polyene carbon skeleton formally derived from the five-carbon compound isopentenyl pyrophosphate. In some embodiments, carotenoid compounds can be triterpenes (C30 diapocarotenoids), tetraterpenes (C40 carotenoids), or other compounds that are, for example, C35, C50, C60, C70, C80, or other lengths. In some embodiments, the carotenoid may have a length greater than C200. Over 1000 different carotenoids have been identified in nature.Carotenoids include, but are not limited to, antheraxanthin, adonirubin, adonixanthin, astaxanthin, canthaxanthin, capsorubrin, β-cryptoxanthin, α-carotene, β-carotene, β,ψ-carotene, δ-carotene, ε-carotene, echinenone, 3-hydroxyechinenone, 3'-hydroxyechinenone, γ-carotene, ψ-carotene, 4-keto-γ-carotene, ζ-carotene, α-cryptoxanthin, deoxyflexixanthin, diatoxanthin, 7,8-didehydroataxanthin, didehydrolycopene, and fucoxanthin. Carotenoids include fucoxanthinol, isorenieratene, β-isorenieratene, lactucaxanthin, lutein, lycopene, myxobactone, neoxanthin, neurosporene, hydroxyneurosporene, peridinin, phytoene, rhodopin, rhodopin glucoside, 4-keto-rubicxanthin, siphonaxanthin, spheroidene, spheroidenone, spirilloxanthin, tolulene, 4-keto-tolulene, 3-hydroxy-4-keto-tolulene, uriolide, uriolide acetate, violaxanthin, zeaxanthin-β-diglucoside, zeaxanthin, and C30 carotenoids. Additionally, carotenoid compounds include derivatives of these molecules that contain hydroxy-, methoxy-, oxo-, epoxy-, carboxy-, or aldehyde functional groups. For example, carotenoids include oxygenated derivatives. Additionally, included carotenoid compounds include esters (e.g., glycoside esters, fatty acid esters) and sulfate derivatives (e.g., esterified xanthophylls).

[0060] Modified carotenogenesis: As used herein, the term "modified carotenogenesis" refers to a modification of a host organism that regulates the production of one or more carotenoids as described herein. For example, modified carotenogenesis can increase the production level of one or more carotenoids and / or change the relative production levels of different carotenoids. In principle, modified carotenogenesis of the invention can be any chemical, physiological, genetic, or other modification that appropriately alters the production of one or more carotenoids in a host organism produced by the organism when compared to the levels produced in an otherwise identical organism not subjected to the same modification. However, in most embodiments, modified carotenogenesis will typically involve a genetic modification that results in increased production of one or more selected carotenoids. In some embodiments, the selected carotenoids are one or more C50 carotenoid compounds.

[0061] Carotenoid biosynthetic polypeptide: The term "carotenoid biosynthetic polypeptide" refers to any polypeptide involved in the synthesis of one or more carotenoids. These carotenoid biosynthetic polypeptides include, for example, phytoene synthase, phytoene dehydrogenase (or desaturase), lycopene cyclase, carotenoid ketolase, carotenoid hydroxylase, astaxanthin synthase, carotenoid epsilon hydroxylase, lycopene cyclase (beta and epsilon subunits), carotenoid glucosyltransferase, and acyl-CoA:diacylglycerol acyltransferase polypeptides, to name a few.

[0062] Microorganisms that synthesize carotenoid compounds: As used herein, the phrase "microorganisms that synthesize carotenoid compounds" refers to microorganisms (e.g., algae, fungi, bacteria) that synthesize one or more carotenoid compounds. In some embodiments, the microorganisms that synthesize carotenoid compounds can naturally synthesize one or more carotenoid compounds. In some embodiments, the microorganisms that synthesize carotenoid compounds include modifications of carotenogenesis. In some embodiments, the microorganisms that synthesize carotenoid compounds may be genetically modified (e.g., to have one or more gene mutations) so that they synthesize one or more carotenoids at absolute or relative levels that differ from those of an otherwise equivalent reference microorganism that has not been genetically modified (i.e., does not contain gene mutations). For example, in some embodiments, the microorganisms that synthesize carotenoid compounds are genetically engineered to synthesize at least one carotenoid compound that is not synthesized by a microorganism without genetic engineering. Alternatively, in some embodiments, the microorganism that synthesizes carotenoid compounds is genetically engineered so that its synthesis of one or more specific carotenoid compounds can be at a higher level compared to a microorganism without genetic engineering. In some embodiments, the higher level can be assessed with reference to a threshold level; in some embodiments, the higher level can be assessed with reference to another compound (e.g., another carotenoid compound) that is also produced by the microorganism (prior to genetic engineering). In some specific embodiments, the microorganism that synthesizes carotenoid compounds is genetically engineered to add or increase the expression of one or more genes encoding carotenoid biosynthetic polypeptides. Alternatively, or in addition, in some embodiments, the microorganism that synthesizes carotenoid compounds is genetically engineered to increase carbon flow through the carotenoid biosynthetic pathway (e.g., by reducing carbon diversion to one or more other biosynthetic or metabolic pathways). In some embodiments, the microorganism that synthesizes carotenoid compounds can synthesize one or more carotenoid compounds with a specific number of carbon units.For example, in some embodiments, a microorganism that synthesizes carotenoid compounds can synthesize one or more C50 carotenoid compounds (naturally or as a result of genetic modification); such a microorganism can be referred to as a C50-synthesizing microorganism.

[0063] Equivalent: As used herein, the term "equivalent" refers to two or more agents, entities, circumstances, sets of conditions, etc. that may not be identical to one another, but are similar enough to permit comparisons between them so that a person of ordinary skill in the art would recognize that conclusions can be reasonably drawn based on observed differences or similarities. In some embodiments, equivalent sets of conditions, environments, individuals, or populations are characterized by multiple substantially identical characteristics and one or a few different characteristics. A person of ordinary skill in the art will understand in context the degree of identity required in any given environment for two or more such agents, entities, circumstances, sets of conditions, etc. to be considered equivalent. For example, a person of ordinary skill in the art will recognize that sets of environments, individuals, or populations are equivalent to one another when they are characterized by a sufficient number and type of substantially identical characteristics to warrant a reasonable conclusion that differences in results obtained or phenomena observed under or with different sets of environments, individuals, or populations are caused by, or indicate, variations in those characteristics.

[0064] Dosage form: Those of skill in the art will recognize that the term "dosage form" can be used to refer to a physically discrete unit of an agent (e.g., a therapeutic agent) for administration to a subject. Typically, each such unit contains a predetermined quantity of the agent. In some embodiments, such quantity is a unit dosage (or a whole fraction thereof) that is appropriate for administration according to a dosing regimen (i.e., a therapeutic dosing regimen) determined to correlate with a desired or beneficial outcome when administered to a relevant population. Those of skill in the art will recognize that the total amount of a therapeutic composition or agent to be administered to a particular subject is determined by one or more attending physicians and can include the administration of multiple dosage forms.

[0065] Dosage regimen: Those skilled in the art will recognize that the term "dosage regimen" can be used to refer to a series of unit doses (typically more than one) administered individually to a subject, typically separated by a time period. In some embodiments, a given agent has a recommended dosing regimen, and the dosing regimen can include one or more doses. In some embodiments, the dosing regimen includes multiple doses, each separated temporally from the other doses. In some embodiments, the individual doses are separated from each other by periods of equal length; in some embodiments, the dosing regimen includes multiple doses and at least two different periods separating the individual doses. In some embodiments, all doses within the dosing regimen are of the same unit dosage. In some embodiments, different doses within the dosing regimen are of different amounts. In some embodiments, the dosing regimen includes a first dose at a first dosage amount, followed by one or more additional doses at a second dosage amount that is different from the first dosage amount. In some embodiments, the dosing regimen includes a first dose at a first dosage amount, followed by one or more additional doses at a second dosage amount that is the same as the first dosage amount. In some embodiments, the dosing regimen correlates with a desirable or beneficial outcome when administered across a relevant population.

[0066] Engineered: Generally, the term "engineered" refers to aspects of manipulation by human hands. For example, a cell or organism is considered "engineered" if it has been manipulated in such a way that its genetic information is altered (e.g., new genetic material not previously present is introduced, such as by transformation, mating, somatic hybridization, transfection, transduction, or other mechanisms, or previously present genetic material is altered or removed, for example, by substitution or deletion mutations or by mating protocols). As is common and understood by those of skill in the art, the progeny of an engineered polynucleotide or cell are typically still referred to as "engineered," despite the actual manipulation that occurred on the previous entity.

[0067] Excipient: As used herein, refers to an inactive (e.g., non-therapeutic) agent that can be included in a pharmaceutical composition to provide or contribute, for example, to a desired consistency or stabilizing effect. In some embodiments, suitable pharmaceutical excipients can include, for example, starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, nonfat dry milk, glycerol, propylene, glycol, water, ethanol, etc.

[0068] Functional: As used herein, a "functional" biological molecule is a biological molecule in a form in which it exhibits the properties and / or activities by which it is characterized. A biological molecule can have two functions (i.e., bifunctional) or many functions (i.e., multifunctional).

[0069] Enhance, increase, inhibit, or reduce: As used herein, the terms "enhance," "increase," "inhibit," "reduce," or their grammatical equivalents refer to a value compared to a baseline or other reference measurement. In some embodiments, a suitable reference measurement may be or include a measurement in a particular system (e.g., in a single individual) under otherwise equivalent conditions in the absence of a particular agent or treatment (e.g., before and / or after a particular agent or treatment), or in the presence of an appropriate equivalent reference agent. In some embodiments, a suitable reference measurement may be or include a measurement in an equivalent system known or expected to respond in a particular way in the presence of the relevant agent or treatment. In some embodiments, a suitable reference is a negative reference; in some embodiments, a suitable reference is a positive reference.

[0070] Isolated: As used herein, this refers to a substance and / or entity that (1) has been separated from at least some of the components with which it was associated when originally created (whether in nature and / or in an experimental setting) and / or (2) has been designed, created, prepared, and / or manufactured by the hand of man. In some embodiments, an isolated substance or entity may be enriched; in some embodiments, an isolated substance or entity may be pure. In some embodiments, an isolated substance and / or entity may be separated from about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% of the other components with which it was originally associated. In some embodiments, an isolated agent is about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% pure. As used herein, a substance is "pure" when it is substantially free of other components. In some embodiments, as those skilled in the art will understand, a substance can still be considered "enriched," "isolated," or even "pure" even after being combined with certain other components, such as, for example, one or more carriers or excipients (e.g., buffers, solvents, water, etc.); in such embodiments, the isolation or purity percentage of a substance is calculated without including such carriers or excipients. Those skilled in the art are aware of various techniques for isolating (e.g., enriching or purifying) a substance or agent (e.g., using one or more of fractionation, extraction, precipitation, or other separations).

[0071] Pharmaceutical composition: As used herein, the term "pharmaceutical composition" refers to a composition in which an active agent is formulated with one or more pharmaceutically acceptable carriers. In some embodiments, the active agent is present in a unit dosage suitable for administration in a treatment regimen that exhibits a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population. In some embodiments, pharmaceutical compositions can be specially formulated for administration in solid or liquid form, including those suitable for oral administration, e.g., drenches (aqueous or non-aqueous solutions or suspensions), tablets, e.g., those intended for buccal, sublingual, and systemic absorption, boluses, powders, granules, pastes for application to the tongue, capsules, powders, etc. In some embodiments, the active agent can be or include a cell or a population of cells (e.g., a culture of a microorganism that synthesizes a carotenoid compound); in some embodiments, the active agent can be or include an extract or component of a cell or a population of cells (e.g., a culture). In some embodiments, the active agent can be or comprise an isolated, purified, or pure compound. In some embodiments, the active agent may be synthesized in vitro (e.g., by chemical and / or enzymatic synthesis). In some embodiments, the active agent can be or comprise a natural product (whether isolated from its natural source or synthesized in vitro).

[0072] Pharmaceutically acceptable: As used herein, the term "pharmaceutically acceptable," which may be used in reference to a carrier, diluent, or excipient used, for example, to formulate a pharmaceutical composition disclosed herein, means the carrier, diluent, or excipient must be compatible with the other ingredients of the composition and not deleterious to the recipient thereof.

[0073] Pharmaceutically acceptable carrier: As used herein, the term "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, or solvent encapsulant, that is involved in the transport or transfer of a compound of interest from one organ or part of the body to another. Each carrier should be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include: sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethyl cellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; pH buffer solutions; polyesters, polycarbonates, and / or polyanhydrides; and other non-toxic, compatible substances used in pharmaceutical formulations.

[0074] Prevention: As used herein, the term "prevention" refers to delaying the onset of one or more symptoms of a particular disease, disorder, or condition, and / or reducing the frequency and / or severity of said symptoms. In some embodiments, prevention is assessed on a population basis; thus, an agent is considered to "prevent" a particular disease, disorder, or condition if a statistically significant reduction in the occurrence, frequency, and / or intensity of one or more symptoms of the disease, disorder, or condition is observed in a population susceptible to the disease, disorder, or condition. In some embodiments, prevention can be considered complete, for example, if the onset of the disease, disorder, or condition is delayed for a predetermined period of time.

[0075] Reference: As used herein, this describes a standard or control against which a comparison is made. For example, in some embodiments, an agent, animal, individual, population, sample, sequence, or value of interest is compared to a reference or control agent, animal, individual, population, sample, sequence, or value. In some embodiments, the reference or control is tested and / or determined substantially simultaneously with the test or determination of interest. In some embodiments, the reference or control is a historical reference or control, optionally embodied in a tangible medium. Typically, as would be understood by those skilled in the art, the reference or control is determined or characterized under conditions or circumstances equivalent to those under evaluation. Those skilled in the art will recognize that sufficient similarity justifies reliance on and / or comparison with a particular possible reference or control. In some embodiments, the reference is a negative control reference; in some embodiments, the reference is a positive control reference.

[0076] Risk: As understood from the context, "risk" of a disease, disorder, and / or condition refers to the likelihood that a particular individual will develop the disease, disorder, and / or condition. In some embodiments, risk is expressed as a percentage. In some embodiments, risk is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90 to 100%. In some embodiments, risk is expressed as risk compared to the risk associated with a reference sample or a group of reference samples. In some embodiments, the reference sample or a group of reference samples has a known risk of the disease, disorder, condition, and / or event. In some embodiments, the reference sample or a group of reference samples is derived from an individual comparable to the particular individual. In some embodiments, the relative risk is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more.

[0077] Sample: As used herein, the term "sample" typically refers to an aliquot of material obtained from or derived from a source of interest. In some embodiments, the source of interest is a biological or environmental source. In some embodiments, the source of interest can be or include a cell or organism, such as a microorganism, a plant, or an animal (e.g., a human). In some embodiments, the source of interest is or includes a biological tissue or fluid. In some embodiments, the biological tissue or fluid may be or include amniotic fluid, aqueous humor, ascites, bile, bone marrow, blood, milk, cerebrospinal fluid, earwax, chyle, chime, ejaculate, endolymph, exudate, feces, gastric acid, gastric juice, lymph, mucus, pericardial fluid, perilymph, peritoneal fluid, pleural fluid, pus, mucosal secretions, saliva, sebum, semen, serum, smegma, sputum, synovial fluid, sweat, tears, urine, vaginal secretions, vitreous humor, vomit, and / or combinations or components thereof. In some embodiments, the biological fluid may be or include intracellular fluid, extracellular fluid, intravascular fluid (plasma), interstitial fluid, lymph, and / or pericellular fluid. In some embodiments, the biological fluid may be or include plant exudates. In some embodiments, the biological tissue or sample can be obtained, for example, by aspiration, biopsy (e.g., fine needle or tissue biopsy), swab (e.g., oral, nasal, skin, or vaginal swab), scraping, surgery, washing, or irrigation (e.g., bronchoalveolar epithelium, duct, nasal, ocular, oral, uterine, vaginal, or other washing or irrigation). In some embodiments, the biological sample is or includes cells obtained from an individual. In some embodiments, the sample is a "primary sample" obtained directly from the source of interest by any suitable means. In some embodiments, as is clear from the context, the term "sample" refers to a preparation obtained by processing the primary sample (e.g., by removing one or more components of the primary sample and / or adding one or more agents to the primary sample). For example, filtration using a semipermeable membrane.Such a "processed sample" can include, for example, nucleic acids or proteins extracted from a sample or obtained by subjecting the primary sample to one or more techniques, such as amplification or reverse transcription of nucleic acids, isolation and / or purification of certain components, etc.

[0078] Subject: As used herein, the term "subject" refers to an individual to whom the provided treatment is administered. In some embodiments, the subject is a mammal, such as a mammal experiencing or susceptible to a disease, disorder, or condition described herein; in some embodiments, the subject is a human or non-human veterinary subject, such as an ape, cat, dog, monkey, or pig. In some embodiments, the subject is a human. In some embodiments, the patient is suffering from or susceptible to one or more diseases, disorders, or conditions described herein. In some embodiments, the patient exhibits one or more symptoms of one or more diseases, disorders, or conditions described herein. In some embodiments, the patient has been diagnosed with one or more diseases, disorders, or conditions described herein. In some embodiments, the disorder or condition is or includes nausea and / or vomiting and / or one or more food aversion disorders. In some embodiments, the subject is suffering from or susceptible to cancer or the presence of one or more tumors. In some embodiments, the subject is undergoing or has undergone a particular therapy to diagnose and / or treat a disease, disorder, or condition, hi some embodiments, the subject has undergone a therapy (e.g., chemotherapy, radiation, and / or surgery) that induces nausea and / or vomiting.

[0079] Symptoms are reduced: According to the present invention, a "symptom is reduced" when the magnitude (e.g., intensity, severity, etc.) and / or frequency of one or more symptoms of a particular disease, disorder, or condition is reduced. For purposes of clarity, delaying the onset of a particular symptom is considered a form of reducing the frequency of that symptom.

[0080] Therapeutic regimen: "Therapeutic regimen," as that term is used herein, refers to a dosing regimen that, when administered throughout a relevant population, can be correlated with a desirable or beneficial therapeutic outcome.

[0081] Therapeutically effective amount: As used herein, this refers to an amount that produces a desired effect in those to whom it is administered. In some embodiments, the term refers to an amount sufficient to treat a disease, disorder, and / or condition when administered to a population suffering from or susceptible to the disease, disorder, and / or condition according to a therapeutic dosing regimen. In some embodiments, a therapeutically effective amount is an amount that reduces the incidence and / or severity of one or more symptoms of the disease, disorder, and / or condition and / or delays the onset of the symptoms. Those skilled in the art will recognize that the term "therapeutically effective amount" does not actually require that a successful treatment be achieved in a particular individual. Rather, a therapeutically effective amount may be such an amount that, when administered to patients in need of such treatment, provides a specific, desired pharmacological response in a significant number of subjects. In some embodiments, reference to a therapeutically effective amount may be a reference to an amount measured in one or more specific tissues (e.g., tissues affected by a disease, disorder, or condition) or fluids (e.g., blood, saliva, serum, sweat, tears, urine, etc.). Those skilled in the art will recognize that in some embodiments, a therapeutically effective amount of a particular agent or therapy can be formulated and / or administered in a single dose, hi some embodiments, a therapeutically effective agent can be formulated and / or administered in multiple doses, e.g., as part of a dosing regimen.

[0082] Treatment: As used herein, the term "treatment" (also "treat" or "treating") refers to any application of therapy that partially or completely alleviates, improves, alleviates, inhibits, delays the onset of, reduces the severity of, and / or reduces the incidence of one or more symptoms, characteristics, and / or causes of a particular disease, disorder, and / or condition. In some embodiments, such treatment may be of subjects who do not exhibit symptoms of the associated disease, disorder, and / or condition and / or who exhibit only early signs of the disease, disorder, and / or condition. Alternatively, or in addition, such treatment may be of subjects who exhibit symptoms of one or more established associated diseases, disorders, and / or conditions. In some embodiments, treatment may be of subjects who have been diagnosed with the associated disease, disorder, and / or condition. In some embodiments, treatment may be of subjects who are known to have one or more susceptibility factors that are statistically correlated with an increased risk of developing the associated disease, disorder, and / or condition.

[0083] Detailed Description of Certain Embodiments The nematode Caenorhabditis elegans monitors defects in essential cellular activities, such as those caused by microbial toxins, and responds by activating the cellular surveillance-activated detoxification and defenses (cSADD) response (Melo and Ruvkun, 2012). Ribosomal RNA and protein abundance, as well as other proteins that mediate mRNA translation into proteins, can be targets of microbial toxins and virulence factors. The activity of these translational components can be monitored to detect attenuation of protein synthesis caused by toxins or mutations. Detection of attenuation is coupled via signaling cascades to activate xenobiotic detoxification and behavioral responses, such as food aversion, via p38 MAPK, bZIP / ZIP-2 transcription factors, and the bile acid biosynthesis pathway (Melo and Ruvkun, 2012; Dunbar et al., 2012; Govindan et al., 2015). By monitoring the attenuation of core cellular functions rather than the molecular structure of unknown toxins, C. elegans can detect unexpected pathogens and toxins. Many components of this signaling cascade, such as MAP kinases and the bile acid biosynthetic pathway, are conserved across animals; this disclosure recognizes that the cSADD system of toxin monitoring and response can be applied to animals other than C. elegans and further teaches that C. elegans may be useful as a model system for characterizing agents that modulate this system, and may be useful in certain therapeutic applications described herein.

[0084] The present disclosure further recognizes that animal defense strategies similar or homologous to the C. elegans cSADD system may drive the evolution of bacterial countermeasures to thwart these defense responses; there are many examples of such evolutionary competition between hosts and pathogens. The present disclosure also recognizes that commensal bacteria may attempt to quell such animal defense responses in order to establish a favorable or symbiotic relationship.

[0085] The present disclosure further recognizes that microorganisms (e.g., bacteria) synthesize a variety of chemical compounds with significant biological activity, including compounds that target ribosomes and / or associated translation factors (Berdy et al., 2005); such microorganisms have proven to be productive sources of drugs or drug candidate compounds. The present disclosure (i) teaches useful therapeutic applications of agents that can inhibit certain detoxification pathways; (ii) provides a system for evaluating one or more properties of an agent relative to the inhibition of such detoxification pathways; and (iii) identifies potential sources of such agents (e.g., microorganisms such as bacteria, which in some embodiments may be commensal microorganisms).

[0086] For example, the present disclosure provides that certain bacterial strains (e.g., those that produce certain carotenoid compounds, particularly certain C50 carotenoid compounds) may be excellent sources of agents (e.g., certain useful carotenoid compounds) useful according to the present disclosure. Additionally, the present disclosure provides techniques for assessing one or more relevant properties of such agents.

[0087] In a microbial inhibitory screen of diverse bacteria for activity in suppressing the activation of xenobiotic detoxification genes in C. elegans with a genetically induced translation defect, potent activity was identified from wild-type K. rhizophila (Govindan et al., 2015). Using genetic analysis of K. rhizophila, we demonstrated that C50 carotenoid compounds can suppress the translational toxin defense response of C. elegans.

[0088] As shown herein, K. rhizophila, which produces the C50 carotenoid compound decaprenoxanthin, inhibited cSADD translation and the DNA-damaging toxin defense response in C. elegans. Mutants of the K. rhizophila C50 carotenoid biosynthetic pathway did not inhibit this xenobiotic detoxification response in C. elegans (Figure 1A-C). K. rhizophila extract suppressed this xenobiotic detoxification response in C. elegans and further restored the ability of K. rhizophila carotenoid mutants to achieve such suppression (Figure 2B & C). K. rhizophila extract also suppressed the induction of the C. elegans xenobiotic detoxification pathway in response to translation-targeting toxins (Figure 3A, Figure 1C). K. rhizophila C50 carotenoid compounds also suppressed the response to RNA interference inactivation of other genes required for protein synthesis, such as core ribosomal proteins or aminoacyl-tRNA synthases. C50 carotenoids from C. glutamicum also inhibited the induction of the xenobiotic detoxification pathway in C. elegans in response to translational defects, indicating that C50 carotenoid regulation of C. elegans surveillance was not limited to one particular bacterial lineage.

