Fusion proteins / peptides, their production methods and uses

Abstract

Fusion proteins / peptides and biopolymers containing keratin and keratin-related peptide sequences are provided herein. The fusion proteins and biopolymers may further comprise elastin-like, silk-like, collagen or collagen-like, resilin-like, abductin-like peptides, and / or linker-like sequences. The fusion proteins and biopolymers may exhibit functional properties derived from the various peptide sequences incorporated therein, such as elasticity, moisture retention, recovery, texture, or antibacterial and UV protection. Additionally, methods for obtaining and characterizing fusion proteins and biopolymers containing keratin and keratin-related peptide sequences are provided herein. Fusion proteins and biopolymers containing keratin and keratin-related peptide sequences can be included in compositions for improving the properties of hair, skin, and nails.

Description

[Technical Field]

[0001] The present disclosure relates to fusion proteins / peptides with binding modules and biopolymers for hair and skin care, namely fusion proteins and biopolymers with keratin-binding modules.

[0002] The present disclosure relates to fusion proteins and peptides designed for use in hair and skin applications, including cosmetic and dermatological formulations. [Background technology]

[0003] A fusion protein or chimeric protein is a protein created by joining two or more genes encoding originally separate proteins, peptides, or modules. Translation of this fusion gene results in a single or multiple polypeptides that possess the functional properties of each of the original sequences. Some fusion proteins combine full-length peptides and therefore contain all the functional domains of the original proteins. However, other fusion proteins, especially those that occur naturally, combine only parts of the coding sequences and therefore do not maintain the original function of the parent genes that formed them. Novel recombinant technologies have enabled improved fusion protein design for use in diverse fields such as biodetection, agriculture, the paper and food industries, and biopharmaceuticals. Summary of the Invention

[0004] Proteins, peptides, and peptide components or blocks, such as silk and collagen, have been used as ingredients in hair, skin, and nail care compositions. In some cases, they are used to strengthen and moisturize hair, skin, and nails. In other cases, they are used to improve the appearance of hair, skin, and nails and protect them from environmental or chemical damage. However, they may not always perform the desired function when used in hair, skin, and nail care products. For example, the isoelectric point of many natural proteins is lower than desired, so the protein