[0089] The present disclosure teaches that certain carotenoid compounds (e.g., certain C50 carotenoid compounds) are useful for a variety of therapeutic applications, including, inter alia, the treatment of nausea and / or vomiting (particularly including induced nausea and / or vomiting, such as chemotherapy-induced nausea and / or vomiting). Carotenoids have been most studied in photosynthetic bacteria and plants, where carotenoids are auxiliary light-absorbing components of photosynthesis (Edge et al., 1997). In photosynthetic chlorophyll clusters, carotenoids absorb light energy and transfer it to the photosynthetic electron transport chain, where the reduction potential energy of photoexcited electrons then triggers the physical movement of multiple iron-sulfur and heme proteins to move protons across the lipid bilayer, generating a pH gradient. Common to photosynthetic bacteria is the strong light absorption and lipid solubility of these pigments at visible light wavelengths. These bacteria are highly pigmented because carotenoids are highly abundant and possess a conjugated double bond system that delocalizes electrons into an unusually large, resonantly stabilized potential well, with orbital transitions at energies much lower than those of standard biochemical bonds. Carotenoids are also known antioxidants in photosynthesis. However, as shown herein, the antioxidant activity of carotenoids does not explain the suppression of drug detoxification responses in animals, as other antioxidants were unable to suppress surveillance in C. elegans.

[0090] C50 carotenoid compounds can affect animal monitoring of translational components through simple changes in membrane fluidity. As shown herein, C50 carotenoid compounds suppressed the induction of xenobiotic detoxification responses in C. elegans by inhibiting the bile acid biosynthesis pathway. While bile acids were traditionally thought of as fat emulsifiers that aid digestion, several recent studies have discovered that bile acids are important signaling molecules in metabolic and immune pathways. Bile acid signaling may be involved in the induction of CINV or drug detoxification responses in humans. The lipid solubility and abundance of carotenoids may contribute to their anti-bile acid signaling function in C. elegans monitoring of translation and DNA damage responses.

[0091] The cSADD response can not only induce xenobiotic detoxification but also induce food aversion behavior. Food aversion is a relevant animal response because many toxins are derived from bacterial pathogens that can be associated with or cause food spoilage. The induction of xenobiotic detoxification and food aversion associated with bacterial immune pathways may be animal programs derived from this evolutionary history. This disclosure provides insights into the possibility that chemotherapy-induced nausea and vomiting (CINV) responses in humans may be associated with these xenobiotic aversion programs. Interestingly, cisplatin, which is used to block DNA replication in cancer patients, has a high emetogenic potential. Emetine, an antibiotic that targets eukaryotic protein synthesis, is also highly emetogenic, as its name suggests. These two drugs were able to induce not only a xenobiotic detoxification response in C. elegans but also induced strong food aversion. The pgp-5 ABC transporter gene was strongly induced by toxins that cause DNA damage and even by chemically distinct toxins that inhibit translation (but not by a wide range of toxins that target other pathways, such as mitochondria or the ER) (Govindan et al., 2015). Thus, the present disclosure teaches that C. elegans food aversion is an excellent model for studying the mechanisms underlying human CINV.

[0092] The serotonergic pathway is involved in both food aversion in C. elegans and CINV in humans (Melo and Ruvkun, 2012). Hepatic drug detoxification and elimination are key concerns in cancer chemotherapy; amplification of ABC transporters, which expedite drug elimination, is frequently observed in drug-resistant cancer patients. K. rhizophila carotenoid extracts induced hypersensitivity to xenobiotics that target protein translation and cause DNA damage by suppressing the induction of drug detoxification pathways. Furthermore, C50 carotenoid compounds from K. rhizophila suppressed food aversion induced by the emetic toxins emetine and cisplatin.

[0093] C50 carotenoid compounds, such as decaprenoxanthin, C50-astaxanthin, C50-β-carotene, C50-carotene (n=3) (16,16-diisopentenylphytoene), C50-zeaxanthin, C50-caloxanthin, C50-nostoxanthin, sarcinaxanthin, sarprenoxanthin, the acyclic C50 carotenoid bacterioruberin, C50-canthaxanthin, C50-lycopene, C50-phytoene, can be used in accordance with the present disclosure as therapies for treating and / or reducing the risk of nausea and / or vomiting, e.g., induced nausea and / or vomiting, e.g., CINV, RINV, etc. Alternatively, or in addition, the present disclosure provides that such C50 carotenoid compounds may be useful as therapies for treating and / or reducing the risk of one or more food aversion disorders (e.g., anorexia nervosa). Furthermore, the present disclosure provides a system for evaluating one or more properties of an agent related to its usefulness as a therapy described herein (i.e., for treating and / or reducing the risk of nausea and / or vomiting and / or one or more food aversion disorders), and further demonstrates that such useful agents include compounds produced by certain microorganisms (e.g., bacteria), including certain commensal microorganisms. Using such a system, the present disclosure describes the usefulness of certain carotenoid compounds (e.g., C50 carotenoid compounds), including those produced by various bacterial strains (e.g., synthesized by bacterial enzymes). Those skilled in the art will recognize, upon reading this disclosure, that a variety of other compounds (including a variety of other carotenoid compounds) can be (and / or have been) produced by microorganisms and / or produced by chemical synthesis and will be evaluated for activities such as those embodied in the systems exemplified therein.Thus, one of skill in the art will recognize, upon reading this disclosure, that various chemical agents, including carotenoid compounds, specifically exemplified by C50 carotenoids, are provided that are suitable for the evaluations described herein and / or useful as therapeutic agents described herein.

[0094] Treatment method The methods provided by the present disclosure include methods for treating certain diseases, disorders and conditions.In some embodiments, the related diseases, disorders and conditions can be or include nausea and / or vomiting and / or certain food aversion disorders.In some embodiments, the nausea and / or vomiting that can be treated as described herein can be associated with one or more of motion sickness, seasickness, pregnancy (e.g., morning sickness or hyperemesis gravidarum), pain, emotional stress, gallbladder disease, heart attack, concussion or brain injury (e.g., brain tumor), overeating, gallbladder disease, infection, ulcer, gastroparesis, intestinal obstruction, appendicitis, infection (e.g., viral infection), etc.

[0095] In some embodiments, the nausea and / or vomiting that can be treated as described herein can be, for example, induced nausea and / or vomiting, such as induced by ingestion of a toxin or other exposure to a toxin (e.g., food poisoning, drug-induced nausea and / or vomiting, intoxication, etc.). In some embodiments, the induced nausea and / or vomiting that can be treated as described herein can be nausea and / or vomiting associated with chemotherapy or radiation. In some embodiments, the induced nausea and / or vomiting that can be treated as described herein can be CINV or radiation-induced nausea and vomiting (RINV); at least three types of emesis, namely, acute emesis, delayed emesis, and anticipatory emesis, are generally associated with the use of chemotherapy agents. In some embodiments, the induced nausea and / or vomiting that can be treated as described herein can be or include, for example, postoperative nausea and vomiting (PONV).

[0096] In general, the methods of treatment provided by the present disclosure involve administering to a subject in need of, or determined to be in need of, such treatment a therapeutically effective amount of a carotenoid compound described herein.

[0097] In some embodiments, the methods of treatment provided herein are prophylactic or preventative, e.g., can be administered to a subject prior to the onset of significant symptoms and / or prior to exposure to a particular expected trigger associated with nausea and / or vomiting (e.g., chemotherapy, radiation therapy, surgery, or other treatment (e.g., pharmacological treatment) associated with nausea and / or vomiting).

[0098] In some embodiments, the methods of treatment provided herein are therapeutic, e.g., can be administered to a subject after the onset of significant symptoms of nausea and / or vomiting (e.g., during or after at least one episode of nausea or vomiting).

[0099] In preferred embodiments, the provided methods of treatment are administered to a subject that is a mammal, e.g., a mammal experiencing a disease, disorder, or condition described herein; in some embodiments, the subject is a human or non-human veterinary subject, e.g., an ape, cat, dog, monkey, or pig.

[0100] In many embodiments, "treatment" includes improving at least one symptom of a disease, disorder, or condition associated with nausea. Often, nausea leads to food aversion, loss of appetite, and / or reduced calorie intake, and potentially weight loss; in some embodiments, administration of a therapeutically effective amount of a carotenoid compound described herein can result in reduced nausea, vomiting, and / or food aversion. Alternatively, or in addition, in some embodiments, administration of a therapeutically effective amount of a carotenoid compound described herein can achieve a restoration of appetite and / or a return to or approach to normal calorie intake, a reduction, cessation, or slowing of weight loss, an increase in weight / weight gain, and / or a reduction in the frequency, duration, or severity of current or future episodes of nausea, vomiting, food aversion, and / or loss of appetite.

[0101] In some embodiments, the methods can include administering a therapeutically effective amount of a carotenoid compound to a subject about to undergo, for example, chemotherapy, radiation therapy, or other treatment associated with nausea and vomiting, prior to, during (e.g., simultaneously with), or after administration of the treatment expected to be associated with nausea and / or vomiting.

[0102] In some embodiments, subjects receiving the treatments described herein may be undergoing and / or may have undergone other treatments (e.g., chemotherapeutic, radiotherapeutic, surgical, etc.) that may, for example, induce vomiting or that may be intended to treat one or more symptoms or characteristics of a disease, disorder, or condition described herein, and thus the carotenoid therapy provided is administered in combination with such other therapies to treat the relevant disease, disorder, or condition.

[0103] Carotenoid Composition Among other things, the present disclosure provides compositions that include or otherwise deliver carotenoid compounds (e.g., C50 carotenoid compounds) to a subject suffering from or susceptible to one or more diseases, disorders, or conditions described herein.

[0104] Those skilled in the art are aware that over 750 carotenoid compounds have previously been identified. Carotenoids are generally classified by the number of five-carbon isoprenoid units that give them carbon skeletons of various lengths. For example, Henke et al., (2017). C50 Carotenoids: Occurrence, Biosynthesis, Glycosylation, and Metabolic Engineering for their Overproduction. In, Bio-pigmentation and Biotechnological Implementations, pp.107-126. doi:10.1002 / 9781119166191.ch5;Fernandes, Introductory Chapter: Carotenoids - A Brief Overview on Its Structure, Biosynthesis, Synthesis, and Applications. In Progress in Carotenoid Research. (2018) doi:10.5772 / intechopen.79542;Mezzomo et al., (2016) Carotenoids Functionality, Sources, and Processing by Supercritical Technology: A Review in J. Chemistry Volume Please refer to 2016:1.

[0105] In some embodiments, carotenoid compounds useful according to the present disclosure have a relatively long isoprene backbone, for example, having a length in the range of about 45 to about 60 carbon atoms. In some embodiments, useful carotenoid compounds described herein may have an isoprene backbone that is 50 carbons long, i.e., a C50 carotenoid compound, such as decaprenoxanthin, or other C50 carotenoids, such as C50-astaxanthin (also called decaprenoastaxanthin; see, e.g., Milon et al., Helv. Chim. Acta 69, 12-24 (1986); Furubayashi et al., Nat Commun. 2015; 6: 7534) or C50-β-carotene (also called decapreno-β-carotene) (see, e.g., Karrer et al., Helv. Chim. Acta 34, 28-33 (1951); Furubayashi et al., Nat Commun. 2015; 6: 7534); 16,16'-diisopentenylphytoene (Umeno et al., J Bacteriol, 186, 1531-1536 (2004); C50-carotene (n=3) (16,16-diisopentenylphytoene), C50-zeaxanthin, C50-caloxanthin, and C50-nostoxanthin (US20140170700); sarcinaxanthin, sarprenoxanthin, the acyclic C50 carotenoid bacterioruberin, C50-canthaxanthin, C50-lycopene, and C50-phytoene (Li et al., Scientific Reports volume 9, Article number: 2982 (2019)). See also Pfander, Pure and Applied Chemistry, 66(10-11):2369-2374 (1994).

[0106] Those skilled in the art are aware of various techniques for producing carotenoid compounds. See, for example, Mezzomo et al., (2016) Carotenoids Functionality, Sources, and Processing by Supercritical Technology: A Review in J. Chemistry Volume 2016:1. In some embodiments, carotenoid compounds can be isolated from the organisms (e.g., plants or microorganisms) that produced them. In some such embodiments, such plants or microorganisms have been developed and / or cultivated by humans. In some embodiments, the plants or microorganisms may be naturally occurring plants or microorganisms. In some embodiments, the plants or microorganisms may be engineered plants or microorganisms (e.g., plants or microorganisms engineered to synthesize the carotenoid compounds described herein).

[0107] Those skilled in the art are aware of a variety of plant and / or microbial sources that have been or can be engineered to produce plants or microorganisms that synthesize the carotenoid compounds described herein. See, e.g., WO2016 / 102342.

[0108] In some embodiments, carotenoid compounds can be isolated from plant or microbial sources (e.g., from cultures or cultures thereof). Those skilled in the art will know various techniques for processing plant and / or microbial cells or tissues, for example, to prepare extracts thereof and / or to isolate components and / or compounds therefrom.

[0109] Alternatively, or in addition, in some embodiments, carotenoid compounds can be prepared partially or completely in vitro (e.g., by chemical and / or enzymatic synthesis, or a combination thereof), and optionally further isolated and / or purified as known in the art.

[0110] A variety of techniques are known in the art that can be used to prepare extracts of cells or organisms that produce the relevant carotenoid compounds and / or to isolate extracts, components, or compounds therefrom, or to process in vitro carotenoid synthesis systems (e.g., to isolate and / or purify one or more carotenoid compounds from an in vitro carotenoid synthesis system). Such techniques can include, for example, one or more of organic extraction, vacuum concentration, chromatography, etc., to name just a few.

[0111] Those skilled in the art know that various lower (shorter carbon chain) carotenoids, such as α-carotene, β-carotene, γ-carotene, δ-carotene, ε-carotene, lutein, zeaxanthin, canthaxanthin, fucoxanthin, astaxanthin, antheraxanthin, and violaxanthin, are synthesized by terminal modification of lycopene by cyclization or oxidation. Higher (more carbon atoms) carotenoids can be prepared, for example, using in vitro methods or in vivo or ex vivo methods (e.g., via natural or genetically modified organisms). For example, C50 carotenoids can be synthesized in vitro by adding two dimethylallyl pyrophosphate (DMAPP) molecules to the C(2) and C(2') of each C40 carotenoid, or can be extracted from organisms (e.g., wild-type or engineered) that synthesize C50 carotenoids (e.g., microorganisms that synthesize C50 carotenoid compounds). For example, Milon et al., Helv. Chim. Acta 69, 12-24 (1986);Karrer et al., Helv. Chim. Acta 34, 28-33 (1951);Furubayashi et al., Nat Commun. 2015; 6: 7534; Tobias and Arnold, Biochim Biophys Acta. 2006 Feb;1761(2):235-46;Henke et al., (June 14th 2017). Carotenoid Production by Corynebacterium: The Workhorse of Industrial Amino Acid Production as Host for Production of a Broad Spectrum of C40 and C50 Carotenoids, Dragan J. Cvetkovic and Goran S. Nikolic, Intech Open, DOI: 10.5772 / 67631.Available from: intechopen.See, for example, com / books / carotenoids / carotenoid-production-by-corynebacterium-the-workhorse-of-industrial-amino-acid-production-as-host-f; Henke et al., (2017). C50 Carotenoids: Occurrence, Biosynthesis, Glycosylation, and Metabolic Engineering for their Overproduction. In, Bio-pigmentation and Biotechnological Implementations, pp.107-126. doi:10.1002 / 9781119166191.ch5; Fernandes, Introductory Chapter: Carotenoids - A Brief Overview on Its Structure, Biosynthesis, Synthesis, and Applications. In Progress in Carotenoid Research. (2018) doi:10.5772 / intechopen.79542; Heider et al., Appl Microbiol Biot 2014;98(10):4355e68; Wang et al., Biotechnol Adv 2007;25(3):211e22; Niu et al., Synthetic and Systems Biotechnology 2 (2017) 167e175; Li et al., Scientific Reports volume 9, Article number: 2982 (2019); US20050260699; US20140170700; US20040091958; US20090197321; etc.

[0112] In some embodiments, one or more carotenoid compounds for use in accordance with the present disclosure can be provided as purified or substantially purified molecules, or as less purified (e.g., enriched) extracts of carotenoid-producing organisms.

[0113] In some embodiments, preparations that are or include one or more carotenoid compounds are incorporated into or used to produce pharmaceutical compositions described herein that, when administered to a subject, deliver the carotenoid compounds to the subject.

[0114] In some embodiments, a carotenoid preparation or carotenoid composition (e.g., a pharmaceutical composition that contains or delivers a carotenoid compound) comprises at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, or 99% w / w of a carotenoid compound.

[0115] In some embodiments, the composition for use according to the present disclosure is, for example, a pharmaceutical composition for oral administration.The pharmaceutical composition typically comprises an active agent (for example, a carotenoid compound, for example, a C50 carotenoid compound, or a source thereof) and a pharmaceutically acceptable carrier.Some exemplary pharmaceutically acceptable carriers include, for example, physiological saline, solvent, dispersion medium, coating agent, antibacterial and antifungal agent, isotonic and absorption delaying agent, etc., that are compatible with pharmaceutical administration.

[0116] In some embodiments, pharmaceutical compositions for use in accordance with the present disclosure can include and / or be administered with one or more additional active compounds; in certain embodiments, such additional active agents include ginger, curcumin, probiotics (e.g., one or more probiotic strains of the following genera: Lactobacillus, Bifidobacterium, Saccharomyces, Enterococcus, Streptococcus, Pediococcus, Leuconostoc, Bacillus, and / or Escherichia coli (Fijan, Int J Environ Res Public Health. 2014 May; 11(5): 4745-4767); prebiotics (non-digestible food ingredients that help support the growth of probiotic bacteria, such as fructans, e.g., fructooligosaccharides (FOS) and inulin, galactans, e.g., galactooligosaccharides (GOS), dietary fibers, e.g., resistant starch, pectin, beta-glucan, and xylooligosaccharides (Hutkins et al., Curr Opin Biotechnol. 2016 Feb;37:1-7)), and combinations thereof.

[0117] Pharmaceutical compositions are typically formulated to be compatible with their intended route of administration, an example of which is oral administration.

[0118] Methods for formulating suitable pharmaceutical compositions are known in the art, see, for example, Remington: The Science and Practice of Pharmacy, 21st ed., 2005; and Drugs and the Pharmaceutical Sciences: a Series of Textbooks and Monographs (Dekker, NY) series of books. Oral compositions generally include an inert diluent or an edible carrier. In some embodiments, oral formulations can be or include syrup, liquid, tablet, lozenge, gummy, capsule (e.g., gelatin capsule), powder, gel, film, etc., to name just a few examples.

[0119] In some embodiments, pharmaceutically compatible binders and / or auxiliary substances can be included as part of pharmaceutical compositions.In some specific embodiments, pharmaceutical compositions can include, for example, any one or more of the following inactive ingredients or compounds of similar nature: binders, such as microcrystalline cellulose, tragacanth gum, or gelatin; excipients, such as starch or lactose, disintegrants, such as alginic acid, Primogel, or cornstarch; lubricants, such as magnesium stearate or sterotes; glidants, such as colloidal silicon dioxide; sweeteners, such as sucrose or saccharin; or flavorings, such as peppermint, methyl salicylate, or orange flavoring.In some embodiments, the composition can be ingested directly, or can be sprinkled on food or liquid (for example, water), or can be mixed into food or liquid (for example, water).

[0120] In some embodiments, the carotenoid compositions described herein that can be administered to a subject can be or include a ingestible item (e.g., a food or beverage) that contains one or more carotenoids (e.g., is supplemented with one or more carotenoids). In some embodiments, useful compositions can include one or more carotenoid compounds at levels higher than those found in its components and / or higher than those normally present in the associated food or beverage, e.g., to provide carotenoid levels that are sufficient to provide the therapeutic effects described herein.

[0121] In some embodiments, the food can be or include one or more of bars, candy, baked goods, cereals, savory snacks, pasta, chocolate, and other solid foods, as well as liquid or semi-solid foods such as yogurt, soups and stews, and beverages such as smoothies, shakes, juices, and other carbonated or non-carbonated beverages. In some embodiments, the food is provided with the carotenoid already contained therein; in some embodiments, the food is prepared by the subject by mixing in the carotenoid.

[0122] The compositions can be included in a kit, container, pack, or dispenser together with instructions for administration or for use in the methods described herein.

[0123] Those skilled in the art will recognize, upon reading this disclosure, that in some embodiments, the carotenoid compositions described herein can be or can include one or more cells, tissues, or organisms (e.g., plant or microbial cells, tissues, or organisms) that produce (e.g., have produced and / or are producing) the associated compound. In some embodiments, such cells, tissues, or organisms may have previously produced the associated carotenoid; in some embodiments, such cells, tissues, or organisms are producing a carotenoid compound.

[0124] In some embodiments, the carotenoid composition can comprise killed (e.g., heat-killed) cells, tissues, and / or organisms, or in some embodiments, the carotenoid composition can comprise viable or living cells, tissues, and / or organisms.

[0125] In some embodiments, the methods of treatment described herein include administering one or more viable or living cells, tissues, or organisms that synthesize carotenoid compounds. In some such embodiments, the cells, tissues, or organisms are microbial cells and are administered according to a regimen that achieves population of the subject's microbiome with the administered cells.

[0126] In some embodiments, the carotenoid compositions described herein comprise and / or are formulated through the use of one or more cell cultures and / or supernatants or pellets thereof, and / or powders formed therefrom.

[0127] Those skilled in the art will recognize that in some embodiments, techniques for preparing carotenoid compositions and / or preparations and / or for preparing one or more carotenoid compositions (particularly for preparing pharmaceutical compositions) can include one or more steps of evaluating or characterizing the compound, preparation, or composition, e.g., as part of quality control. In some embodiments, if the assayed material does not meet predetermined specifications for the relevant evaluation, it is discarded. In some embodiments, if such assayed material meets predetermined specifications, it continues to be processed as described herein.

[0128] How to identify and / or characterize Among other things, the present disclosure provides a system that allows the evaluation of the properties of one or more agents that are related to the usefulness described herein.In some embodiments, the technique for identifying and / or characterizing the agent described herein can include comparison with appropriate reference (for example, with positive control reference and / or with negative control reference).In some embodiments, reference can be or include historical reference; in some embodiments, reference can be or include contemporaneous reference.

[0129] In some embodiments, the provided technology may be useful for screening test agents, which may be or may include, for example, one or more polypeptides, peptides, inorganic or organic macromolecules or small molecules, or compositions that contain or deliver them, to identify agents useful in the methods described herein. Alternatively, or in addition, in some embodiments, the provided technology may be useful for characterizing one or more agents, for example, during the development and / or commercialization of such agents or pharmaceutically acceptable compositions thereof.

[0130] As used herein, "small molecule" refers to a small organic or inorganic molecule having a molecular weight below about 3,000 Daltons. Generally, a small molecule can have a molecular weight less than 3,000 Daltons (Da). Small molecules can be, for example, at least about 100 Da to about 3,000 Da (e.g., between about 100 to about 3,000 Da, about 100 to about 2,500 Da, about 100 to about 2,000 Da, about 100 to about 1,750 Da, about 100 to about 1,500 Da, about 100 to about 1,250 Da, about 100 to about 1,000 Da, about 100 to about 750 Da, about 100 to about 500 Da, about 200 to about 1,500, about 500 to about 1,000, about 300 to about 1,000 Da, or about 100 to about 250 Da).

[0131] Those skilled in the art will recognize that in some embodiments, particularly in screening embodiments, the provided techniques can be utilized to identify (e.g., screen) and / or characterize a plurality of agents. In some embodiments, such a plurality is or includes reasonably equivalent agents (e.g., one or more specific small molecule compounds and multiple analogs thereof); in some embodiments, the plurality of agents is or includes a plurality of natural products (e.g., carotenoid compounds, such as C50 carotenoid compounds) and / or one or more analogs thereof. In some embodiments, the plurality of agents is or includes a combinatorial library of small molecule compounds. Suitable combinatorial techniques for synthesizing small molecules are known in the art, and include techniques such as "split-pool" or "parallel" synthesis, solid-phase and solution-phase techniques, and encoding techniques, as exemplified, for example, by Obrecht and Villalgordo, "Solid-Supported Combinatorial and Parallel Synthesis of Small-Molecular-Weight Compound Libraries," Pergamon-Elsevier Science Limited (1998) (see, for example, Czarnik, Curr. Opin. Chem. Bio. 1:60-6 (1997)). Furthermore, some small molecule libraries are commercially available. Some suitable small molecule test compounds are listed in U.S. Patent No. 6,503,713, the entire contents of which are incorporated herein by reference.

[0132] In some embodiments, the provided techniques can be used to screen and / or evaluate multiple agents covering a range of features such as charge, aromaticity, hydrogen bonding, flexibility, size, side chain length, hydrophobicity, and rigidity.

[0133] In some embodiments, the libraries screened as described herein can include various types of test compounds. In some embodiments, a given library can include a set of structurally related or unrelated test compounds. In some specific embodiments, the library can include a set of peptide or peptidomimetic molecules. In some embodiments, the library can include carotenoid compounds, for example, naturally occurring or synthetic carotenoids, such as C35, C40, C45, C50, C55, or C60 carotenoids.

[0134] In some embodiments, the provided techniques are utilized to evaluate a set of agents that are related to each other by systematic variation of the structure of the first agent; in some such instances, the first agent can be or can include a compound of known activity (e.g., a carotenoid compound, such as a C50 carotenoid compound).

[0135] In some embodiments, the technology provided herein utilizes or provides a correlation between structural features and the presence (or level) of a biological activity of interest—i.e., a structure-function relationship. In some cases, the structure-function relationship can be defined empirically; in some embodiments, the structure-function relationship can be defined through the use of computer modeling and / or analytical prediction methodologies.

[0136] In some embodiments, the food aversion assay described herein is used. In some embodiments, the test sample is an in vivo model of the disorder described herein or derived from the model (e.g., a sample obtained from the model). For example, a test compound is applied to a test sample containing one or more C. elegans organisms, and one or more effects of the test compound are evaluated. For example, the ability of the test compound to attenuate food aversion in the presence of a chemotherapeutic agent or other nausea-inducing stimulus can be tested. Alternatively, the effect of a reporter gene, such as the ABC transporter gene fusion pgp-5p::gfp, can be evaluated by detecting changes in GFP fluorescence. As those skilled in the art will recognize, other reporters can be easily used.

[0137] In some embodiments, compounds can be screened by the methods described herein to determine whether they can reduce nausea (e.g., have anti-nausea activity), emesis (e.g., have anti-emetic activity), and / or food aversion in a system (e.g., an animal model, e.g., C. elegans). In some embodiments, a compound determined to reduce nausea (e.g., have anti-nausea activity), emesis (e.g., have anti-emetic activity), and / or food aversion in a system (e.g., an animal model, e.g., C. elegans) can be considered a candidate compound. For example, a candidate compound screened in a system having characteristics associated with nausea, vomiting, and / or food aversion, e.g., an in vivo model of a disease, disorder, or condition associated with nausea and / or vomiting, e.g., C. elegans, and determined to have a desirable effect on nausea, vomiting, and / or food aversion can be considered a candidate therapeutic agent. In some embodiments, the candidate therapeutic agent can be tested in a larger animal model or in a clinical setting. Once screened in a clinical setting, the candidate therapeutic agent can be a therapeutic agent. Candidate compounds, candidate therapeutic agents, and therapeutic agents can be optionally optimized and / or derivatized and formulated with physiologically acceptable excipients to form pharmaceutical compositions.

[0138] Compounds can be evaluated by the methods described herein to determine whether they can reduce nausea (e.g., have anti-nausea activity), emesis (e.g., have anti-emetic activity), and / or food aversion in a system (e.g., an animal model, e.g., C. elegans). In some embodiments, the present disclosure provides methods for evaluating compounds (e.g., carotenoids) to determine their ability to reduce nausea (e.g., anti-nausea activity), emesis (e.g., anti-emetic activity), and / or food aversion in a system (e.g., an animal model, e.g., C. elegans). In some embodiments, methods for evaluating compounds to determine their ability to reduce nausea (e.g., anti-nausea activity), emesis (e.g., anti-emetic activity), and / or food aversion in a system (e.g., an animal model, e.g., C. elegans) are part of assays (e.g., release tests, stability tests, efficacy tests, etc.) performed for approval or maintenance of approval from a regulatory agency (e.g., U.S. Food and Drug Administration, European Medicines Agency, etc.). In some embodiments, a method of evaluating a compound (e.g., a carotenoid) to determine its ability to reduce nausea (e.g., anti-nausea activity), vomiting (e.g., anti-emetic activity), and / or food aversion in a system (e.g., an animal model, e.g., C. elegans) is part of a manufacturing method. In some embodiments, the evaluation can be performed as part of a screening method. In some embodiments, the system can be an animal system. In some embodiments, the animal system is a model system, e.g., C. elegans, cat, dog, ape, or pig.

[0139] Evaluation compounds determined to reduce nausea (e.g., have anti-nausea activity), vomiting (e.g., have anti-emetic activity), and / or food aversion in a system (e.g., an animal model, e.g., C. elegans) can be systematically varied, e.g., using rational design, to optimize binding affinity, binding activity, specificity, or another parameter. Methods described herein can also be used to screen and / or evaluate such optimization. In some embodiments, the method includes one or more steps known in the art and / or described herein, e.g., identifying one or more hits in a library, subjecting the hits to systematic structural modification to generate a second library of compounds structurally related to the hits, screening the second library using methods described herein, or a combination thereof, to screen a first library of compounds. Thus, in one embodiment, the method comprises screening a first library of compounds using one or more steps known in the art and / or described herein, e.g., identifying one or more hits in the library, subjecting the hits to systematic structural modifications to generate a second library of compounds structurally related to the hits, screening the second library using a method described herein, or a combination thereof.

[0140] An evaluation compound determined to reduce nausea (e.g., have anti-nausea activity), emesis (e.g., have anti-emetic activity), and / or food aversion in a system (e.g., an animal model, e.g., C. elegans) can be considered a candidate therapeutic compound useful for reducing nausea, emesis, and / or food aversion as described herein. Various techniques useful for determining the structure of compounds, such as NMR, mass spectrometry, gas chromatography with an electron capture detector, fluorescence, and absorption spectroscopy, can be used in the methods described herein. The present disclosure provides insights into how compounds determined (e.g., by the methods described herein) to reduce nausea (e.g., have anti-nausea activity), emesis (e.g., have anti-emetic activity), and / or food aversion in a system can be used in methods for treating, preventing, or delaying the onset or progression of the diseases, disorders, and / or conditions described herein. The present disclosure provides insight that compounds determined in a system (e.g., by the methods described herein) to reduce nausea (e.g., have anti-nausea activity), vomiting (e.g., have anti-emetic activity), and / or food aversion can be used in the methods of treating nausea, vomiting, and / or food aversion described herein.

[0141] An evaluation compound determined to reduce nausea (e.g., have anti-nausea activity), vomiting (e.g., have anti-emetic activity), and / or food aversion in a system (e.g., C. elegans) can be evaluated in a second system (e.g., a larger animal, e.g., ape, monkey, cat, dog, pig, etc.). The animal can be monitored for changes due to the presence of the compound, e.g., reduction in nausea or food aversion. In some embodiments, the system is a human, e.g., a human with CINV or RINV, and the parameter is reduction in the severity, frequency, or duration of nausea, vomiting, and / or food aversion. [Example]

[0142] Embodiments provided herein are illustrated in the following examples, which do not limit the scope of the disclosure or the claims.

[0143] material and method The following materials and methods were used in the examples below. system N2 Bristol was the wild-type strain used. The following strains and mutant alleles were used: TIFF2026012220000002.tif56135

[0144] Growth and handling of microorganisms used: 16S ribosomal sequences were amplified using specific primers and sequenced to identify the microorganisms. LB medium and plates were used to culture Kocuria rhizophila, Arthrobacter alilactensis, and Corynebacterium glutamicum and their mutants. 500 ml of overnight culture was plated onto SK medium plates and incubated at room temperature for 2 days before starting the experiment. For experiments involving Kocuria rhizophila wild-type and mutants, Arthrobacter alilactensis, and Corynebacterium glutamicum wild-type and mutants, synchronized L1-larval animals were grown on E. coli OP50-seeded plates until the L4-larval stage or day 1 of adulthood, washed at least five times in M9 buffer, and then transferred to the appropriate bacterial diet.

[0145] Drug Treatment: Hygromycin diluted in M9 to the desired concentration was added to NGM plates containing E. coli OP50 bacteria. Emetine or cisplatin stock solutions were diluted in M9 and added to the desired concentrations to NGM plates containing E. coli OP50 bacteria. 750 μg / ml of K. rhizophila extract was added to NGM plates containing E. coli OP50 bacteria containing the appropriate concentrations of hygromycin, cisplatin, or emetine. For xenobiotic experiments, synchronized L1-stage animals were dropped onto drug-containing plates and scored after 4 days.

[0146] RNAi assay: For RNAi assays, synchronized L1 larval animals of the appropriate genotype were fed the appropriate RNAi clone until they reached adulthood day 1. RNAi-treated animals were then washed at least five times in M9 to remove E. coli bacteria and transferred to K. rhizophila- or E. coli OP50-seeded plates.

[0147] Microscopic observation C. elegans were mounted on agar pads and images were acquired using a Zeiss AXIO Imager Z1 microscope equipped with a Zeiss AxioCam HRm camera and Axiovision 4.6 (Zeiss) software. All fluorescent images shown within the same figure panel were collected together using the same exposure time. Images were converted to 8-bit images, thresholded, and quantified using ImageJ. Statistical significance was determined using a Student's t-test. Low-magnification bright-field and GFP fluorescent images were acquired using a Zeiss AxioZoom V16 equipped with a Hamamatsu Orca flash 4.0 digital camera and Axiovision ZEN software.

[0148] Multiple alignment of protein sequences Multiple alignments were performed using Clustal Omega software.

[0149] EMS mutagenesis screening of K. rhizophila Mutagenesis was performed by treating overnight cultures of K. rhizophila with 50 mM EMS in PBS at 37°C for 45 minutes. Serial dilutions of the mutagenized K. rhizophila culture were plated on LB medium plates, and approximately 2,000 mutagenized bacterial colonies were selected and grown in LB medium. 500 ml of the overnight culture was inoculated onto SK medium plates and incubated at room temperature for 2 days before the start of the experiment. Synchronized L1-larval stage eft-3(q145);pgp-5p::gfp animals were grown in E. coli OP50-seeded plates until the L4-larval stage or day 1 of adulthood, washed at least five times in M9 buffer, and then transferred to the mutant K. rhizophila bacterial diet. After 24 hours, plates were visually screened for GFP induction.

[0150] Identification of EMS-induced mutations by whole-genome sequencing Genomic DNA extraction, library preparation, Illumina MiniSeq sequencing, and bioinformatics were all performed by The Sequencing Center in Fort Collins, Colorado.

[0151] Isolation of carotenoids from K. rhizophila Carotenoids from K. rhizophila were isolated as described (Giuffrida et al., 2016) with the following modifications. K. rhizophila cultures grown in LB solution were centrifuged at 4000 RPM for 15 minutes and washed with an equal volume of water. After centrifugation to remove the water, an equal volume of acetone was added and centrifuged again at 4000 RPM for 15 minutes. After acetone removal, the sample was wrapped in aluminum foil to protect it from light, and the bacterial pellet was extracted with methanol in a water bath at 65°C. The sample was extracted multiple times with methanol until all cells were bleached. The supernatant was filtered through Whatman No. 1 filter paper. Two volumes of 15% sodium chloride were added to the methanol extract, mixed, and then an equal volume of hexane was added. Yellow carotenoids separated from the methanol-salt mixture and accumulated in the hexane fraction. The hexane fraction was removed and washed at least three times with water. The hexane fraction was evaporated, and the resulting carotenoid pellet was dissolved in methanol.

[0152] High-performance liquid chromatography The crude methanol extract was separated on a 5-micron particle size Agilent Eclipse Plus C18 4.6 x 250 mm column using an Agilent 1200 HPLC equipped with a diode array detector, autosampler, column oven, solvent degasser, and binary pump. The mobile phase was (A) water vs. (B) methanol at a flow rate of 2 ml / min. The column was pre-equilibrated with 90% B at 40°C before sample injection. Following injection, the column was washed isocratically with 90% B for 5 min, then gradient to 100% B over 5 min. The absorbance spectrum of the eluate was monitored from 300 to 700 nm.

[0153] Pull-down experiments using rat liver To identify carotenoid-binding proteins, we used the protocol of Pilbrow et al., 2014, with the following modifications. Protein extracts were obtained by mincing and homogenizing approximately 10 grams of adult rat liver using Roche Tissue-in T-PER tissue protein extraction reagent containing protease inhibitors. The extracts were defatted using methanol-chloroform. Approximately 10 mg of carotenoids from K. rhizophila were incubated with 1 gram of protein extract at 22°C for 1 hour. Unbound carotenoids were removed by size-exclusion chromatography using a Bio-spin P-6 (6K MWCO). The carotenoid-binding protein extract was applied to a DEAE anion-exchange Sepharose resin column pre-equilibrated with anion-exchange buffer A (0.05 M dibasic sodium phosphate, pH 8.0) at 4°C. The sample was allowed to flow through the column by gravity, and the column was washed with anion-exchange buffer A. The protein was eluted with 0.5 ml of 0.5 M NaCl. The yellow fractions were pooled, dialyzed to remove salt, and concentrated using a Vivaspin centrifugal concentrator column. The yellow fractions were separated by 4-12% Native PAGE, and the visible yellow-orange band was excised and subjected to mass spectrometry analysis for protein identification.

[0154] A putative "decaprenoxanthin" carotenoid biosynthetic cluster from microorganisms The putative carotenoid biosynthetic clusters of the following species are highly similar to those of the K. rhizophila genome, have similar sizes, and show the same organization as those of the K. rhizophila genome: Leifsonia xyli (Lxx15630, Lxx15620, Lxx15610, Lxx15600, Lxx15590, and Lxx15580) (Monteiro-Vitorello et al., 2004), Microbacterium testaceum (MTES_3133, MTES_3132, MTES_3131, MTES_3130, MTES_3129, and MTES_3128) (Morohoshi et al., 2011), Cellvibrio gilvus (Morohoshi et al., 2011), and Bacillus subtilis (Morohoshi et al., 2011). gilvus (Celgi_1516, Celgi_1515, Celgi_1514, Celgi_1513, Celgi_1512, and Celgi_1511) (Christopherson et al., 2013), Cellulomonas fimi (Celf_3171, Celf_3170, Celf_3169, Celf_3168, Celf_3167, and Celf_3166) (Christopherson et al., 2013), Sanguibacter kediyeii (Sanguibacter keddieii (Sked_12750, Sked_12760, Sked_12770, Sked_12780, Sked_12790, and Sked_12800) (Ivanova et al., 2009), Jonesia denitrificans (Jden_0342, Jden_0341, Jden_0340, Jden_0339, Jden_0338, and Jden_0337) (Pukall et al., 2009), Mycetocola manganoxydans (Mycetocolamanganoxydans (D9V29_RS08865, D9V29_RS08870, D9V29_RS08875, D9V29_RS08880, D9V29_RS08885, and D9V29_RS08890), Mycetocola miduiensis (BM197_RS02470, BM197_RS02475, BM197_RS02480, BM197_RS02485, BM197_RS02490, and BM197_RS02495), Cryobacterium psychrotolerans ( psychrotolerans (BLQ39_RS02180, BLQ39_RS02185, BLQ39_RS02190, BLQ39_RS02195, BLQ39_RS02200, and BLQ39_RS02205), Subtercola boreus (B7R21_RS02695, B7R21_RS02700, B7R21_RS02705, B7R21_RS02710, B7R21_RS02715, and B7R21_RS02720), Herbiconiux solani (B7R21_RS02695, B7R21_RS02700, B7R21_RS02705, B7R21_RS02710, B7R21_RS02715, and B7R21_RS02720), and solani (HSO01S_RS07000, HSO01S_RS07005, HSO01S_RS07010, HSO01S_RS07015, HSO01S_RS07020, and HSO01S_RS07025), Microbacterium phyllosphaerae (D3H67_RS09120, D3H67_RS09125, D3H67_RS09130, D3H67_RS09135, D3H67_RS09140, and D3H67_RS09145), Leifsonia aquatica (D3H67_RS09146, D3H67_RS09147, D3H67_RS09148, and D3H67_RS09149), and aquatica) (N136_RS22055, N136_RS22060, N136_RS22065, N136_RS22070, N136_RS22075, and N136_RS22080), Microbacterium esteraromaticum (Microbacteriumesteraromaticum) (B4U78_RS09520, B4U78_RS09525, B4U78_RS09530, B4U78_RS09535, B4U78_RS09540, and B4U78_RS09545), Plantibacter species H53 (A4X17_RS18565, A4X17_RS18570, A4X17_RS18575, A4X17_RS18580, A4X17_RS18585, and A4X17_RS18590), Curtobacterium species (ASF23_RS14315, ASF23_RS14320, ASF23_RS14325, ASF23_RS14330, ASF23_RS14335, and ASF23_RS14340), Microterricola pindariensis, pindariensis (GY24_RS04745, GY24_RS04750, GY24_RS04755, GY24_RS04760, GY24_RS04765, and GY24_RS04770), Frondihabitans species (EDF46_RS08000, EDF46_RS08005, EDF46_RS08010, EDF46_RS08015, EDF46_RS08020, and EDF46_RS08025), Salinibacterium scinjungens ( xinjiangense (SAMN06296378_0676, SAMN06296378_0677, SAMN06296378_0678, SAMN06296378_0679, SAMN06296378_0680, and SAMN06296378_0681), Agromyces species (AVP42_RS01110, AVP42_RS01115, AVP42_RS01120, AVP42_RS01125, AVP42_RS01130, and AVP42_RS01135), Microbacterium burkholderia (Microbacterium barkeri) (MBR4_RS00310, MBR4_RS00315, MBR4_RS00320, MBR4_RS00325, MBR4_RS00330, and MBR4_RS00335), Arthrobacter korensis (Arthrobacterkoreensis (BN2404_RS04370, BN2404_RS04375, BN2404_RS04380, BN2404_RS04385, BN2404_RS04390, and BN2404_RS04395), Cryobacterium roopkundense (GY21_RS00565, GY21_RS00570, GY21_RS00575, GY21_RS00580, GY21_RS00585, and GY21_RS00590), Microbacterium oxydans (Microbacterium oxydans (RN51_RS07325, RN51_RS07330, RN51_RS07335, RN51_RS07340, RN51_RS07345, and RN51_RS07350), Arthrobacter luteolus (AL3_RS02570, AL3_RS02575, AL3_RS02580, AL3_RS02585, AL3_RS02590, and AL3_RS02595), Cryobacterium aureum (AL3_RS02590, AL3_RS02595), and aureum (CJ028_RS03575, CJ028_RS03580, CJ028_RS03585, CJ028_RS03590, CJ028_RS03595, and CJ028_RS03600), Curtobacterium ammoniigenes (CAM01S_RS13455, CAM01S_RS13460, CAM01S_RS13465, CAM01S_RS13470, CAM01S_RS13475, and CAM01S_RS13480), Oerskovia enterophila (Oerskovia enterophila) (OJAG_RS08920, OJAG_RS08925, OJAG_RS08930, OJAG_RS08935, OJAG_RS08940, and OJAG_RS08945), Microbacterium paraoxydans (SAMN04489809_1122, SAMN04489809_1123, SAMN04489809_1124, SAMN04489809_1125, SAMN04489809_1126, and SAMN04489809_1127), Agromyces subbeticus (H521_RS21795, H521_RS21800, H521_RS0106365, H521_RS0106370, H521_RS21805, and H521_RS0106380), Arthrobacter crystallopoietes (H521_RS21795, H521_RS21800, H521_RS0106365, H521_RS0106370, H521_RS21805, and H521_RS0106380), and Arthrobacter crystallopoietes (D477_RS18370, D477_RS18375, D477_RS18380, D477_RS18385, and D477_RS18390, D477_RS18395), Georgiannia satyanarayanai (DSZ44_RS04745, DSZ44_RS04750, DSZ44_RS04755, DSZ44_RS04760, DSZ44_RS04765, and DSZ44_RS04770), Microbacterium trichothecinollyticum (Microbacterium trichothecenolyticum (RS82_RS03115, RS82_RS03120, RS82_RS03125, RS82_RS03130, RS82_RS03135, and RS82_RS03140), Arthrobacter woluwensis (C6401_RS03950, C6401_RS03955, C6401_RS03960, C6401_RS03965, C6401_RS03970, and C6401_RS03975), Promicromonospora kloppenstettii (Promicromonospora kroppenstedtii) (PROKR_RS13815, PROKR_RS13820, PROKR_RS13825, PROKR_RS13830, PROKR_RS13835, and PROKR_RS13840), Cellulomonas ceracea (Cellulomonascellasea (Q760_RS04595, Q760_RS04600, Q760_RS04605, Q760_RS18340, Q760_RS04615, and Q760_RS04620), Agromyces cerinus (BUR99_RS12060, BUR99_RS12065, BUR99_RS12070, BUR99_RS12075, BUR99_RS12080, and BUR99_RS12085), Agreia platensis (Agreia pratensis (B9Y86_RS06900, B9Y86_RS06905, B9Y86_RS06910, B9Y86_RS06915, B9Y86_RS06920, and B9Y86_RS06925), Microbacterium laevaniformans (OR221_3062, OR221_3063, OR221_3064, OR221_3065, OR221_3066, and OR221_3067), Arthrobacter stuckebrantii (Arthrobacter stackebrandtii) (CVV67_17780, CVV67_17785, CVV67_17790, CVV67_17795, CVV67_17800, and CVV67_17805), Paeniglutamicibacter gangotriensis (Paeniglutamicibacter gangotriensis (ADIAG_RS03760, ADIAG_RS03765, ADIAG_RS03770, ADIAG_RS03775, ADIAG_RS03780, and ADIAG_RS03785), Microbacterium trichothecinolyticum (RS82_RS03115, RS82_RS03120, RS82_RS03125, RS82_RS03130, RS82_RS03135, and RS82_RS03140), Arthrobacter livingstonensis (Arthrobacter livingstonensis) (CVV68_RS19330, CVV68_RS19335, CVV68_RS19340, CVV68_RS19345, CVV68_RS19350, and CVV68_RS19355), Demequina lutea (lutea (AOP76_RS09030, AOP76_RS09035, AOP76_RS09040, AOP76_RS09045, AOP76_RS09050, and AOP76_RS09055), Zhihengliuella halotolerans (CUR88_RS12685, CUR88_RS12690, CUR88_RS12695, CUR88_RS12700, CUR88_RS12705, and CUR88_RS12710), Paeniglutamicibacter antarctica antarcticus (BN2261_RS08280, BN2261_RS08285, BN2261_RS08290, BN2261_RS08295, BN2261_RS08300, and BN2261_RS08305), Janibacter melonis (EEW87_RS00715, EEW87_RS00720, EEW87_RS00725, EEW87_RS00730, EEW87_RS00735, and EEW87_RS00740), Microbacterium arborescens ( arborescens) (DOU46_RS02280, DOU46_RS02285, DOU46_RS02290, DOU46_RS02295, DOU46_RS02300, and DOU46_RS02305), Aglaia platensis (B9Y86_RS06900, B9Y86_RS06905, B9Y86_RS06910, B9Y86_RS06915, B9Y86_RS06920, and B9Y86_RS06925), Aglaia bicolorata (Agreia bicolorata) (TZ00_RS04480, TZ00_RS04485, TZ00_RS19215, TZ00_RS04495, TZ00_RS04500, and TZ00_RS04505), Arthrobacter cyclotectophilus (Arthrobacterpsychrochitiniphilus (CVS30_RS02785, CVS30_RS02790, CVS30_RS02795, CVS30_RS02800, CVS30_RS02805, and CVS30_RS02810), Microtelicola pindariensis (GY24_RS04745, GY24_RS04750, GY24_RS04755, GY24_RS04760, GY24_RS04765, and GY24_RS04770), Microbacterium indicum (Microbacterium indicum (H576_RS15860, H576_RS0112930, H576_RS0112935, H576_RS0112940, H576_RS0112945, and H576_RS15865), Homoserinimonas species (DL891_RS01870, DL891_RS01875, DL891_RS01880, DL891_RS01885, DL891_RS01890, and DL891_RS01895), Cryobacterium levicorallinum (Cryobacterium levicorallinum (SAMN05216274_11068, SAMN05216274_11069, SAMN05216274_11070, SAMN05216274_11071, SAMN05216274_11072, and SAMN05216274_11073), Frigoribacterium species (EDF18_RS14355, EDF18_RS14360, crtI, EDF18_RS14370, EDF18_RS14375, and EDF18_RS14380), Cryobacterium luteum (Cryobacterium luteum) (SAMN05216281_10883, SAMN05216281_10884, SAMN05216281_10885, SAMN05216281_10886, SAMN05216281_10887, and SAMN05216281_10888), Cellulomonas carbonis (Cellulomonascarbonis (N868_RS13600, N868_RS13605, N868_RS13610, N868_RS13615, N868_RS13620, and N868_RS13625), Okibacterium fritillariae (B5X75_RS14075, B5X75_RS14080, B5X75_RS14085, B5X75_RS14090, B5X75_RS14095, and B5X75_RS14100), Glycomyces sambucus (Glycomyces sambucus (BLS99_RS13650, BLS99_RS13655, BLS99_RS13660, BLS99_RS13665, BLS99_RS13670, and BLS99_RS13675), Krasilnikoviella flava (B5Y66_RS20515, B5Y66_RS20520, B5Y66_RS20525, B5Y66_RS20530, B5Y66_RS20535, and B5Y66_RS20540), Actinotalea ferrariae (B5Y66_RS20515, B5Y66_RS20520, B5Y66_RS20525, B5Y66_RS20530, B5Y66_RS20535, and B5Y66_RS20540), and Actinotalea ferrariae (N866_01505, N866_01510, N866_01515, N866_01520, N866_01525, and ubiA), Lysinimicrobium soli (AOM04_RS11780, AOM04_RS11785, AOM04_RS11790, AOM04_RS11795, AOM04_RS11800, and AOM04_RS11805), Luteimicrobium subalcuticum (AOM04_RS11806, AOM04_RS11807, AOM04_RS11808, AOM04_RS11809, AOM04_RS11810, AOM04_RS11811, AOM04_RS11812, AOM04_RS11813, AOM04_RS11814, AOM04_RS11815, AOM04_RS11816, AOM04_RS11817, AOM04_RS11818, AOM04_RS11819, AOM04_RS11820, AOM04_RS11821, AOM04_RS11822, AOM04_RS11823, AOM04_RS11824, AOM04_RS11825, AOM04_RS11826, AOM04_RS11827, AOM04_RS11828, AOM04_RS11829, AOM04_RS11930, AOM04_RS11931, AOM04_RS11932, AOM04_RS11933, AOM04_RS11934, AOM0 subarcticum (CLV34_RS08275, CLV34_RS08280, CLV34_RS08285, CLV34_RS08290, CLV34_RS08295, and CLV34_RS08300), Promicromonospora kloppenstettii (PROKR_RS13815, PROKR_RS13820, PROKR_RS13825, PROKR_RS13830, PROKR_RS13835, and PROKR_RS13840), Tersicoccus phoenicis (Tersicoccusphoenicis (BKD30_RS05860, BKD30_RS05865, BKD30_RS05870, BKD30_RS05875, BKD30_RS05880, and BKD30_RS05885), Sinomonas humi (LK10_RS09295, LK10_RS09300, LK10_RS09305, LK10_RS09310, and LK10_RS09315), Pseudarthrobacter phenanthrenivorans ( phenanthrenivorans (RM50_RS01675, RM50_RS01680, RM50_RS01685, RM50_RS01690, RM50_RS01695, and RM50_RS01700), Acaricomes phytoseiuli (C501_RS0107225, C501_RS0107230, C501_RS0107235, C501_RS0107240, C501_RS0107245, and C501_RS0107250), Leucobacter musalum (Leucobacter musarum) (AMS67_RS10795, AMS67_RS10800, AMS67_RS10805, AMS67_RS10810, AMS67_RS10815, and AMS67_RS10820), Ornithinimicrobium pekingens ( pekingense (K330_RS19765, K330_RS0107130, K330_RS19770, K330_RS19775, K330_RS0107145, and K330_RS0107150), Citricoccus species (CITRI_RS16000, CITRI_RS0102550, CITRI_RS0102555, CITRI_RS16005, CITRI_RS0102565, and CITRI_RS0102570), and Arthrobacter aritense (Arthrobacter arilatensis)(AARI_13710, AARI_13720, AARI_13730, AARI_13740, AARI_13760, and AARI_13750)(Monnet et al.,In Corynebacterium glutamicum (cg0723, cg0721, cg0720, cg0719, cg0718, and cg0717) (Kalinowski et al., 2003) and Corynebacterium efficiens (HMPREF0290_1086, HMPREF0290_1088, HMPREF0290_1089, HMPREF0290_1090, HMPREF0290_1091, and HMPREF0290_1092) (Nishio et al., 2003), the decaprenoxanthin-producing gene cluster is located in the crtE and crtB regions of C. glutamicum or the crtB region of C. efficiens. The clusters are similar in size and organization to those of K. rhizophylla, except for the insertion of an unrelated gene, cg0722, between HMPREF0290_1088 in Corynebacterium efficiens. Furthermore, additional carotenoid clusters (NCgl0600, NCgl0598, NCgl0597, NCgl0596, NCgl0595, and NCgl0594) are also present in the genome of Corynebacterium glutamicum. In Kytococcus sedentarius, genes involved in carotenogenesis (Ksed_13840, Ksed_13830, Ksed_13820, Ksed_13810, Ksed_13800) are arranged in the same cluster, whereas Ksed_16070, encoding geranylgeranyl pyrophosphate synthase, is located elsewhere in the genome (Sims et al., 2009).

[0155] In Brevibacterium mcbrellneri, genes involved in carotenoid production (HMPREF0183_0793, HMPREF0183_0794, HMPREF0183_0795, HMPREF0183_0796, and HMPREF0183_0797) are located in the same cluster, whereas HMPREF0183_0437, encoding a polyprenyl synthetase, is located elsewhere in the genome.

[0156] In Beutenbergia cavernae, genes involved in carotenoid production (Bcav_3492, Bcav_3491, Bcav_3490, Bcav_3489, Bcav_3488) are located in the same cluster, whereas Bcav_0970, encoding a polyprenyl synthetase, is located elsewhere in the genome (Land et al., 2009).

[0157] In Brachybacterium faecium (Bfae_04470, Bfae_04440, Bfae_04430, Bfae_04420, Bfae_04410, and Bfae_04400) (Lapidus et al., 2009), the carotenoid biosynthetic cluster is similar in size and organization to that of K. rhizophila, except for the insertion of two unrelated genes, Bfae_04460 and Bfae_04450, between Bfae_04470 and Bfae_04440.

[0158] In Cellulomonas flavigna (Cfla_2888, Cfla_2889, Cfla_2890, Cfla_2891, and Cfla_2892) (Abt et al., 2010), all genes involved in carotenoid production are present, except for Cfla_2893, which may be a pseudogene due to a frameshift mutation.

[0159] Decaprenoxanthin was the first C50 carotenoid discovered in Flavobacterium dehydrogenans (now known as Agromyces mediolanus (Liaaen-Jensen et al., 1968)).

[0160] Many bacteria are known to produce decaprenoxanthin, including Agromyces mediolanus (Liaaen-Jensen et al., 1968), Aureobacterium species (Fukuoka et al., 2004), Arthrobacter glacialis (Arpin et al., 1975), Arthrobacter alylatensis (Sutthiwong et al., 2014), Cellulomonas biazotea (Weeks et al., 1980), Citriococcus species, and Corynebacterium glutamicum (Krubasik et al., 2001).

[0161] Example 1. Bacterial carotenoids suppress C. elegans surveillance for translation defects Among the approximately 500 C. elegans xenobiotic detoxification genes, expression of a specific set of cytochrome p450, ABC transporter, and UDP-glycosyltransferase genes, such as the C. elegans ABC transporter gene pgp-5, can be induced by toxins, RNAi, or mutational suppression of ribosomal proteins, tRNA synthetases, and other genes involved in translation (Govindan et al., 2015). Even when translation defects are restricted to the germline, such as in the C. elegans eft-3(q145) mutant, a mutation in the germline isoform of translation elongation factor-1 (in which somatic translation and development are normal, but germline translation is abolished and the germline cannot proliferate), expression of the ABC transporter gene fusion pgp-5p::gfp was strongly induced in the intestine when animals were fed benign E. coli OP50 (Figure 5A). Feeding C. elegans eft-3(q145);pgp-5p::gfp with K. rhizophila, but not E. coli, disrupted the normal induction of pgp-5p::gfp (Figure 5A & B). K. rhizophila is a Gram-positive coccobacillus in the Actinobacteria phylum, a clade rich in drug biosynthesis pathways. K. rhizophila species are found in various environmental niches, including the gut microbiome of C. elegans in natural populations of soil- and orchard-derived nematodes (Felix et al., 2010). K. rhizophila species are normal inhabitants of the skin and mucous membranes of humans and animals but can be associated with human infections.

[0162] To confirm that K. rhizophila can suppress surveillance of various translation defects, we tested the induction of xenobiotic detoxification responses to RNAi of other ribosomal proteins. Synchronized L1-larval stage pgp-5p::gfp animals were fed E. coli expressing either rpl-1 dsRNA or vrs-2 dsRNA, which inhibit the production of C. elegans RPL-1 ribosomal protein and VRS-2 tRNA synthetase. Once these animals reached adulthood, they were transferred to either E. coli OP50-containing plates or K. rhizophila-seeded plates and scored for GFP induction 24 hours later. In animals fed either vrs-2 dsRNA or rpl-1 dsRNA and transferred to E. coli OP50-containing plates, pgp-5p::gfp was induced. In contrast, expression of pgp-5p::gfp was suppressed in animals given either rpl-1 dsRNA or vrs-2 dsRNA and transferred to K. rhizophila plates ( Figure 5C & D ). The suppression of the surveillance pathway by K. rhizophila is specific to translational stress, as K. rhizophila does not suppress the induction of the mitochondrial stress response or the endoplasmic reticulum stress response ( Govindan et al., 2015 ).

[0163] To identify the K. rhizophila pathway responsible for the repression of pgp-5p::gfp induction in a C. elegans strain carrying a gene defect in translation (eft-3(q145)), we performed a forward genetic screen for K. rhizophila mutant strains defective in the repression of pgp-5p::gfp induction. Approximately 2000 individual K. rhizophila strains that grew normally on bacterial LB plates after EMS mutagenesis were identified as eft-3(q145);p These 2,000 individual wells of different K. rhizophila mutant strains were screened for mutant bacterial strains that did not suppress pgp-5p::gfp induction in eft-3(q145);pgp-5p::gfp animals. Six K. rhizophila mutant strains were identified that did not induce pgp-5p::gfp in eft-3(q145);pgp-5p::gfp animals (Figure 5A & E). Visual inspection revealed that all of these mutant strains were defective in colony pigmentation compared to wild-type K. rhizophila (Figure 5F). While wild-type K. rhizophila was yellow, six mutant colonies were red, white, or orange. Using this discoloration phenotype, approximately 500,000 bacterial colonies generated by EMS mutagenesis were visually screened for mutants with the discoloration phenotype. 71 discolored mutants were isolated (Figure 5G). These mutants, along with 25 control non-discolored mutants generated by the same EMS mutagenesis, were tested in eft-3(q145);pgp-5p::gfp animals and scored for GFP induction. While all 71 discoloration mutants failed to suppress pgp-5p::gfp induction in C. elegans eft-3(q145) mutants, the 25 normally pigmented strains did not. suppressed the induction of pgp-5p::gfp (Fig. 6A; Fig. 1A-C).

[0164] Genome sequencing of 23 EMS-mutagenized K. rhizophila mutants that did not suppress pgp-5p::gfp induction in the eft-3(q145) mutant revealed that each harbored a mutation in one of six carotenoid biosynthetic cluster genes (Figure 6B; Figures 1C-E). Carotenoids are yellow-to-red pigments produced by the terpenoid biosynthetic pathway. The K. rhizophila genome contains an operon encoding predicted carotenoid (crt) biosynthetic genes (Takarada et al., 2008). These include crtE (KRH_20850; encoding a GGPP synthase), crtB (KRH_20840; encoding a phytoene synthase), crtI (KRH_20830; encoding a phytoene desaturase), crtEB (KRH_20800; encoding a lycopene elongase), and crtYE (KRH_20820; encoding a C 50 ), which encodes carotenoid epsilon cyclase, and crtYf (KRH_20810; C 50 The CrtEb and CrtYe / f cyclases encode carotenoid epsilon cyclase (Figure 1C-E). Based on orthology, the reactions catalyzed by GGPP synthase CrtE, phytoene synthase CrtB, and phytoene desaturase CrtI are predicted to mediate the steps in the production of lycopene (Klassen et al., 2010; Krubasik et al., 2001). CrtEb and CrtYe / f cyclases mediate the conversion of C from lycopene. 50 Catalyzes the biosynthesis of carotenoids. 50 Carotenoids are rare in nature and most of them remain uncharacterized (Krubasik et al., 2001a;Krubasik et al., 2001b;Norgard et al., 1970;Tao et al., 2007;Netzer et al., 2010).

[0165] Although there are several microorganisms that contain CrtEb and CrtYe / f genes (Fig. 7), to date, no genetically and biochemically well-characterized C . 50The only carotenoid is decaprenoxanthin from C. glutamicum (Heider et al., 2012). In C. glutamicum, the enzymes CrtEb, CrtYe, and CrtYf convert lycopene to C. 50 This reaction occurs in two steps: CrtEb converts the acyclic C 50 C to the carotenoid flavuxanthin 40 It catalyzes the elongation of acyclic lycopene (Krubasik et al., 2001a; Krubasik et al., 2001B). The products of CrtYe and CrtYf combine to form C 50 cyclase, and then C 50 It catalyzes the conversion of the carotenoid flavuxanthin to decaprenoxanthin (Krubasik et al., 2001a; Krubasik et al., 2001B). The crtYe and crtYf genes of K. rhizophila are 38% and 34% identical to the crtYe and crtYf genes of C. glutamicum, respectively. Therefore, the yellow pigment produced by K. rhizophila is decaprenoxanthin C. 50 -May belong to the subfamily

[0166] A genetic screen of K. rhizophila yielded multiple mutations in crtI, encoding phytoene desaturase, including six missense mutations (e21, e11, e23, e14, e5, and e13) and two nonsense mutations (e15 and e10) (Figure 1C-E). CrtI catalyzes the conversion of uncolored phytoene to red lycopene. All of these crtI mutants were white colonies (Figure 6B; Figure 1C), similar to the ΔcrtI mutant of C. glutamicum (Heider et al., 2012) (Figure 6C), and therefore likely defective in lycopene synthesis. Six missense mutations were located in highly conserved residues, suggesting they may be important for protein function (Figure 8). e4, e6, and e8 were missense mutations in the crtB gene, encoding phytoene synthase (Figure 1D&E). All of these missense mutations were in highly conserved residues, suggesting that they may be important for protein function (Figure 9). These mutants generated white bacterial colonies (Figure 6B; Figure 1C), as observed in the ΔcrtB mutant of C. glutamicum (Heider et al., 2012) (Figure 6C). Four mutations were obtained in crtEb; two nonsense mutations (e16 and e17) and two missense mutations (e3 and e19) ​​in highly conserved residues (Figure 1D; Figure 10). The mutations in crtEb may be defective in the conversion of lycopene to flavuxanthin (Figure 1E). These mutants formed light red colonies (Figure 6; Figure 1B), likely due to the accumulation of lycopene (but not flavuxanthin), as observed in the ΔcrtEb mutant of C. glutamicum (Heider et al., 2012) (Figure 6C). e17 was a premature stop mutation in crtEb predicted to produce a truncated protein of only 13 amino acids (Figure S10). Mutations in crtYe and crtYf were defective in the last step; homologs of these genes catalyze the synthesis of decaprenoxanthin. These mutants produced pale red to orange colonies (Figure S6B).Interestingly, the ΔcrtY mutant of C. glutamicum accumulated flavuxanthin and also exhibited a light orange to red color ( Heider et al., 2012 ) ( Figure 6C ).

[0167] Because C. glutamicum ATCC13032 is known to produce decaprenoxanthin, we tested whether feeding C. glutamicum ATCC13032 would suppress the induction of pgp-5p::gfp in eft-3(q145);pgp-5p::gfp animals. Feeding eft-3(q145);pgp-5p::gfp animals with wild-type C. glutamicum ATCC13032 suppressed the induction of pgp-5p::gfp (Figure 2A & B). In C. glutamicum ATCC13032, the carotenoid gene cluster CrtE-cg0722-CrtBIYeYfEb mediates decaprenoxanthin biosynthesis (Heider et al., 2012). We tested whether C. glutamicum ATCC13032 deletion mutants of crtY, crtEb, crtI, and crtB, known to be defective in decaprenoxanthin production (Heider et al., 2012), could suppress pgp-5p::gfp induction in eft-3(q145);pgp-5p::gfp animals. Animals fed ΔcrtY, ΔcrtEb, ΔcrtI, and ΔcrtB C. glutamicum showed normal pgp-5p::gfp induction, unlike the same strains grown on wild-type C. glutamicum (Figure 2A & B).

[0168] The pigmented bacterium Arthrobacter aliraitensis is known to produce decaprenoxanthin (Monnet et al., 2010); experiments were conducted to test whether feeding A. aliraitensis would suppress the induction of pgp-5p::gfp in eft-3(q145);pgp-5p::gfp animals. Feeding A. aliraitensis to eft-3(q145);pgp-5p::gfp animals also suppressed pgp-5p::gfp expression (Figure 2C). Thus, C. glutamicum, A. aliraitensis, or K. rhizophylla produce pigmented carotenoids that mediate the suppression of translational surveillance for the induction of ABC transporter detoxification responses.

[0169] Carotenoids, such as decaprenoxanthin, are lipophilic molecules that localize to the cell membrane and can be easily extracted in nonpolar solvents. K. rhizophylla cultures were extracted with such solvents (Figure 11). TLC analysis of the extract revealed the presence of yellow-orange pigments (Figure 2D). HPLC analysis of the methanol extract of K. rhizophylla combined at least six distinct components (Figure 11; Figure 2E). According to their elution times, these peaks were named Peak 1 (5.4 min), Peak 2 (6.3 min), Peak 3 (6.5 min), Peak 4 (7.2 min), Peak 5 (7.8 min), and Peak 6 (8.7 min) (Figure 2E). The absorbance spectrum of the eluted peak revealed absorption maxima at 420, 440, and 470 nm, which were similar to the published absorption spectrum of decaprenoxanthin from Arthrobacter (Giuffrida et al., 2016; Sutthiwong et al., 2014).

[0170] To analyze carotenoid production in different K. rhizophila mutants, carotenoids were extracted from the nonsense mutant alleles crtI(e10), crtEb(e17), crtYe(e22), and crtYf(e18) and the missense mutant crtB(e6). Spectrophotometric analysis of methanol extracts of wild-type K. rhizophila showed an absorption maximum between 415 and 425 nm, whereas the crtEb(e17) extract showed an absorption maximum between 445 and 455 nm (Figure 12). Methanol extracts of the K. rhizophila crtEb(e17) and crtYe(e22) mutants showed similar absorption spectra, whereas extracts of K. rhizophila crtI(e10) and crtB(e6) showed no absorption at all. The methanol extract of crtYf(e18) showed two separate absorption peaks, one at about 400 nm and another smaller peak at about 500 nm.

[0171] We also tested the ability of a crude methanol extract of wild-type K. rhizophila, which contains carotenoids, to suppress GFP induction in eft-3(q145);pgp-5p::gfp animals. Animals fed E. coli supplemented with wild-type K. rhizophila extract showed significantly reduced pgp-5p::gfp expression compared with eft-3(q145);pgp-5p::gfp animals fed E. coli with a control methanol extract (Figure 2F). The extract, a methanol extract of wild-type K. rhizophila, was able to rescue the suppression of the C. elegans surveillance defect of K. rhizophila carotenoid biosynthesis mutants: GFP was not induced when K. rhizophila crtEb(e17), crtB(e6), or crtI(e10) mutants supplemented with K. rhizophila wild-type extract were fed to eft-3(q145);pgp-5p::gfp animals, whereas GFP expression was induced by the eft-3 mutants in C. elegans in animals fed a control extract (Figure 2G).

[0172] One possible explanation for the failure of K. rhizophila carotenoid mutants to suppress pgp-5p::gfp in translationally defective C. elegans mutants could be that these K. rhizophila pigmentation mutants are able to induce pgp-5p::gfp even in a wild-type C. elegans background. To test this possibility, we fed wild-type C. elegans carrying the pgp-5p::gfp fusion gene with K. rhizophila crtEb(e17), crtYe(e22), crtYf(e18), crtB(e6), or crtI(e10) mutants. The results showed that K. rhizophila wild-type and carotenoid mutants failed to induce pgp-5p::gfp (Figure 3A). Another possible interpretation was that feeding K. rhizophila could induce other stress responses in C. elegans that somehow "distract" the animals from monitoring translation. The effects of inducing other GFP-fusion reporters of stress were tested in wild-type and various K. rhizophila mutants. hsp-4p::gfp and hsp-6p::gfp, respectively, induce the endoplasmic reticulum unfolded protein response (UPR). ER ) and mitochondrial unfolded protein response (UPR) mito) (Yoneda et al., 2004; Calfon et al., 2002). clec-60 is a C-type lectin / CUB domain protein induced by the Gram-positive pathogens Staphylococcus aureus and M. nematophilum (O'Rourke et al., 2006). F35E12.5p::GFP is a CUB domain protein induced by Y. pestis, M. nematophilum, and P. aeruginosa (O'Rourke et al., 2006; Troemel et al., 2006; Bolz et al., 2010). C. elegans hsp-4p::gfp, HSp-6p::gfp, F35E12.5p::GFP, and clec-60p::gfp were fed to wild-type and carotenoid mutant K. rhizophila. K. rhizophila did not induce expression of hsp-4p::gfp (Figure 13A). Similarly, K. rhizophila wild-type or carotenoid mutant did not induce expression of hsp-6p::gfp (Figure 13B). K. rhizophila wild-type or carotenoid mutant did not induce F35E12.5p::GFP (Figure 13D). However, K. rhizophila wild-type or carotenoid mutant induced clec-60::GFP, which is induced by Gram-positive bacteria (Figure 13C). Because K. rhizophila is also a Gram-positive bacterium, induction of clec-60 is most likely an immune response to the pathogen.

[0173] To address whether the effect of feeding K. rhizophila on the suppression of pgp-5p::gfp induction was reversible, K. rhizophila -fed eft-3(q145);pgp-5p::gfp animals were transferred to plates of E. coli OP50 after various times. GFP expression was restored within 12 hours of transfer (Figure ​(Figure13E).

[0174] Because C50 carotenoids contain multiple conjugated double bonds, they may possess antioxidant activity (Edge et al., 1997). However, for several reasons, the ROS-quenching properties of carotenoids are unlikely to be involved in the suppression of pgp-5p::gfp induction. First, pgp-5p::gfp is not induced by oxidative stress (Govindan et al., 2015). Second, we tested whether known antioxidants could suppress pgp-5p::gfp induction in a translation-defective C. elegans mutant. Eft-3(q145);pgp-5p::gfp animals grown in E. coli OP50 were treated with either N-acetylcysteine, ascorbic acid, trolox, or resveratrol and screened after 50 h at 20°C. Induction of pgp-5p::gfp was not significantly different in antioxidant-treated animals compared with mock-treated eft-3(q145);pgp-5p::gfp animals (Figure 14A). Third, commercially available carotenoids were tested for their ability to suppress pgp-5p::gfp induction in translation-deficient C. elegans mutants. eft-3(q145);pgp-5p::gfp animals grown in E. coli OP50 were treated with either beta-carotene or astaxanthin and screened after 50 hours at 20°C. Induction of pgp-5p::gfp was not significantly different in animals given these antioxidants compared with mock-treated eft-3(q145);pgp-5p::gfp animals (Figure 14B). Finally, eft-3(q145);pgp-5p::gfp animals were fed E. coli expressing either zeaxanthin, neurosporene, violaxanthin, delta-carotene, or alpha-carotene and screened for GFP induction. Induction of pgp-5p::gfp was not significantly different in animals fed E. coli expressing carotenoids compared to mock-treated eft-3(q145);pgp-5p::gfp animals (Figure 14C). None of these carotenoids are C50 class carotenoids.

[0175] K. rhizophila also suppresses the detoxification response to translation-inhibiting drugs. Hygromycin is a bacterially produced antibiotic (derived from Streptomyces hygroscopicus) that inhibits translation and induces xenobiotic detoxification in C. elegans. Hygromycin at 10 μg / ml induces pgp-5p::gfp expression in animals fed E. coli OP50, whereas K. rhizophila and animals fed 10 μg / ml hygromycin did not induce pgp-5p::gfp (Figure 3A; Figure S4D). However, at higher concentrations of hygromycin, pgp-5p::gfp was induced in both E. coli OP50-fed and K. rhizophila-fed animals (Figure S4D). In contrast, pgp-5p::gfp animals fed K. rhizophila crtI(e10) or crtEb(e17) had no effect on GFP induction in response to hygromycin treatment (Figure 3A). Similar results were obtained using emetine, which blocks protein synthesis by binding to the 40S subunit of the ribosome. 6.25 μg / ml emetine induces pgp-5p::gfp expression in animals fed E. coli OP50 (Figure 15A & B); however, K. rhizophila and animals fed 6.25 μg / ml emetine do not induce pgp-5p::gfp. However, at high concentrations of emetine, pgp-5p::gfp is induced in both E. coli OP50-fed and K. rhizophila-fed animals (Figure 15A). In contrast, feeding pgp-5p::gfp animals with K. rhizophila crtI (e10) or crtEb (e17) did not affect GFP induction in response to emetine treatment (Figure 15B). pgp-5p::gfp was activated in response to genotoxic stress induced by cisplatin, which interferes with DNA replication. 1 mM cisplatin induces pgp-5p::gfp expression in animals fed E. coli OP50, but animals fed K. rhizophila and 1 mM cisplatin do not induce pgp-5p::gfp (Figure 15C). In contrast, feeding pgp-5p::gfp animals with K. rhizophila crtI (e10), crtEb (e17), or crtYe (e22) did not affect GFP induction in response to cisplatin treatment (Figure 15C).

[0176] Because K. rhizophila carotenoids suppress the induction of xenobiotic detoxification genes in C. elegans due to translation defects, we evaluated the ability of K. rhizophila carotenoids to enhance the sensitivity of C. elegans to translation inhibitors. While >80% of wild-type animals fed E. coli OP50 and 10 μg / ml hygromycin reached adulthood in 4 days, <10% of animals fed E. coli OP50, 10 μg / ml hygromycin, and K. rhizophila carotenoid extract reached adulthood in 4 days (Figure 3B; Figure S5D). Carotenoids themselves were not toxic to helminths in the absence of hygromycin (Figure 3B; Figure S5D). Similar results were obtained using emetine: animals treated with K. rhizophila extract were hypersensitive to emetine compared to animals fed the control extract (Figure 3C). Furthermore, animals treated with K. rhizophila extract were hypersensitive to cisplatin compared with animals given the control extract (Figure 3D). The xenobiotic hypersensitivity phenotype may not have been a generalized phenomenon, as the K. rhizophila carotenoid extract did not alter the sensitivity of the animals to antimycin, a mitochondrial poison (Figure S5E).

[0177] Food aversion behavior in C. elegans is induced when animals are exposed to xenobiotics or inactivated essential genes (Melo and Ruvkun, 2012). Exposure of animals to hygromycin (a ribosomal translation inhibitor) or cisplatin (a DNA replication inhibitor) induces a strong food aversion. Approximately 40% of animals exposed to 25 μg / ml hygromycin exhibit food aversion behavior, whereas approximately 15% of animals exposed to 25 μg / ml hygromycin and a carotenoid extract of K. rhizophila exhibit aversion (Figure 3E). Similar results were obtained using cisplatin: approximately 50% of animals exposed to 1 mM cisplatin exhibit aversion behavior, whereas <20% of animals exposed to 1 mM cisplatin and a carotenoid extract of K. rhizophila exhibit food aversion (Figure 3F).

[0178] Example 2. C. elegans Pathway Analysis of K. rhizophila Repression of Translational Surveillance To assess how K. rhizophila carotenoids inhibit the induction of the xenobiotic detoxification response pathway, we performed genetic epistasis analysis using a series of C. elegans mutants that disrupt or activate signaling pathways for translational surveillance at various steps (Govindan et al., 2015). The ZIP-2 / bZIP transcription factor is required for the induction of pgp-5p::gfp expression in response to translational repression, representing the final step in transcriptional induction (Govindan et al., 2015). Overexpression of ZIP-2::mCherry under an intestine-specific promoter was sufficient to induce pgp-5p::gfp expression in wild-type C. elegans, even in the absence of translational repression (Figure 4A). We then tested whether feeding K. rhizophila affected the induction of pgp-5p::gfp in ZIP-2::mCherry-overexpressing strains. Induction of pgp-5p::gfp was similar in animals fed E. coli OP50 and K. rhizophila ( Fig. 4A ), suggesting that K. rhizophila carotenoids disrupt a surveillance pathway component (or a parallel pathway) upstream of the ZIP-2 bZIP transcription factor.

[0179] Induction of xenobiotic detoxification genes in C. elegans by translational repression is dependent on the bile acid signaling pathway (Govindan et al., 2015). C. elegans, which have a genetic defect in bile acid biosynthesis, do not activate pgp-5p::gfp in response to eft-3(q145), RNAi of translational components, or G418 drug repression of translation, yet mammalian bile acids can restore this signal (Govindan et al., 2015). Feeding K. rhizophila inhibits the induction of pgp-5p::gfp in eft-3(q145) animals, but the addition of exogenous mammalian bile acids reactivates GFP expression, even in the presence of wild-type K. rhizophila (Figure 4B & C). Thus, K. rhizophila carotenoids act upstream of this translational surveillance and response pathway in C. elegans, or at a bile acid signaling step.

[0180] To determine the mechanistic pathways by which K. rhizophila carotenoids could modulate the bile acid signaling pathway, we performed a cherry-selective RNAi screen for C. elegans homologs of eukaryotic genes mediating carotenoid binding or transport (Table 1). In this screen, eft-3(q145);pgp-5p::gfp animals were fed E. coli expressing dsRNA against C. elegans homologs of carotenoid binding or transport proteins. Once these animals reached adulthood, they were transferred to K. rhizophila-seeded plates and scored for GFP induction 24 hours later. Expression of pgp-5p::gfp was abrogated in animals fed the dsRNA negative control and then transferred to K. rhizophila plates. Similar reductions in pgp-5p::gfp expression were observed in animals fed 23 other dsRNAs (Table 1). However, in eft-3(q145);pgp-5p::gfp animals fed lbp-5 dsRNA, we found that pgp-5p::gfp expression was not suppressed ( Figure 4D ). lbp-5 encodes an intracellular fatty acid-binding protein predicted to function as a transporter of hydrophobic molecules such as lipids and steroid hormones ( Xu et al., 2014 ).

[0181] Table 1. Known carotenoid-binding protein genes tested for suppression of gfp expression in eft-3(q145);pgp-5p::gfp animals in response to K. rhizophila. TIFF2026012220000003.tif75158TIFF2026012220000004.tif247158+, GFP on;-, GFP off

[0182] To further identify eukaryotic targets of K. rhizophila carotenoids, we performed pull-down assays using rat liver cell extracts. Liver was chosen for identifying carotenoid-binding proteins for several reasons: first, it is the primary site of xenobiotic detoxification. Second, it is the site of bile acid biosynthesis. Third, the liver has a known carotenoid transport system. Fourth, large amounts of tissue can be easily obtained, which was not possible with C. elegans. To identify carotenoid-binding proteins, protein extracts from rat liver were incubated with K. rhizophila carotenoids and size-exclusion chromatography was used to remove unbound carotenoids. The carotenoid-binding protein extracts were subjected to anion-exchange chromatography, and the protein fractions were eluted. The yellow fractions (indicating the presence of carotenoids) were pooled, concentrated, and desalted. The enriched peaks, yellow-orange in color indicating the presence of carotenoids, were analyzed by native PAGE. The visible yellow-orange bands were excised and subjected to mass spectrometry analysis to identify the proteins. Mass spectrometry identified 48 proteins (Table 2). Interestingly, one of the identified proteins was FABP1 (fatty acid-binding protein 1), a homolog of C. elegans LBP-5. Among the proteins identified by mass spectrometry was PAK2 (p21 protein kinase), a homolog of C. elegans pak-1, which we previously identified in a genome-wide RNAi screen as a hit required for the induction of pgp-5p::gfp in response to translational repression (Govindan et al., 2015). To determine whether any of these 48 proteins were required for the induction of pgp-5p::gfp in eft-3(q145);pgp-5p::gfp, we performed an RNAi screen of the C. elegans homologs of these rat proteins. E. coli expressing dsRNA against the C. elegans homologs of each of the carotenoid-binding proteins was fed to eft-3(q145);pgp-5p::gfp animals, and once these animals reached adulthood, they were screened to determine whether they disrupted GFP induction.In this screen, RNAi of many carotenoid-binding proteins, except for pak-1 RNAi, did not block the induction of pgp-5p::gfp in eft-3(q145);pgp-5p::gfp animals (Table 2). Interestingly, one of the carotenoid-binding proteins identified is AMACR (alpha-methylacyl-CoA racemase), which is required for bile acid biosynthesis (Autio et al., 2014). In C. elegans, C24A3.4 and ZK892.4 are homologs of AMACR (Figure 16A). Although RNAi of C24A3.4 or ZK892.4 individually did not suppress the induction of pgp-5p::gfp in eft-3(q145);pgp-5p::gfp animals, dual RNAi of C24A3.4 and ZK892.4 suppressed the induction of pgp-5p::gfp (Figure 16B).

[0183] To determine whether any of these 48 C. elegans proteins were required for carotenoid-induced suppression of pgp-5p::gfp induction in K. rhizophila, we performed an RNAi screen of the C. elegans homologs of these proteins. When these animals reached adulthood, they were transferred to K. rhizophila-seeded plates and scored for GFP induction 24 hours later. Expression of pgp-5p::gfp was suppressed in animals fed the dsRNA negative control and transferred to K. rhizophila plates (Table 2). A similar reduction in pgp-5p::gfp expression was observed in most dsRNA-fed animals (Table 2); however, pgp-5p::gfp induction was not suppressed in animals treated with chc-1, fcho-1, or lbp-5 RNAi (Figure 4D). chc-1 encodes the C. elegans clathrin heavy chain ortholog, while fcho-1 encodes the C. elegans homolog of the F-BAR domain-containing fer / Cip4 homology domain-only (FCHo) family of proteins. Both CHC-1 and FCHO-1 mediate endocytic trafficking (Grant et al., 2006). An alternative model is that chc-1, fcho-1, or lbp-5 RNAi by itself can induce pgp-5p::gfp expression even in the absence of ribosomal stress; however, RNAi of these genes did not induce pgp-5p::gfp expression (Figure S16C).

[0184] Without wishing to be bound by theory, based on our findings, we propose a model for how K. rhizophila carotenoids inhibit the xenobiotic detoxification response (Figure 4E). Carotenoids released from K. rhizophila enter the C. elegans intestine via clathrin-mediated endocytosis. Within the intestinal cytoplasm, carotenoids released from endocytic vesicles then bind to LBP-5 and are delivered to peroxisomes, where they inhibit bile acid biosynthesis via binding to AMACR. Furthermore, carotenoids also inhibit PAK-1, disrupting the xenobiotic-induced upregulation of the detoxification response.

[0185] (Table 2) List of proteins identified by mass spectrometry and the effect of RNAi of their C. elegans homologs on the suppression of gfp expression in eft-3(q145);pgp-5p::gfp animals in response to K. rhizophila TIFF2026012220000005.tif217155TIFF2026012220000006.tif255155TIFF2026012220000007.tif114155+, GFP on; -, GFP off; blank = not tested

[0186] Example references: TIFF2026012220000008.tif99146TIFF2026012220000009.tif238147TIFF2026012220000010.tif231147 TIFF2026012220000011.tif231147TIFF2026012220000012.tif238147TIFF2026012220000013.tif40140

[0187] Other Aspects While embodiments of the present invention have been described in conjunction with a detailed description thereof, it should be understood that the foregoing description is intended to be illustrative and not to limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the appended claims.

[0188] Sequence information SEQUENCE LISTING <110> THE GENERAL HOSPITAL CORPORATION <120> CAROTENOIDS FOR TREATING OR PREVENTING NAUSEA <150> US 62 / 802,398 <151> 2019-02-07 <160> twenty one <170> PatentIn version 3.5 <210> 1 <211> 464 <212> PRT <213> Arthrobacter arilaitensis <220> <221> MOD_RES <222> (14)..(14) <223> G or R <220> <221> MOD_RES <222> (42)..(42) <223> G or E <220> <221> MOD_RES <222> (132)..(132) <223> A or D <220> <221> MOD_RES <222> (199)..(199) <223> G or D <220> <221> MOD_RES <222> (275)..(275) <223> G or D <220> <221> MOD_RES <222> (375)..(375) <223> G or D <400> 1 Met Asn His Gln Asp Gln Glu Val Val Val Ile Gly Gly Xaa Phe Ser 1 5 10 15 Gly Leu Ala Ser Ala Gly Leu Leu Ala Ala Arg Gly Cys Lys Val Thr 20 25 30 Leu Ile Glu Gln Gln Glu His Pro Gly Xaa Arg Ser Gly Arg Leu Glu 35 40 45 Arg Glu Gly Phe Arg Phe Asp Thr Gly Pro Ser Trp Tyr Leu Met Pro 50 55 60 Glu Val Phe Asp His Trp Phe Arg Leu Met Gly Thr Ser Thr Ala Glu 65 70 75 80 Gln Leu Asp Leu Arg Glu Leu Pro Thr Gly Tyr Arg Val Phe Phe Gln 85 90 95 Asn Arg Gln Ala Pro Ala Asp Phe Gly Ile Gly Ala Ala Ala Ser Asp 100 105 110 Leu Phe Glu Thr Leu Glu Pro Gly Ser Ala Pro Ala Leu Glu Arg Tyr 115 120 125 Leu Arg Thr Xaa Lys Glu Gly Tyr Glu Leu Ala Leu Glu His Phe Leu 130 135 140 Tyr Asp Asp Phe His Ser Leu Lys Ser Leu Leu Asp Pro Arg Ile Leu 145 150 155 160 Arg Arg Ala Pro Gln Leu Ala Lys Leu Leu Ser Thr Ser Leu Gln Ser 165 170 175 His Val Ala Arg Arg Phe Ala Ser Asn Glu Ile Arg Gln Val Leu Gly 180 185 190 Tyr Pro Ala Val Phe Leu Xaa Ser Ser Pro Glu Lys Thr Pro Ala Leu 195 200 205 Tyr Gln Leu Met Ser His Leu Asp Leu Ala Asp Gly Val Lys Tyr Pro 210 215 220 Met Gly Gly Phe Ala Ala Val Ala Asp Ala Met Ala Gln Leu Ala Arg 225 230 235 240 Lys His Gly Ala Gln Ile Gln Leu Gly Ala Thr Ala Thr Ala Ile Glu 245 250 255 Thr Ser Thr Gly Lys Asn Leu Ala Val Ser Ala Val Arg Trp Ile Asp 260 265 270 Ala Asn Xaa Thr Leu His Arg Thr Pro Ala Thr Lys Val Ile Gly Ala 275 280 285 Ala Asp Val Arg His Leu Glu Gly Glu Leu Leu Pro Glu Ser Leu Gln 290 295 300 Thr His Thr Ala Lys Ser Phe Ala Arg Lys Asp Pro Gly Pro Ser Ala 305 310 315 320 Val Leu Leu Cys Leu Gly Ile Lys Gly Lys Leu Pro Gln Leu Glu His 325 330 335 His Asn Leu Leu Phe Thr Glu Asp Trp Ser Glu Asn Phe Ser Arg Ile 340 345 350 Arg Gln Gly Arg Glu Leu Glu Pro Glu Thr Ser Ile Tyr Val Cys Lys 355 360 365 Pro Ser Ala Thr Asp Pro Xaa Thr Ala Pro Glu Gly Cys Glu Asn Leu 370 375 380 Phe Ile Leu Val Pro Ala Pro Ala Leu Pro Glu Trp Gly Ile Gly Ala 385 390 395 400 Ala Asp Gly Gln Gly Asp Ala Ala Val Glu Ala Val Ala Glu Ala Ala 405 410 415 Ile Asp Gln Leu Ser Ala Trp Ala Arg Ile Asp Asp Leu Arg Glu Arg 420 425 430 Ile Val Val Arg Gln Ser Ile Gly Pro Gly Asp Phe Ala Gln Gln Tyr 435 440 445 Gly Ala Tyr Arg Gly Gly Ala Leu Gly Leu Ala His Thr Leu Gly Gln 450 455 460 <210> 2 <211> 572 <212> PRT <213> Kocuria rhizophila <220> <221> MOD_RES <222> (35)..(35) <223> G or R <220> <221> MOD_RES <222> (63)..(63) <223> G or E <220> <221> MOD_RES <222> (154)..(154) <223> G or D <220> <221> MOD_RES <222> (221)..(221) <223> G or D <220> <221> MOD_RES <222> (379)..(379) <223> G or D <220> <221> MOD_RES <222> (478)..(478) <223> G or D <400> 2 Met Thr Asp Gly Asn Arg Thr Thr Arg His Pro Thr Arg Pro Glu Asn 1 5 10 15 Ala His Arg Gly Ser Pro Asp Gly Gly Ala Pro Arg Thr Val Val Val 20 25 30 Gly Gly Xaa Phe Ala Gly Leu Ala Thr Ala Gly Leu Leu Ala Arg Asp 35 40 45 Gly His Arg Val Thr Leu Leu Glu Gln Arg Asp Thr Leu Gly Xaa Arg 50 55 60 Ser Gly Arg Trp Ser Ala Glu Gly Phe Thr Phe Asp Thr Gly Pro Ser 65 70 75 80 Trp Tyr Leu Met Pro Glu Val Ile Asp Arg Trp Phe Thr Leu Met Gly 85 90 95 Ser Ser Ala Asp Glu Gln Leu Asp Leu Arg Arg Leu Asp Pro Gly Tyr 100 105 110 Arg Thr Phe Phe Glu Gln His Leu Asp Glu Pro Pro Thr Asp Val Arg 115 120 125 Ala Gly His Ala Glu Glu Leu Phe Glu Arg Leu Asp Pro Gly Ser Ser 130 135 140 Glu Ala Leu Arg Ala Tyr Leu Gly Ser Xaa Ala Glu Val Tyr Asp Leu 145 150 155 160 Ala Lys Lys His Phe Leu Tyr Thr Asn Phe Ser Arg Pro Thr Asp Leu 165 170 175 Ala Arg Ala Glu Val Leu Arg Asn Leu Pro Arg Leu Gly Gly Leu Leu 180 185 190 Ser Thr Ser Met Gln Arg Tyr Val Ala Ala Arg Phe Arg Asp Pro Arg 195 200 205 Gln Arg Gln Ile Leu Gly Tyr Pro Ala Val Phe Leu Xaa Ala Ser Pro 210 215 220 Asp Thr Ala Pro Ala Met Tyr His Leu Met Ser His Leu Asp Leu Thr 225 230 235 240 Asp Gly Val Gln Tyr Pro Val Gly Gly Phe Ala Ala Leu Val Asp Ala 245 250 255 Met Glu Arg Leu Val Arg Ala Ala Gly Val Glu Ile Val Thr Gly Ala 260 265 270 Glu Val Thr Gly Ile Glu Val Ala Pro Ala Pro Ala Ser Leu Arg Ser 275 280 285 Arg Val Gly Ala Ala Arg Ala Arg Arg Arg Ser Ala Gly Ser Val Thr 290 295 300 Gly Val Thr Trp Arg Ala Ala Ala Pro Glu Glu Gly Val Arg Ala Gly 305 310 315 320 Gln Asp Gly Ala Ala Gly Ala Pro Gly Ala Arg Gly Ala Val Arg Asp 325 330 335 Ala Asp Gly Pro Gly Gly Val Val Glu Gly Pro Gly Val Val Ala Glu 340 345 350 Gly Arg Gly Thr Arg Thr Asp Ala Ser Ala Glu Ala Arg Gly Ala Gly 355 360 365 Thr Asp Ala Pro Ala Gly Gln Pro Gly Asp Xaa Glu Glu Arg Thr Val 370 375 380 Thr Ala Asp Val Val Ile Gly Ala Ala Asp Leu His His Leu Gln Thr 385 390 395 400 Arg Leu Leu Pro Asp Asp Phe Arg Ala Pro Glu Ser Arg Trp Thr His 405 410 415 Arg Asp Pro Gly Pro Ser Gly Val Leu Val Cys Leu Gly Val Arg Gly 420 425 430 Lys Leu Pro Gln Leu Val His His Asn Leu Leu Phe Thr Ala Asp Trp 435 440 445 Asp Asp Asn Phe Gly Arg Ile Ala Asp Gly Thr Pro Leu Ala Glu Gln 450 455 460 Thr Ser Ile Tyr Val Ser Met Thr Ser Ala Thr Asp Pro Xaa Thr Ala 465 470 475 480 Pro Glu Gly Asp Glu Asn Leu Phe Ile Leu Val Pro Ser Pro Ala Val 485 490 495 Pro Glu Trp Gly Arg Gly Gly Val Arg Thr Pro Asp Thr Asp Glu Pro 500 505 510 Gly Ser Pro Gln Val Glu Arg Val Ala Asp Ala Ala Ile Ala Gln Leu 515 520 525 Ala Arg Trp Ala Glu Ile Pro Asp Leu Ala Glu Arg Ile Val Val Arg 530 535 540 Arg Thr Tyr Gly Pro Gly Asp Phe Glu Ala Gln Phe Asn Ala Trp Arg 545 550 555 560 Gly Ser Met Leu Gly Pro Gly His Thr Leu Arg Gln 565 570 <210> 3 <211> 506 <212> PRT <213> Micrococcus luteus <220> <221> MOD_RES <222> (18)..(18) <223> G or R <220> <221> MOD_RES <222> (46)..(46) <223> G or E <220> <221> MOD_RES <222> (137)..(137) <223> A or D <220> <221> MOD_RES <222> (204)..(204) <223> G or D <220> <221> MOD_RES <222> (308)..(308) <223> G or D <220> <221> MOD_RES <222> (407)..(407) <223> G or D <400> 3 Met Ser Ala Arg Asp Thr Ala Leu Gly Pro Arg Thr Val Val Val Gly 1 5 10 15 Gly Xaa Phe Ala Gly Leu Ala Thr Ala Gly Leu Leu Ala Arg Asp Gly 20 25 30 His Arg Val Thr Leu Leu Glu Arg Gly Ala Val Leu Gly Xaa Arg Ala 35 40 45 Gly Arg Trp Ser Glu Ala Gly Phe Thr Phe Asp Thr Gly Pro Ser Trp 50 55 60 Tyr Leu Met Pro Glu Val Ile Asp Arg Trp Phe Arg Leu Met Gly Thr 65 70 75 80 Ser Ala Ala Glu Arg Leu Asp Leu Arg Arg Leu Asp Pro Gly Tyr Arg 85 90 95 Val Tyr Phe Glu Gly His Leu His Glu Pro Pro Val Asp Val Arg Thr 100 105 110 Gly His Ala Glu Thr Leu Phe Glu Ser Leu Glu Pro Gly Ala Gly Arg 115 120 125 Arg Leu Arg Ala Tyr Leu Asp Ser Xaa Ser Arg Ile Tyr Gly Leu Ala 130 135 140 Lys Glu His Phe Leu Tyr Thr Asp Phe Arg Arg Pro Ala Ala Leu Ala 145 150 155 160 His Pro Asp Val Leu Arg Ala Leu Pro Ala Leu Gly Pro Gln Leu Leu 165 170 175 Gly Gly Leu Arg Ser His Val Ala Ala Arg Phe Gln Asp Pro Arg Leu 180 185 190 Arg Gln Ile Leu Gly Tyr Pro Ala Val Phe Leu Xaa Thr Ser Pro Asp 195 200 205 Arg Ala Pro Ala Met Tyr His Leu Met Ser His Leu Asp Leu Ala Asp 210 215 220 Gly Val Gln Tyr Pro Leu Gly Gly Phe Ala Ala Leu Val Asp Ala Met 225 230 235 240 Ala Glu Val Val Arg Glu Ala Gly Val Glu Ile Arg Thr Gly Val Glu 245 250 255 Ala Thr Ala Val Glu Val Ala Asp Arg Pro Ala Pro Ala Gly Arg Leu 260 265 270 Gly Arg Leu Ala Ala Arg Leu Pro Arg Pro Gly Ala Ala Arg Gly Asp 275 280 285 Glu Gly Arg Arg Arg Arg Pro Gly Arg Val Thr Gly Val Ala Trp Arg 290 295 300 Ser Asp Asp Xaa Ala Ala Gly Arg Leu Asp Ala Asp Val Val Val Ala 305 310 315 320 Ala Ala Asp Leu His His Val Gln Thr Arg Leu Leu Pro Pro Gly Arg 325 330 335 Arg Val Ala Glu Ser Thr Trp Asp Arg Arg Asp Pro Gly Pro Ser Gly 340 345 350 Val Leu Val Cys Val Gly Val Arg Gly Ser Leu Pro Gln Leu Ala His 355 360 365 His Thr Leu Leu Phe Thr Ala Asp Trp Glu Asp Asn Phe Gly Arg Ile 370 375 380 Glu Arg Gly Glu Asp Leu Ala Ala Asp Thr Ser Ile Tyr Val Ser Arg 385 390 395 400 Thr Ser Ala Thr Asp Pro Xaa Val Ala Pro Glu Gly Asp Glu Asn Leu 405 410 415 Phe Ile Leu Val Pro Ala Pro Ala Glu Pro Gly Trp Gly Arg Gly Gly 420 425 430 Ile Arg Val Arg Asp Gly Gln Gly Trp Arg Val Asp Arg Ala Gly Asp 435 440 445 Ala Gln Val Glu Ala Val Ala Asp Arg Ala Leu Asp Gln Leu Ala Arg 450 455 460 Trp Ala Gly Ile Pro Asp Leu Ala Glu Arg Ile Val Val Arg Arg Thr 465 470 475 480 Tyr Gly Pro Gly Asp Phe Ala Ala Asp Val His Ala Trp Arg Gly Ser 485 490 495 Leu Leu Gly Pro Gly His Thr Leu Ala Gln 500 505 <210> 4 <211> 479 <212> PRT <213> Corynebacterium glutamicum <220> <221> MOD_RES <222> (20)..(20) <223> G or R <220> <221> MOD_RES <222> (48)..(48) <223> G or E <220> <221> MOD_RES <222> (141)..(141) <223> A or D <220> <221> MOD_RES <222> (208)..(208) <223> S or D <220> <221> MOD_RES <222> (285)..(285) <223> D or absent <220> <221> MOD_RES <222> (389)..(389) <223> G or D <400> 4 Met Lys Val Ser Thr Lys Thr Pro Arg Ser Ser Gly Thr Ala Val Val 1 5 10 15 Ile Gly Ala Xaa Val Ala Gly Leu Ala Thr Ser Ala Leu Leu Ala Arg 20 25 30 Asp Gly Trp Gln Val Thr Val Leu Glu Lys Asn Thr Asp Val Gly Xaa 35 40 45 Arg Ala Gly Ser Leu Glu Ile Ser Gly Phe Pro Gly Phe Arg Trp Asp 50 55 60 Thr Gly Pro Ser Trp Tyr Leu Met Pro Glu Ala Phe Asp His Phe Phe 65 70 75 80 Ala Leu Phe Gly Ala Cys Thr Ser Asp Tyr Leu Asp Leu Val Glu Leu 85 90 95 Thr Pro Gly Tyr Arg Val Phe Ser Gly Thr His Asp Ala Val Asp Val 100 105 110 Pro Thr Gly Arg Glu Glu Ala Ile Ala Leu Phe Glu Ser Ile Glu Pro 115 120 125 Gly Ala Gly Ala Lys Leu Gly Asn Tyr Leu Asp Ser Xaa Ala Asp Ala 130 135 140 Tyr Asp Ile Ala Ile Asp Arg Phe Leu Tyr Asn Asn Phe Ser Thr Leu 145 150 155 160 Gly Pro Leu Leu His Arg Asp Val Leu Thr Arg Ala Gly Arg Leu Phe 165 170 175 Ser Leu Leu Thr Arg Ser Leu Gln Lys Tyr Val Asn Ser Gln Phe Ser 180 185 190 Ser Pro Val Leu Arg Gln Ile Leu Thr Tyr Pro Ala Val Phe Leu Xaa 195 200 205 Ser Arg Pro Thr Thr Thr Pro Ser Met Tyr His Leu Met Ser His Thr 210 215 220 Asp Leu Val Gln Gly Val Lys Tyr Pro Ile Gly Gly Phe Thr Ala Val 225 230 235 240 Val Asn Ala Leu His Gln Leu Ala Leu Glu Asn Gly Val Glu Phe Gln 245 250 255 Leu Asp Ser Glu Val Ile Ser Ile Asn Thr Ala Ser Ser Arg Gly Asn 260 265 270 Thr Ser Ala Thr Gly Val Ser Leu Leu His Asn Arg Xaa Lys Val Gln 275 280 285 Asn Leu Asp Ala Asp Leu Val Val Ser Ala Gly Asp Leu His His Thr 290 295 300 Glu Asn Asn Leu Leu Pro Arg Glu Leu Arg Thr Tyr Pro Glu Arg Tyr 305 310 315 320 Trp Ser Asn Arg Asn Pro Gly Ile Gly Ala Val Leu Ile Leu Leu Gly 325 330 335 Val Lys Gly Glu Leu Pro Gln Leu Asp His His Asn Leu Phe Phe Ser 340 345 350 Glu Asp Trp Thr Asp Asp Phe Ala Val Val Phe Asp Gly Pro Gln Leu 355 360 365 Thr Arg Pro His Asn Ala Ser Asn Ser Ile Tyr Val Ser Lys Pro Ser 370 375 380 Thr Ser Glu Asp Xaa Val Ala Pro Ala Gly Tyr Glu Asn Leu Phe Val 385 390 395 400 Leu Ile Pro Thr Lys Ala Ser Ser Ser Ile Gly His Gly Asp Ala Tyr 405 410 415 Met Gln Ser Ala Ser Ala Ser Val Glu Thr Ile Ala Ser His Ala Ile 420 425 430 Asn Gln Ile Ala Thr Gln Ala Gly Ile Pro Asp Leu Thr Asp Arg Ile 435 440 445 Val Val Lys Arg Thr Ile Gly Pro Ala Asp Phe Glu His Arg Tyr His 450 455 460 Ser Trp Val Gly Ser Ala Leu Gly Pro Ala His Thr Leu Arg Gln 465 470 475 <210> 5 <211> 459 <212> PRT <213> Microbacterium testaceum <220> <221> MOD_RES <222> (12)..(12) <223> G or R <220> <221> MOD_RES <222> (40)..(40) <223> G or E <220> <221> MOD_RES <222> (132)..(132) <223> A or D <220> <221> MOD_RES <222> (199)..(199) <223> G or D <220> <221> MOD_RES <222> (269)..(269) <223> D or absent <220> <221> MOD_RES <222> (370)..(370) <223> S or D <400> 5 Met Ser Ala Gln Arg Ile Val Val Val Gly Gly Xaa Ile Ala Gly Leu 1 5 10 15 Gly Thr Ala Ala Leu Leu Ala Asp Arg Gly His Asp Val His Leu Phe 20 25 30 Glu Ala Arg Asp Ala Leu Gly Xaa Arg Ala Gly Ser Trp Glu Lys Asp 35 40 45 Gly Phe Arg Phe Asp Thr Gly Pro Ser Trp Tyr Leu Met Pro Glu Val 50 55 60 Phe Asp His Phe Phe Arg Leu Leu Gly Thr Ser Ala Ala Glu Gln Leu 65 70 75 80 Asp Leu Val Arg Leu Asp Pro Ala Tyr Arg Val Tyr Gly Pro Pro Gly 85 90 95 Lys Gly Glu Pro Ile Asp Ile Val Ser Gly Arg Glu Ala Val Arg Ala 100 105 110 Leu Phe Glu Lys His Glu Pro Gly Ser Gly Asp Asn Ile Asp Ala Tyr 115 120 125 Leu Asp Ser Xaa Lys Asp Ala Tyr Glu Leu Ser Thr Ser Lys Phe Leu 130 135 140 Tyr Asp Pro Tyr Ser Ser Thr Lys Gly Leu Arg Asp Pro Ala Leu Val 145 150 155 160 Lys Arg Leu Pro Thr Leu Ile Pro Leu Leu Thr Arg Thr Leu Trp Lys 165 170 175 Arg Val Thr Ser Asp Phe Lys Asn Thr Arg Leu Gln Gln Ile Leu Ala 180 185 190 Tyr Pro Ala Val Phe Leu Xaa Gly Ser Pro Phe Glu Val Pro Ser Leu 195 200 205 Tyr His Leu Met Ser His Leu Asp Leu Gly Asp Gly Val Leu Tyr Pro 210 215 220 Lys Gly Gly Met Thr Glu Ile Ile Thr Ala Ile Glu Lys Leu Ala Arg 225 230 235 240 Gly Arg Gly Val Thr Ile Glu Thr Ser Ala Pro Val Glu Ala Ile Ile 245 250 255 Thr Glu Ser Gly Thr Ala Arg Gly Val Arg Leu Ala Xaa Asp Gly Arg 260 265 270 Ile Phe Ala Ala Asp Ala Val Val Ser Gly Ala Asp Leu His His Thr 275 280 285 Glu Asn Glu Leu Leu Glu Glu Lys Asp Arg Gln Tyr Pro Glu Lys Trp 290 295 300 Trp Lys Asp Lys Val Pro Ser Pro Gly Ala Leu Leu Leu Leu Leu Gly 305 310 315 320 Val Thr Gly Glu Leu Pro Gln Leu Thr His His Thr Leu Leu Phe Thr 325 330 335 Asp Asp Trp His Thr Asn Phe Asp Ala Ile Phe Gly Glu Asn Lys Lys 340 345 350 Ile Pro Asp Pro Ala Ser Ile Tyr Ile Cys Arg Pro Ser Ala Ser Asp 355 360 365 Asp Xaa Val Ala Pro Glu Gly His Glu Asn Leu Phe Val Leu Val Pro 370 375 380 Val Pro Ala Asp Pro Asp Ser Gly Arg Gly Gly Val Ser Gly Ala Gly 385 390 395 400 Asp Glu Arg Ile Glu Lys Ala Ala Asp Arg Val Ile Ala Gln Ile Gly 405 410 415 Glu Trp Thr Gly Ile Pro Asp Leu Ala Glu Arg Ile Val Val Arg Lys 420 425 430 Thr Ile Ala Pro Glu Asp Phe Lys Glu Asp Leu His Ala Trp His Gly 435 440 445 Asn Ser Leu Gly Leu Ala His Thr Leu Asn Gln 450 455 <210> 6 <211> 404 <212> PRT <213> Leifsonia xyli <220> <221> MOD_RES <222> (12)..(12) <223> G or R <220> <221> MOD_RES <222> (40)..(40) <223> G or E <220> <221> MOD_RES <222> (131)..(131) <223> A or D <220> <221> MOD_RES <222> (198)..(198) <223> G or D <220> <221> MOD_RES <222> (275)..(275) <223> G or D <220> <221> MOD_RES <222> (375)..(375) <223> G or D <400> 6 Met Thr Ala Pro Arg Ala Val Val Ile Gly Gly Xaa Ile Ala Gly Leu 1 5 10 15 Ala Thr Ala Ala Leu Leu Ala Arg Asp Gly Arg Pro Val Thr Leu Leu 20 25 30 Glu Gln His Gly Thr Leu Gly Xaa Arg Ala Gly Arg Trp Glu Thr Ala 35 40 45 Gly Phe Arg Phe Asp Thr Gly Pro Ser Trp Tyr Leu Met Pro Glu Val 50 55 60 Phe Asp His Phe Phe Arg Leu Leu Gly Thr Ser Ala Ala Glu Gln Leu 65 70 75 80 Asp Leu Val Thr Leu Asp Pro Gly Tyr Arg Val Phe Ala Glu Asp Gly 85 90 95 Arg Arg Pro Leu Asp Ile Arg Ala Ala Gly Ala Ala Asn Arg Ala Leu 100 105 110 Phe Glu Ser Val Glu Ser Gly Ala Gly Ala Ala Leu Asp Arg Tyr Leu 115 120 125 Ala Gly Xaa Arg Glu Thr Tyr Gly Leu Ala Val Asp Arg Phe Leu Tyr 130 135 140 Ser Thr Phe Ala Ser Ile Arg Pro Leu Leu Ser Arg Glu Val Leu Ala 145 150 155 160 Arg Thr Gly Arg Leu Ala Arg Leu Leu Leu Glu Pro Leu Asp Arg Tyr 165 170 175 Ala Ala Arg Cys Val Arg Asp Thr Val Leu Arg Gln Ile Leu Gly Tyr 180 185 190 Pro Ala Val Phe Leu Xaa Thr Ser Pro Asp Arg Ala Pro Ser Leu Tyr 195 200 205 His Leu Met Ser His Leu Asp Leu Asp Asp Gly Val Arg Tyr Pro Val 210 215 220 Gly Gly Phe Ala Thr Leu Ile Asp Arg Ile Val Ala Phe Ala Arg Ala 225 230 235 240 Ala Gly Ala Glu Leu Val Thr Asp Ala Arg Val Thr Gly Ile Arg Thr 245 250 255 Gly Val Gly Gly Arg Arg Ala Ser Ala Phe Gly Val Asp Trp Val Asp 260 265 270 Ala Glu Xaa Arg Ser Arg His Lys His Ala Asp Ile Val Val Ser Ala 275 280 285 Val Asp Arg Arg His Thr Glu Thr Gln Leu Leu Pro Pro Ala Leu Arg 290 295 300 Ser Ser Asp Arg Glu Trp Lys Arg Arg Asp Pro Gly Pro Gly Ala Val 305 310 315 320 Leu Ala Met Leu Gly Val Arg Gly Glu Leu Pro Gln Leu Thr His His 325 330 335 Asn Leu Phe Phe Thr Thr Asp Trp Glu Ala Asn Phe Glu Arg Val Phe 340 345 350 Gly Ala Asp Arg Gly Val Pro Asp Pro Ala Ser Leu Tyr Val Cys Lys 355 360 365 Pro Ser Ala Thr Asp Pro Xaa Val Ala Pro Pro Gly His Glu Asn Leu 370 375 380 Phe Val Leu Val Pro Val Pro Ala Asp Thr Ser Ile Gly Ser Gly Gly 385 390 395 400 Ile Asp Gly Gly <210> 7 <211> 304 <212> PRT <213> Corynebacterium glutamicum <220> <221> MOD_RES <222> (33)..(33) <223> S or F <220> <221> MOD_RES <222> (66)..(66) <223> D or N <220> <221> MOD_RES <222> (204)..(204) <223> R or C <400> 7 Met Thr His Gln Asn Ser Pro Leu Phe Leu Lys Ser Ala Leu Arg Leu 1 5 10 15 Tyr Asn Arg Ala Ser Phe Lys Ala Ser His Lys Val Ile Glu Glu Tyr 20 25 30 Xaa Thr Ser Phe Ser Leu Ser Thr Trp Leu Leu Ser Pro Arg Ile Arg 35 40 45 Asn Asp Ile Arg Asn Leu Tyr Ala Val Val Arg Ile Ala Asp Glu Ile 50 55 60 Val Xaa Gly Thr Ala His Ala Ala Gly Cys Ser Thr Ala Lys Ile Glu 65 70 75 80 Glu Ile Leu Asp Ala Tyr Glu Ile Ala Val Leu Ala Ala Pro Gln Gln 85 90 95 Arg Phe Asn Thr Asp Leu Val Leu Gln Ala Tyr Gly Glu Thr Ala Arg 100 105 110 Arg Cys Asp Phe Glu Gln Glu His Val Ile Ala Phe Phe Ala Ser Met 115 120 125 Arg Lys Asp Leu Lys Ala Asn Thr His Asp Pro Asp Ser Phe Thr Thr 130 135 140 Tyr Val Tyr Gly Ser Ala Glu Val Ile Gly Leu Leu Cys Leu Ser Val 145 150 155 160 Phe Asn Gln Gly Arg Thr Ile Ser Lys Lys Arg Leu Glu Ile Met Gln 165 170 175 Asn Gly Ala Arg Ser Leu Gly Ala Ala Phe Gln Lys Ile Asn Phe Leu 180 185 190 Arg Asp Leu Ala Glu Asp Gln Gln Asn Leu Gly Xaa Phe Tyr Phe Pro 195 200 205 Lys Thr Ser Gln Gly Thr Leu Thr Lys Glu Gln Lys Glu Asp Leu Ile 210 215 220 Ala Asp Ile Arg Gln Asp Leu Ala Ile Ala His Asp Ala Phe Pro Glu 225 230 235 240 Ile Pro Val Gln Ala Arg Ile Gly Val Ile Ser Ala Tyr Leu Leu Phe 245 250 255 Gln Lys Leu Thr Asp Arg Ile Glu Ala Thr Pro Thr Ala Asp Leu Leu 260 265 270 Arg Glu Arg Ile Arg Val Pro Leu His Ile Lys Leu Ser Thr Leu Ala 275 280 285 Arg Ala Thr Met Lys Gly Leu Ser Met Ser Ile Tyr Arg Lys Asn Ser 290 295 300 <210> 8 <211> 289 <212> PRT <213> Microbacterium testaceum <220> <221> MOD_RES <222> (27)..(27) <223> S or F <220> <221> MOD_RES <222> (60)..(60) <223> D or N <220> <221> MOD_RES <222> (199)..(199) <223> R or C <400> 8 Met Ser Val Gly Pro Thr Gly Leu Ala Leu Tyr Ser Arg Thr Ala Asp 1 5 10 15 Asp Ala Ala Ala Ala Val Ile His Arg Tyr Xaa Thr Ser Phe Gly Leu 20 25 30 Ala Ala Arg Leu Leu Gly Ala Arg Pro Arg Pro His Val Arg Asn Ile 35 40 45 Tyr Ala Leu Val Arg Val Ala Asp Glu Ile Val Xaa Gly Pro Ala His 50 55 60 Asp Ala Gly Leu Thr Pro Glu Arg Glu Arg Ala Val Leu Asn Ala Leu 65 70 75 80 Glu Asn Glu Val Met Asp Ala Ile Ala Thr Gly Phe Ser Ala Asn Leu 85 90 95 Val Val His Ala Phe Ala Arg Thr Ala Arg Glu Cys Gly Ile Asp Ala 100 105 110 Asp Leu Ile Ala Pro Phe Phe Ala Ser Met Arg Thr Asp Ile Asp Thr 115 120 125 Ala Glu His Asp Asp Leu Ser His Asp Ala Tyr Val Tyr Gly Ser Ala 130 135 140 Glu Val Val Gly Leu Met Cys Leu Gln Val Phe Leu Asn Ala Gly Met 145 150 155 160 Ser Ala Pro Ala Arg Pro Ala Ala Asp Leu Val Asp Gly Ala Arg Arg 165 170 175 Leu Gly Ala Ala Phe Gln Asp Val Asn Phe Leu Arg Asp Leu Ala Asp 180 185 190 Asp Ala Asp Arg Leu Gly Xaa Asp Tyr Leu Asp Gly Ala Ala Asp Asp 195 200 205 Asp Arg Arg Thr Ala Val Leu Asp Arg Ile Asp Ala Asp Leu Ala Ala 210 215 220 Ala Ala Ser Val Ile Pro His Leu Pro Pro Asp Cys Arg Ala Ala Val 225 230 235 240 Thr Ala Ala His Asp Leu Phe Ala Glu Leu Ser Arg Arg Leu Arg Leu 245 250 255 Ser Pro Ala Gly Ala Pro Arg Val Arg Val Pro Asp Gly Val Lys Ala 260 265 270 Thr Leu Ala Ala Arg Ala Leu Leu Gly Arg Pro Pro Lys Gly Pro Arg 275 280 285 Pro <210> 9 <211> 298 <212> PRT <213> Leifsonia xyli <220> <223> Subspecies xyli <220> <221> MOD_RES <222> (35)..(35) <223> S or F <220> <221> MOD_RES <222> (68)..(68) <223> D or N <220> <221> MOD_RES <222> (206)..(206) <223> R or C <400> 9 Met Thr Arg Thr Glu Thr Ala Ala Ala Ala Gly Pro Pro Thr Asp Leu 1 5 10 15 Ala Leu Tyr Thr Arg Ala Ala His Glu Ser Ala Ala Thr Ile Ile His 20 25 30 Gln Tyr Xaa Thr Ser Phe Gly Met Val Thr Arg Leu Leu Ala Pro Arg 35 40 45 Val Arg Pro Arg Val Glu Asp Val Tyr Ala Leu Val Arg Ile Ala Asp 50 55 60 Glu Ile Val Xaa Gly Ala Ala Ala Glu Ala Gly Leu Asp Leu Ala Asp 65 70 75 80 Gln Arg Ala Leu Leu Asp Ala Leu Glu Ala Asp Thr Glu Arg Ala Met 85 90 95 Arg Thr Gly Tyr Ser Ala Asn Leu Val Val His Ser Phe Ala Ala Thr 100 105 110 Ala Arg Asp Ser Gly Ile Gly Val Ala Leu Thr Arg Pro Phe Phe Ala 115 120 125 Ser Met Arg Arg Asp Leu Ser Leu Val Asp Phe Thr Ala Asp Glu Leu 130 135 140 Arg Glu Tyr Val Tyr Gly Ser Ala Glu Val Val Gly Leu Met Cys Leu 145 150 155 160 Ala Val Phe Leu Thr Asp Ser Pro Val Ala Asp Asp Arg Arg Arg Arg 165 170 175 Leu Glu Ala Gly Ala Arg Arg Leu Gly Ala Ala Phe Gln Lys Ile Asn 180 185 190 Phe Leu Arg Asp Leu Ala Ala Asp Tyr Ala Gly Leu Gly Xaa Ser Tyr 195 200 205 Phe Pro Gly Ile Asp Pro Ala Arg Leu Thr Glu Arg Gln Lys Leu Ala 210 215 220 Leu Val Val Asp Ile Asp Gly Asp Leu Gly Ala Ala Ala Asp Ala Ile 225 230 235 240 Ala Glu Leu Pro Gly Asn Cys Arg Arg Ala Ile Val Ala Ala His Ala 245 250 255 Leu Phe Ser Glu Leu Ser Asp Arg Ile Arg Ala Thr Pro Ala Arg Asp 260 265 270 Leu Ile Val Arg Arg Val Ser Val Pro Met Arg Thr Lys Leu Ala Ile 275 280 285 Leu Leu Arg Ala Thr Ala Gly Ile Leu Arg 290 295 <210> 10 <211> 314 <212> PRT <213> Arthrobacter arilaitensis <220> <221> MOD_RES <222> (35)..(35) <223> S or F <220> <221> MOD_RES <222> (68)..(68) <223> D or N <220> <221> MOD_RES <222> (209)..(209) <223> R or C <400> 10 Met Thr Arg Glu Phe Ser Ser Thr Asp Thr Thr Gly Thr Ala Ala Leu 1 5 10 15 Glu His Tyr Ser Arg Ala Ala Ser Arg Ser Ala Arg Val Val Leu Gly 20 25 30 Glu Tyr Xaa Thr Ser Phe Ser Leu Ala Cys Arg Leu Leu Asp Ala Ser 35 40 45 Ser Ala Gly His Ile Ala Asn Ile Tyr Ala Leu Val Arg Leu Ala Asp 50 55 60 Glu Ile Val Xaa Gly Val Ala Phe Gln Ala Gly Leu Asp Asp Pro Ala 65 70 75 80 Ile Gly Ala Cys Leu Asp Glu Leu Glu Ala Glu Thr Leu Arg Ala Met 85 90 95 Asp Arg Gly Tyr Ser Thr Asn Met Val Val His Ala Phe Ala Ile Thr 100 105 110 Ala Arg Ala Thr Gly Ile Lys Ala Glu Leu Thr Thr Pro Phe Phe Ala 115 120 125 Ser Met Arg Ala Asp Leu Ser Thr Gly Glu His Asp Ala Arg Ser Leu 130 135 140 Gln Glu Tyr Ile Tyr Gly Ser Ala Glu Val Ile Gly Leu Met Cys Leu 145 150 155 160 Gln Val Phe Ala Ala Met Pro Gly Ala Pro Gln Leu Asn Arg Ala Glu 165 170 175 Glu Gln Arg Thr Lys Leu Ala Ala Arg Ser Leu Gly Ala Ala Phe Gln 180 185 190 Lys Val Asn Phe Leu Arg Asp Leu Ala Gln Asp Ser Gln Glu Leu Gly 195 200 205 Xaa Thr Tyr Phe Pro Gly Met Asp Pro Glu Gly Phe Asp Glu Gln Gly 210 215 220 Lys Ala Leu Leu Val Ala Gln Ile Asn Gln Asp Leu Ala Ala Ala Arg 225 230 235 240 Ala Gly Leu Pro Tyr Leu Ala Pro Gln Ala Ala Arg Ala Val Cys Leu 245 250 255 Ala His Asp Leu Phe Gln Glu Leu Asn Val Gln Leu Glu Lys Val Pro 260 265 270 Ala Ala Ala Leu Leu Arg Thr Arg Ile Ser Val Ser Ala Pro Arg Lys 275 280 285 Ala Met Ile Ala Leu Arg Val Leu Leu Gly Ala Gly Thr Pro Ser His 290 295 300 His Lys Leu Arg Met Glu Val Ser Ser Arg 305 310 <210> 11 <211> 303 <212> PRT <213> Kocuria rhizophila <220> <221> MOD_RES <222> (35)..(35) <223> S or F <220> <221> MOD_RES <222> (68)..(68) <223> D or N <220> <221> MOD_RES <222> (209)..(209) <223> R or C <400> 11 Met Arg Thr Pro Met Lys Leu Pro Ala Glu Arg Ala His Thr Pro Leu 1 5 10 15 Arg Leu Tyr Thr Gly Thr Ala Leu Ala Ser Ser Gly Val Val Ile Gly 20 25 30 Glu Tyr Xaa Thr Ser Phe Ser Leu Ala Cys Arg Thr Leu Pro Gly Pro 35 40 45 Val Arg Arg Asp Ile Ala Gly Ile Tyr Ala Leu Val Arg Val Ala Asp 50 55 60 Glu Ile Val Xaa Gly Thr Ala Arg Ala Ala Gly Leu Asp Asp Arg Ala 65 70 75 80 Val Arg Arg Ala Leu Asp Gly Tyr Glu Ala Ala Val Asp Arg Ala Leu 85 90 95 Glu Thr Gly Phe Ser Thr Asp Leu Val Val His Gly Phe Ala Asp Val 100 105 110 Ala Arg Arg His Gly Phe Gly Arg Glu Leu Thr Glu Pro Phe Phe Ala 115 120 125 Ser Met Arg Ala Asp Leu Glu Val Ala Glu His Asp Gly Ala Ser Leu 130 135 140 Glu Asp Tyr Ile Tyr Gly Ser Ala Glu Val Val Gly Leu Met Cys Leu 145 150 155 160 Glu Val Phe Thr Asp Met Pro Gly Thr Arg Ala Gln Thr Pro Glu Gln 165 170 175 Arg Glu Met Leu Arg Ser Thr Ala Arg Arg Leu Gly Ala Ala Phe Gln 180 185 190 Lys Val Asn Phe Leu Arg Asp Leu Gly Ala Asp His Asp Gln Leu Gly 195 200 205 Xaa Thr Tyr Leu Pro Gly Ala Asp Pro Ala His Leu Thr Glu Asp Arg 210 215 220 Lys Ala Ala Leu Leu Ala Asp Leu Asp Ala Asp Leu Asp Ala Ala Val 225 230 235 240 Pro Gly Ile Leu Ala Leu Asp Arg Arg Ala Arg Arg Ala Val Ser Met 245 250 255 Ala His Gly Leu Phe Thr Glu Leu Ala Arg Arg Ile Glu Arg Val Pro 260 265 270 Ala Arg Glu Leu Ser Thr Arg Arg Val Ser Val Pro Thr Ala Val Lys 275 280 285 Leu Gln Ile Ala Ala Arg Ala Ile Ala Asp Thr Glu Val Thr Ala 290 295 300 <210> 12 <211> 298 <212> PRT <213> Micrococcus luteus <220> <221> MOD_RES <222> (28)..(28) <223> S or F <220> <221> MOD_RES <222> (61)..(61) <223> D or N <220> <221> MOD_RES <222> (202)..(202) <223> R or C <400> 12 Met Ala Ala Pro Thr Pro Ser Pro Ala Ala Leu Tyr Thr Arg Thr Ala 1 5 10 15 His Thr Ala Ala Ala Gln Val Ile Arg Arg Tyr Xaa Thr Ser Phe Ser 20 25 30 Trp Ala Cys Arg Thr Leu Pro Arg Gln Ala Arg Gln Asp Val Ala Thr 35 40 45 Ile Tyr Ala Met Val Arg Val Ala Asp Glu Val Val Xaa Gly Val Ala 50 55 60 Val Ala Ala Gly Leu Asp Glu Ala Gly Val Arg Ala Ala Leu Asp Asp 65 70 75 80 Tyr Glu Arg Ala Cys Glu Ala Ala Met Ala Ser Gly Phe Ala Thr Asp 85 90 95 Pro Val Leu His Ala Phe Ala Asp Val Ala Arg Arg His Gly Ile Thr 100 105 110 Pro Glu Leu Thr Arg Pro Phe Phe Ala Ser Met Arg Ala Asp Leu Gly 115 120 125 Ile Arg Glu His Gly Ala Glu Ser Leu Asp Ala Tyr Ile His Gly Ser 130 135 140 Ala Glu Val Val Gly Leu Met Cys Leu Gln Val Phe Leu Ser Leu Pro 145 150 155 160 Gly Thr Arg Ala Arg Thr Pro Gly Gln Arg Gln Glu Leu Arg Ala Gln 165 170 175 Ala Ser Arg Leu Gly Ala Ala Phe Gln Lys Val Asn Phe Leu Arg Asp 180 185 190 Leu Ala Ala Asp His His Glu Leu Gly Xaa Thr Tyr Leu Pro Gly Ala 195 200 205 Ala Pro Gly Val Leu Thr Glu Ala Arg Lys Ala Glu Leu Val Ala Glu 210 215 220 Val Arg Ala Asp Leu Asp Ala Ala Leu Pro Gly Ile Arg Val Leu Asp 225 230 235 240 Pro Gly Ala Gly Arg Ala Val Ala Leu Ala His Gly Leu Phe Ala Ala 245 250 255 Leu Val Asp Arg Ile Glu Ala Thr Pro Ala Ala Glu Leu Ala His Arg 260 265 270 Arg Val Arg Val Pro Asp His Gln Lys Ala Arg Ile Ala Ala Arg Val 275 280 285 Leo Ala Arg Gly Arg Arg Gly Gly Arg Arg 290 295 <210> 13 <211> 287 <212> PRT <213> Corynebacterium glutamicum <220> <221> MOD_RES <222> (139)..(139) <223> D or N <220> <221> MOD_RES <222> (143)..(143) <223> S or F <400> 13 Met Met Glu Lys Ile Arg Leu Ile Leu Leu Ser Ser Arg Pro Ile Ser 1 5 10 15 Trp Ile Asn Thr Ala Tyr Pro Phe Gly Leu Ala Tyr Leu Leu Asn Ala 20 25 30 Gly Glu Ile Asp Trp Leu Phe Trp Leu Gly Ile Val Phe Phe Leu Ile 35 40 45 Pro Tyr Asn Ile Ala Met Tyr Gly Ile Asn Asp Val Phe Asp Tyr Glu 50 55 60 Ser Asp Met Arg Asn Pro Arg Lys Gly Gly Val Glu Gly Ala Val Leu 65 70 75 80 Pro Lys Ser Ser His Ser Thr Leu Leu Trp Ala Ser Ala Ile Ser Thr 85 90 95 Ile Pro Phe Leu Val Ile Leu Phe Ile Phe Gly Thr Trp Met Ser Ser 100 105 110 Leu Trp Leu Thr Leu Ser Val Leu Ala Val Ile Ala Tyr Ser Ala Pro 115 120 125 Lys Leu Arg Phe Lys Glu Arg Pro Phe Ile Xaa Ala Leu Thr Xaa Ser 130 135 140 Thr His Phe Thr Ser Pro Ala Leu Ile Gly Ala Thr Ile Thr Gly Thr 145 150 155 160 Ser Pro Ser Ala Ala Met Trp Ile Ala Leu Gly Ser Phe Phe Leu Trp 165 170 175 Gly Met Ala Ser Gln Ile Leu Gly Ala Val Gln Asp Val Asn Ala Asp 180 185 190 Arg Glu Ala Asn Leu Ser Ser Ile Ala Thr Val Ile Gly Ala Arg Gly 195 200 205 Ala Ile Arg Leu Ser Val Val Leu Tyr Leu Leu Ala Ala Val Leu Val 210 215 220 Thr Thr Leu Pro Asn Pro Ala Trp Ile Ile Gly Ile Ala Ile Leu Thr 225 230 235 240 Tyr Val Phe Asn Ala Ala Arg Phe Trp Asn Ile Thr Asp Ala Ser Cys 245 250 255 Glu Gln Ala Asn Arg Ser Trp Lys Val Phe Leu Trp Leu Asn Tyr Phe 260 265 270 Val Gly Ala Val Ile Thr Ile Leu Leu Ile Ala Ile His Gln Ile 275 280 285 <210> 14 <211> 287 <212> PRT <213> Arthrobacter arilaitensis <220> <221> MOD_RES <222> (136)..(136) <223> D or N <220> <221> MOD_RES <222> (140)..(140) <223> S or F <400> 14 Met Ile Arg Gly Ile Val Ala Ser Ser Arg Pro Ile Ser Trp Val Asn 1 5 10 15 Thr Ala Tyr Pro Phe Ala Ala Ala Tyr Leu Leu Ala Gly Gly Gly Val 20 25 30 Asp Trp Lys Phe Ile Leu Gly Thr Val Phe Phe Leu Phe Pro Tyr Asn 35 40 45 Leu Leu Met Tyr Gly Val Asn Asp Val Phe Asp Tyr Glu Ser Asp Met 50 55 60 Arg Asn Pro Arg Lys Gly Gly Ile Glu Gly Ala Val Leu Ser Lys Gln 65 70 75 80 Ser His Lys Ala Leu Leu Ile Ala Cys Thr Val Cys Ser Leu Pro Phe 85 90 95 Leu Ile Val Leu Ala Ala Gly Gly Asp Ala Ala Ser Asn Ile Thr Leu 100 105 110 Ala Val Ser Ile Phe Ala Val Leu Ala Tyr Ser Ala Pro Arg Leu Arg 115 120 125 Phe Lys Glu Arg Pro Gly Leu Xaa Ser Leu Thr Xaa Ala Val His Phe 130 135 140 Val Ser Pro Ala Val Tyr Gly Trp Val Leu Ala Gly Ser Ala Val Gln 145 150 155 160 Ala Glu Gln Trp Met Val Phe Leu Ala Phe Leu Leu Trp Gly Met Ala 165 170 175 Ser His Ala Leu Gly Ala Ile Gln Asp Ile Ile Pro Asp Arg Gln Gly 180 185 190 Gly Leu Gly Ser Ile Ala Thr Val Leu Ser Ala Arg Lys Thr Ile Tyr 195 200 205 Leu Val Leu Ala Cys Tyr Leu Leu Ala Gly Gly Leu Val Ala Val Gly 210 215 220 Val Ser Gly Ile Gly Arg Trp Ala Ala Val Leu Ser Leu Pro Tyr Val 225 230 235 240 Leu Asn Val Leu Pro His Leu Gly Ile Ser Asp Ala Ser Ser Gly Thr 245 250 255 Val Asn Arg Gly Trp Lys Arg Phe Leu Trp Ile Asn Tyr Leu Cys Gly 260 265 270 Phe Leu Leu Thr Met Leu Leu Ile Phe Ser Ala Phe Phe Thr Tyr 275 280 285 <210> 15 <211> 298 <212> PRT <213> Kocuria rhizophila <220> <221> MOD_RES <222> (147)..(147) <223> D or N <220> <221> MOD_RES <222> (151)..(151) <223> S or F <400> 15 Met Pro Arg Arg Glu Val Asp Val Leu Thr Arg Leu Phe Trp Ala Ser 1 5 10 15 Arg Pro Leu Ser Trp Val Asn Thr Ala Tyr Pro Phe Thr Ala Ala Val 20 25 30 Leu Leu Thr Gly Gly Leu Pro Trp Trp Leu Val Val Leu Gly Thr Val 35 40 45 Phe Phe Leu Val Pro Tyr Asn Leu Ala Met Tyr Gly Ile Asn Asp Val 50 55 60 Phe Asp Tyr Glu Ser Asp Leu Arg Asn Pro Arg Lys Gly Gly Val Glu 65 70 75 80 Gly Ala Val Val Asp Arg Ala Ala Gln Arg Pro Val Leu Leu Ala Ser 85 90 95 Cys Leu Val Pro Ala Pro Phe Val Leu Val Leu Gly Gly Tyr Ala Val 100 105 110 Val Thr Gly Asn Trp Val Ser Ile Ala Val Leu Ala Val Ser Leu Phe 115 120 125 Ala Val Val Ala Tyr Ser Trp Ala Gly Leu Arg Phe Lys Glu Arg Pro 130 135 140 Phe Val Xaa Ala Met Thr Xaa Ala Thr His Phe Val Ser Pro Ala Val 145 150 155 160 Tyr Gly Leu Thr Leu Ala Gly Ala Thr Phe Thr Pro Gly Leu Trp Ala 165 170 175 Leu Leu Ile Gly Phe Phe Leu Trp Gly Met Ala Ser Gln Met Phe Gly 180 185 190 Ala Val Gln Asp Val Val Pro Asp Arg Glu Gly Gly Leu Ala Ser Val 195 200 205 Ala Thr Val Leu Gly Ala Arg Pro Thr Val Trp Val Ala Ala Leu Leu 210 215 220 Tyr Ala Leu Ala Gly Gly Leu Met Val Phe Thr Glu Trp Pro Gly Gln 225 230 235 240 Leu Ala Ala Leu Leu Ala Val Pro Tyr Leu Leu Asn Val Leu Arg Phe 245 250 255 Ser Gly Val Thr Asp Thr Asp Ser Gly Arg Ala Asn Ala Gly Trp Lys 260 265 270 Thr Phe Leu Trp Leu Asn Tyr Leu Thr Gly Phe Leu Val Thr Met Leu 275 280 285 Leu Ile Trp Trp Ala Ala Met Arg Pro Val 290 295 <210> 16 <211> 294 <212> PRT <213> Micrococcus luteus <220> <221> MOD_RES <222> (141)..(141) <223> D or N <220> <221> MOD_RES <222> (145)..(145) <223> S or F <400> 16 Met Ile Arg Thr Leu Phe Trp Val Ser Arg Pro Val Ser Trp Val Asn 1 5 10 15 Thr Ala Tyr Pro Phe Ala Ala Ala Ala Ile Leu Thr Gly Gly Leu Pro 20 25 30 Ala Trp Leu Val Val Leu Gly Val Val Phe Phe Leu Val Pro Tyr Asn 35 40 45 Leu Ala Met Tyr Gly Ile Asn Asp Val Phe Asp Phe Ala Ser Asp Leu 50 55 60 Arg Asn Pro Arg Lys Gly Gly Val Glu Gly Ser Val Leu Gly Asp Pro 65 70 75 80 Ala Val Arg Arg Arg Val Leu Ala Trp Ser Val Leu Leu Pro Val Pro 85 90 95 Phe Val Ala Val Leu Ala Gly Trp Ser Ala Val Arg Gly Glu Trp Ala 100 105 110 Ala Val Leo Val Leo Ala Val Ser Leo Phe Ala Val Val Ala Tyr Ser 115 120 125 Trp Ala Gly Leu Arg Phe Lys Glu Arg Pro Phe Leu Xaa Ala Ala Thr 130 135 140 Xaa Ala Thr His Phe Val Ser Pro Ala Val Tyr Gly Leu Ala Leu Ala 145 150 155 160 Gly Ala Thr Pro Thr Pro Ala Leu Ala Ala Leu Leu Gly Ala Phe Phe 165 170 175 Leu Trp Gly Met Ala Ser Gln Met Phe Gly Ala Val Gln Asp Val Val 180 185 190 Pro Asp Arg Glu Gly Gly Leu Ala Ser Val Ala Thr Val Leu Gly Ala 195 200 205 Arg Arg Thr Val Leu Leu Ala Ala Gly Leu Tyr Ala Ala Ala Gly Leu 210 215 220 Leu Leu Leu Ala Thr Asp Pro Pro Gly Pro Leu Ala Ala Leu Leu Ala 225 230 235 240 Val Pro Tyr Val Val Asn Thr Leu Arg Phe Arg Arg Ile Thr Asp Ala 245 250 255 Thr Ser Gly Ala Ala His Arg Gly Trp Gln Leu Phe Leu Pro Leu Asn 260 265 270 Tyr Val Thr Gly Phe Leu Val Thr Leu Leu Leu Ile Gly Trp Ala Leu 275 280 285 Thr Arg Gly Ala Ala Ala 290 <210> 17 <211> 297 <212> PRT <213> Microbacterium testaceum <220> <221> MOD_RES <222> (147)..(147) <223> D or N <220> <221> MOD_RES <222> (151)..(151) <223> S or F <400> 17 Met Thr Ala Pro Ala Ala Leu Thr Pro Gly Arg Val Leu Arg Glu Leu 1 5 10 15 Phe Val Ser Ser Arg Pro Val Ser Trp Ile Asn Thr Ala Phe Pro Phe 20 25 30 Ala Ala Ala Tyr Leu Leu Thr Thr Arg Gln Ile Asp Ala Thr Leu Ile 35 40 45 Val Gly Ile Leu Phe Phe Leu Val Pro Tyr Asn Leu Ala Met Tyr Gly 50 55 60 Val Asn Asp Val Phe Asp Tyr Glu Ser Asp Leu Arg Asn Pro Arg Lys 65 70 75 80 Gly Gly Thr His Gly Ala Val Leu Asp Lys Arg Met His Pro Ile Thr 85 90 95 Leu Trp Ala Ser Val Leu Ser Cys Leu Pro Phe Val Val Tyr Leu Val 100 105 110 Val Val Gly Ser Pro Leu Ser Trp Leu Val Leu Ala Leu Ser Leu Phe 115 120 125 Phe Val Val Phe Tyr Ser Ala Pro Pro Leu Arg Leu Lys Glu Arg Pro 130 135 140 Phe Ala Xaa Ser Val Thr Xaa Ser Ile His Phe Phe Ser Pro Ala Val 145 150 155 160 Tyr Gly Leu Val Leu Ala Gly Ala Val Trp Thr Trp Gln Leu Val Phe 165 170 175 Val Phe Val Ala Phe Ala Leu Trp Gly Ile Ala Ser His Ala Phe Gly 180 185 190 Ala Val Gln Asp Val Glu Ala Asp Arg Ala Ala Asp Ile Ser Ser Ile 195 200 205 Ala Thr Ala Arg Gly Ala Arg Trp Thr Val Arg Phe Ala Leu Val Ala 210 215 220 Tyr Ala Leu Ala Gly Val Ala Met Leu Phe Thr Ala Trp Pro Gly Pro 225 230 235 240 Leu Ala Gly Val Leu Val Ile Pro Tyr Leu Val Val Cys Trp Pro Tyr 245 250 255 Arg Asn Val Thr Asp Ala Glu Ser Asp Arg Ala Thr Ala Gly Trp Asn 260 265 270 Arg Phe Leu Trp Leu Asn Gln Ile Ala Gly Phe Gly Thr Thr Met Leu 275 280 285 Leu Ile Trp Trp Trp Leu Leu Thr Ala 290 295 <210> 18 <211> 298 <212> PRT <213> Clavibacter michiganensis <220> <221> MOD_RES <222> (148)..(148) <223> D or N <220> <221> MOD_RES <222> (152)..(152) <223> S or F <400> 18 Met Ser Asp Val Arg Ala Arg Pro Gly Ala Ala Glu Met Leu Arg Thr 1 5 10 15 Val Ala Leu Ser Ser Arg Pro Leu Ser Trp Val Asn Thr Ala Phe Pro 20 25 30 Phe Ala Ala Ala Tyr Leu Thr Val Thr Arg Glu Leu Asp Leu Thr Ala 35 40 45 Val Leu Gly Thr Leu Tyr Phe Leu Ile Pro Tyr Asn Leu Ala Met Tyr 50 55 60 Gly Ile Asn Asp Val Phe Asp Tyr Glu Ser Asp Met Arg Asn Pro Arg 65 70 75 80 Lys Gly Gly Val Glu Gly Ala Val Leu Ala Arg Ala Met His Arg Pro 85 90 95 Val Leu Leu Ala Val Leu Val Thr Asn Val Pro Phe Leu Val Tyr Leu 100 105 110 Val Ile Val Gly Ser Ala Ala Ser Ile Ala Val Leu Ala Val Ser Val 115 120 125 Phe Ala Val Ile Ala Tyr Ser Leu Lys Gly Leu Arg Phe Lys Glu Arg 130 135 140 Pro Val Leu Xaa Ser Leu Thr Xaa Ser Thr His Phe Thr Ser Pro Ala 145 150 155 160 Val Tyr Gly Ile Val Leu Ala Gly Gly Ala Phe Thr Pro Ala Leu Trp 165 170 175 Ala Ile Leu Ala Ala Phe Phe Leu Trp Gly Val Ala Ser His Ala Phe 180 185 190 Gly Ala Val Gln Asp Ile Val Ala Asp Arg Glu Gly Gly Ile Ser Ser 195 200 205 Ile Ala Thr Val Leu Gly Gly Ala Val Thr Val Arg Ile Ala Val Leu 210 215 220 Ala Tyr Ala Ala Ala Gly Val Ala Met Leu Phe Thr Gly Leu Pro Gly 225 230 235 240 Ile Ile Ala Ala Val Leu Val Ile Pro Tyr Ile Leu Ser Thr Ala Pro 245 250 255 Phe Trp Ser Ile Arg Asp Glu Asp Ala Gly Ala Ala Asn Arg Gly Trp 260 265 270 Arg Arg Phe Leu Gly Leu Asn Phe Leu Ser Gly Phe Val Val Thr Met 275 280 285 Leu Leu Ile Ala Tyr Trp Leu Thr Thr Ala 290 295 <210> 19 <211> 291 <212> PRT <213> Leifsonia xyli <220> <221> MOD_RES <222> (143)..(143) <223> D or N <220> <221> MOD_RES <222> (147)..(147) <223> S or F <400> 19 Met Ile Val Thr Ala Pro Pro Leu Phe Arg Gln Leu Phe Leu Ala Ser 1 5 10 15 Arg Pro Leu Ser Trp Ile Asn Thr Ala Tyr Pro Phe Ala Ala Ala Tyr 20 25 30 Leu Leu Thr Ala Arg Glu Ala Asp Ile Val Phe Ile Val Gly Thr Leu 35 40 45 Phe Phe Leu Val Pro Tyr Asn Leu Thr Met Tyr Gly Val Asn Asp Val 50 55 60 Phe Asp Tyr Ala Ser Asp Leu Arg Asn Pro Arg Lys Gly Gly Val Glu 65 70 75 80 Gly Ala Leu Leu Asp Pro Gly Thr His Arg Arg Thr Leu Val Ala Ala 85 90 95 Ala Ala Thr Ser Leu Pro Phe Leu Val Phe Leu Ala Leu Ala Gly Pro 100 105 110 Pro Leu Ser Trp Ala Val Leu Ala Gly Ser Leu Phe Phe Val Leu Ala 115 120 125 Tyr Ser Val Arg Gly Leu Arg Phe Lys Lys Val Pro Phe Leu Xaa Ser 130 135 140 Ala Thr Xaa Ser Ile His Phe Val Ser Pro Ala Leu Tyr Gly Leu Val 145 150 155 160 Leu Ala Gly Ala Thr Phe Thr Pro Gly Leu Trp Leu Val Leu Phe Ala 165 170 175 Phe Phe Leu Trp Gly Val Gly Ser His Ala Phe Gly Ala Val Gln Asp 180 185 190 Val Ala Pro Asp Arg Glu Ala Gly Ile Ala Ser Val Ala Thr Val Leu 195 200 205 Gly Ala Ala Arg Thr Val Arg Phe Ala Ile Ala Ala Trp Ala Leu Ala 210 215 220 Ala Leu Ala Ala Leu Ala Ile Pro Trp Pro Gly Pro Leu Val Ala Met 225 230 235 240 Leu Ala Leu Pro Tyr Ile Thr Val Ala Ala Pro Phe Trp Ser Val Pro 245 250 255 Asp Asp Arg Ala Ser Ala Ala Asn Arg Gly Trp Leu Arg Phe Leu Gly 260 265 270 Ile Asn Tyr Ala Cys Gly Cys Leu Leu Thr Leu Leu Leu Ile Gly Tyr 275 280 285 Ala Leu Arg 290 <210> 20 <211> 340 <212> PRT <213> Caenorhabditis elegans <400> 20 Met Tyr Arg Phe Leu Ser Gly Ile Lys Val Val Glu Ile Ala Gly Leu 1 5 10 15 Ala Pro Val Pro His Cys Gly Met Met Leu Ala Asp Phe Gly Ala Asp 20 25 30 Val Thr Val Ile Asp Lys Lys Asn Pro Ala Ile Glu Gln Arg Leu Asn 35 40 45 Arg Gly Lys Thr Met Lys Gln Leu Asp Leu Lys Asn Pro Glu Asp Ile 50 55 60 Lys Lys Val Arg Asp Leu Cys Gln Thr Ser Asp Val Leu Leu Asp Pro 65 70 75 80 Tyr Arg Pro Gly Thr Leu Glu Lys Met Gly Leu Asp Pro Ser Thr Leu 85 90 95 Trp Asn Asn Asn Lys Gly Leu Ile Ile Cys Lys Ile Ser Gly Tyr Gly 100 105 110 Gln Thr Gly Arg Met Ser Gln Glu Thr Gly His Asp Ile Asn Tyr Val 115 120 125 Ala Leu Ser Gly Met Leu Pro Thr Phe Ser Gly Val Asn Ala Thr Arg 130 135 140 Pro Trp Pro Pro Ala Asn Met Leu Ala Asp Phe Ala Gly Gly Gly Leu 145 150 155 160 Ser Ala Ala Phe Gly Ile Leu Ser Ala Ile Tyr Ala Arg Ser His Asn 165 170 175 Gly Gly Lys Gly Cys Leu Leu Asp Cys Ser Met Thr Glu Gly Val Ala 180 185 190 Tyr Leu Ser Ser Phe Val Gln His Tyr Tyr Asp Gln Pro Asn Leu Phe 195 200 205 Thr Asp Lys Tyr Ala Leu Phe Ser Gly Glu Cys Pro Ile Tyr Arg Thr 210 215 220 Tyr Lys Thr Lys Asp Asp Lys Phe Val Ala Val Gly Ala Val Glu Pro 225 230 235 240 Lys Phe Tyr Gln Asn Leu Phe Lys Leu Leu Asn Val Asp Gly Arg Asp 245 250 255 Leu Phe Val Asn Pro Gly Lys Ile Thr Glu Asp Leu Glu Ser Arg Phe 260 265 270 Leu Gln Lys Thr Arg Asp Lys Trp Ala Asn Ile Phe Lys Gly Gln Glu 275 280 285 Cys Cys Val Thr Pro Val Leu Asp Ile His Glu Val Gly Ser Tyr Gly 290 295 300 Gln His Val Asp Arg Asn Ser Phe Thr Lys Thr Ser Ser Asn Trp Ile 305 310 315 320 Ala Asn Pro Ser Pro Arg Val Trp Thr Gln Asp Glu Leu Ala Ala Leu 325 330 335 Ser Ser Lys Lys 340 <210> 21 <211> 343 <212> PRT <213> Caenorhabditis elegans <400> 21 Met Ser Arg Leu Leu Ser Gly Ile Lys Val Val Glu Leu Gly Gly Leu 1 5 10 15 Ala Pro Val Pro Phe Cys Gly Met Ile Leu Ala Asp Phe Gly Ala Asp 20 25 30 Val Thr Val Ile Asp Lys Lys Asn Pro Thr Val Glu Gln Arg Met Asn 35 40 45 Arg Gly Lys Ser Met Lys Glu Phe Asp Leu Arg Lys Ser Glu Asp Ile 50 55 60 Lys Lys Val Arg Asp Leu Cys Arg Thr Ser Asp Val Leu Leu Asp Pro 65 70 75 80 Tyr Arg Pro Gly Thr Leu Glu Lys Met Gly Leu Asp Pro Leu Ser Leu 85 90 95 Trp Asn Asp Asn Lys Gly Leu Ile Ile Cys Arg Ile Ser Gly Tyr Gly 100 105 110 Gln Thr Gly Arg Met Ser Gln Glu Ala Gly His Asp Ile Asn Tyr Val 115 120 125 Ala Met Ser Gly Met Leu Pro Thr Phe Ala Gly Ala Glu Ala Ser Arg 130 135 140 Pro Trp Pro Pro Val Asn Met Leu Ala Asp Phe Ala Gly Gly Gly Leu 145 150 155 160 Ser Ala Ala Phe Gly Ile Val Ser Ala Ile His Ala Arg Thr His Asn 165 170 175 Gly Gly Gln Gly Cys Val Leu Asp Cys Ser Met Thr Glu Gly Val Ala 180 185 190 Tyr Leu Ala Ser Phe Val Gln Tyr Tyr Tyr Glu Gln Ser His Leu Phe 195 200 205 Thr Asp Lys Tyr Ala Ala Phe Thr Gly Glu Cys Pro Ile Tyr Arg Thr 210 215 220 Tyr Lys Thr Lys Asp Gly Lys Phe Met Ala Val Gly Pro Leu Glu Pro 225 230 235 240 Lys Phe His Gln Lys Met Phe Gln Val Leu Gly Val Asn Gly Asp Asp 245 250 255 Leu Phe Ser Glu Pro Glu Arg Ile Thr Lys Val Leu Glu Glu Thr Phe 260 265 270 Leu Gln Lys Thr Arg Asp Glu Trp Ser Ser Ile Phe Glu Gly Gln Asp 275 280 285 Cys Cys Val Thr Pro Val Leu Asp Ile His Glu Val Gly Thr Tyr Gly 290 295 300 Gln His Val Asp Arg Gln Asn Phe Thr Lys Asn Asp Lys Phe Gly Ser 305 310 315 320 Thr Trp Ile Ala Lys Pro Ser Pro Arg Val Lys Thr Pro Glu Glu Leu 325 330 335 Phe Ala Ala Arg Ser Lys Leu 340

Claims

1. administering a therapeutically effective amount of a C50 carotenoid compound to a subject in need thereof.

10. A method for treating nausea and / or vomiting in a subject, comprising:

2. 10. The method of claim 1, wherein the subject has or is at risk of developing nausea and / or vomiting associated with chemotherapy or radiation.

3. 3. The method of claim 1 or 2, wherein the subject has or is at risk of developing chemotherapy-induced nausea and vomiting (CINV) or radiation-induced nausea and vomiting (RINV).

4. 10. The method of claim 1, wherein the subject has or is at risk of developing post-operative nausea and vomiting (PONV).

5. 5. The method of any one of claims 1 to 4, wherein the C50 carotenoid compound is selected from the group consisting of decaprenoxanthin, C50-astaxanthin, C50-β-carotene, C50-carotene (n=3) (16,16-diisopentenylphytoene), C50-zeaxanthin, C50-caloxanthin, C50-nostoxanthin, sarcinaxanthin, sarprenoxanthin, the acyclic C50 carotenoid bacterioruberin, C50-canthaxanthin, C50-lycopene, C50-phytoene, and combinations thereof.

6. 6. The method of claim 5, wherein the C50 carotenoid compound is decaprenoxanthin.

7. The method of any one of claims 1 to 6, wherein the administering step comprises administering (i) a microorganism that synthesizes C50 carotenoid compounds or a component thereof, (ii) an extract of a microorganism that synthesizes C50 carotenoid compounds, (iii) an extracted C50-carotenoid compound, or (iv) a composition that is a combination thereof; or a composition comprising any of them.

8. 8. The method of claim 7, wherein the microorganism that synthesizes the C50 carotenoid compound is viable or alive.

9. 9. The method of claim 8, wherein the administering step comprises administering a sufficient amount of the microorganism to colonize the microbiome of the subject.

10. The method of any one of claims 7 to 9, wherein the composition comprises or is prepared from a culture of the microorganism.

11. 11. The method of claim 10, wherein the microorganism is a strain found in nature.

12. 11. The method of any one of claims 7 to 9 or 10, wherein the microorganism is an engineered microorganism.

13. 13. The method of claim 12, wherein the engineered microorganism comprises a genetic mutation relative to an otherwise identical reference microorganism such that the engineered microorganism produces the C50 carotenoid compound at an absolute or relative level that differs from that of the reference microorganism.

14. 14. The method of any one of claims 1 to 13, wherein said administering step comprises administering a composition comprising or delivering a synthetic C50 carotenoid compound.

15. 8. The method of claim 7, wherein the microorganism that synthesizes the C50 carotenoid compound is selected from the group consisting of Kocuria rhizophila, Corynebacterium glutamicum, Arthrobacter arilaitensis, and combinations thereof.

16. administering a therapeutically effective amount of a C50 carotenoid compound to a subject in need thereof.

11. A method for reducing food aversions in a subject, comprising:

17. 17. The method of claim 16, wherein the subject has or is at risk of developing nausea and vomiting associated with chemotherapy or radiation.

18. 18. The method of claim 16 or 17, wherein the subject has or is at risk of developing chemotherapy-induced nausea and vomiting (CINV) or radiation-induced nausea and vomiting (RINV).

19. 17. The method of claim 16, wherein the subject has or is at risk of developing post-operative nausea and vomiting (PONV).

20. 20. The method of any one of claims 16 to 19, wherein the C50 carotenoid compound is selected from the group consisting of decaprenoxanthin, C50-astaxanthin, C50-β-carotene, C50-carotene (n=3) (16,16-diisopentenylphytoene), C50-zeaxanthin, C50-caloxanthin, C50-nostoxanthin, sarcinaxanthin, sarprenoxanthin, the acyclic C50 carotenoid bacterioruberin, C50-canthaxanthin, C50-lycopene, C50-phytoene, and combinations thereof.

21. 21. The method of claim 20, wherein the C50 carotenoid compound is decaprenoxanthin.

22. The method of any one of claims 16 to 21, wherein the administering step comprises administering (i) a microorganism that synthesizes C50 carotenoid compounds or a component thereof, (ii) an extract of a microorganism that synthesizes C50 carotenoid compounds, (iii) an extracted C50-carotenoid compound, or (iv) a composition that is a combination thereof; or a composition comprising any of them.

23. 23. The method of claim 22, wherein the microorganism that synthesizes the C50 carotenoid compound is viable or living.

24. 24. The method of claim 23, wherein said administering step comprises administering a sufficient amount of said microorganism to colonize the microbiome of said subject.

25. The method of any one of claims 22 to 24, wherein the composition comprises or is prepared from a culture of the microorganism.

26. 26. The method of claim 25, wherein the microorganism is a strain found in nature.

27. 25. The method of any one of claims 22 to 24, wherein the microorganism is an engineered microorganism.

28. 28. The method of claim 27, wherein the engineered microorganism comprises a genetic mutation relative to an otherwise identical reference microorganism such that the engineered microorganism produces the C50 carotenoid compound at an absolute or relative level that differs from that of the reference microorganism.

29. 29. The method of any one of claims 16-28, wherein said administering step comprises administering a composition comprising or delivering a synthetic C50 carotenoid compound.

30. 23. The method of claim 22, wherein the microorganism that synthesizes the C50 carotenoid compound is selected from the group consisting of Kocuria rhizophila, Corynebacterium glutamicum, Arthrobacter alilactensis, and combinations thereof.

31. A therapeutic composition for oral delivery comprising a therapeutically effective amount of a C50 carotenoid compound and a pharmaceutically acceptable carrier.

32. 32. The therapeutic composition of claim 31, comprising a microorganism that synthesizes said C50 carotenoid compound.

33. 33. The therapeutic composition of claim 32, wherein the microorganism is a cultured microorganism.

34. 34. The therapeutic composition of claim 33, wherein the microorganism is an engineered microorganism.

35. 35. The therapeutic composition of claim 34, wherein the engineered microorganism comprises a genetic mutation relative to an otherwise identical reference microorganism such that the engineered microorganism produces the C50 carotenoid compound at an absolute or relative level that differs from that of the reference microorganism.

36. 35. The therapeutic composition of any one of claims 31-34, which is a liquid, syrup, tablet, lozenge, gummy, capsule, powder, gel, or film.

37. 37. The therapeutic composition of any one of claims 31 to 36, wherein said C50 carotenoid compound is at least 20% w / w of said composition.

38. 38. The therapeutic composition of any one of claims 31 to 37, wherein said C50 carotenoid compound is purified.

39. 39. The therapeutic composition of any one of claims 31 to 38, wherein said C50 carotenoid compound has a chemical structure found in nature.

40. 40. The therapeutic composition of any one of claims 31 to 39, wherein said C50 carotenoid compound is an analog of a reference C50 carotenoid compound found in nature.

41. 41. The therapeutic composition of any one of claims 31 to 40, wherein the C50 carotenoid compound is selected from the group consisting of decaprenoxanthin, C50-astaxanthin, C50-β-carotene, C50-carotene (n=3) (16,16-diisopentenylphytoene), C50-zeaxanthin, C50-caloxanthin, C50-nostoxanthin, sarcinaxanthin, sarprenoxanthin, the acyclic C50 carotenoid bacterioruberin, C50-canthaxanthin, C50-lycopene, C50-phytoene, and combinations thereof.

42. 42. The therapeutic composition of claim 41, wherein said C50 carotenoid compound is decaprenoxanthin.

43. 43. The therapeutic composition of any one of claims 31 to 42, wherein said microorganism is live or viable.

44. 44. The therapeutic composition of any one of claims 31 to 43, wherein said microorganism is killed.

45. 45. The therapeutic composition of any one of claims 31-44, wherein the microorganism is selected from the group consisting of Kocuria rhizophila, Corynebacterium glutamicum, Arthrobacter alilactensis, and combinations thereof.

46. A method for producing a therapeutic composition according to any one of claims 31 to 45, comprising the steps of: combining a pharmaceutically acceptable carrier with a C50 carotenoid compound; and formulating the combination into a therapeutic composition.

47. 47. The method of claim 46, wherein said combining step comprises combining a pharmaceutically acceptable carrier with a microorganism that synthesizes a C50 carotenoid compound.

48. 47. The method of claim 46, wherein said combining step comprises combining a pharmaceutically acceptable carrier with a chemically synthesized C50 carotenoid compound.

49. A method for treating nausea and / or vomiting in a subject, comprising: Administering (i) a microorganism that synthesizes C50 carotenoid compounds or a component thereof, (ii) an extract of a microorganism that synthesizes C50 carotenoid compounds, (iii) the extracted C50-carotenoid compounds, or (iv) a combination thereof to a subject in need thereof.

50. 50. The method of claim 49, wherein the subject has or is at risk of developing nausea and / or vomiting associated with chemotherapy or radiation.

51. 51. The method of claim 49 or 50, wherein the subject has or is at risk of developing chemotherapy-induced nausea and vomiting (CINV) or radiation-induced nausea and vomiting (RINV).

52. 50. The method of claim 49, wherein the subject has or is at risk of developing post-operative nausea and vomiting (PONV).

53. 53. The method of any one of claims 49 to 52, wherein the microorganism that synthesizes the C50 carotenoid compound is selected from the group consisting of Kocuria rhizophila, Corynebacterium glutamicum, Arthrobacter alilactensis, and combinations thereof.

54. A method for reducing food aversions in a subject, comprising: Administering (i) a microorganism that synthesizes C50 carotenoid compounds or a component thereof, (ii) an extract of a microorganism that synthesizes C50 carotenoid compounds, (iii) the extracted C50-carotenoid compounds, or (iv) a combination thereof to a subject in need thereof.

55. 55. The method of claim 54, wherein the subject has or is at risk of developing nausea and vomiting associated with chemotherapy or radiation.

56. 56. The method of claim 54 or 55, wherein the subject has or is at risk of developing chemotherapy-induced nausea and vomiting (CINV) or radiation-induced nausea and vomiting (RINV).

57. 55. The method of claim 54, wherein the subject has or is at risk of developing post-operative nausea and vomiting (PONV).

58. 58. The method of any one of claims 54 to 57, wherein the microorganism that synthesizes the C50 carotenoid compound is selected from the group consisting of Kocuria rhizophila, Corynebacterium glutamicum, Arthrobacter alilactensis, and combinations thereof.

59. Use of a C50 carotenoid compound for treating nausea and / or vomiting in a subject in need thereof.

60. Use of a microorganism that synthesizes a C50 carotenoid compound to treat nausea and / or vomiting in a subject in need thereof.

61. 61. The use of claim 60, wherein the microorganism that synthesizes the C50 carotenoid compound is selected from the group consisting of Kocuria rhizophila, Corynebacterium glutamicum, Arthrobacter alilactensis, and combinations thereof.

62. Use of a C50 carotenoid compound to reduce food aversion in a subject.

63. Use of a microorganism that synthesizes C50 carotenoid compounds to reduce food aversions in a subject.

64. 64. The use of claim 63, wherein the microorganism that synthesizes the C50 carotenoid compound is selected from the group consisting of Kocuria rhizophila, Corynebacterium glutamicum, Arthrobacter alilactensis, and combinations thereof.

65. A method for evaluating carotenoid compounds for anti-nausea and / or anti-vomiting activity, comprising the steps of: (i) contacting the system with a carotenoid compound; (ii) determining whether the carotenoid compound alters a characteristic of the system, the characteristic being associated with nausea and / or vomiting.

66. 66. The method of claim 65, wherein said determining step comprises comparing said characteristics before and after performing said contacting step.

67. 66. The method of claim 65, wherein said determining step comprises, after said contacting step, comparing said characteristic to an equivalent reference.

68. 68. The method of claim 67, wherein the equivalent reference is a historical reference.

69. 68. The method of claim 67, wherein said equivalent reference is a negative control reference.

70. 68. The method of claim 67, wherein said equivalent reference is a positive control reference.

71. 71. The method of any one of claims 65 to 70, wherein the system is or comprises C. elegans.

72. 72. The method of any one of claims 65 to 71, wherein said characteristic is level of food aversion.

73. 72. The method of any one of claims 65 to 71, wherein said characteristic is a level or activity of a nucleic acid or protein, or a form thereof.

74. 74. The method of claim 73, wherein said characteristic is or comprises an aspect of a xenobiotic detoxification response.

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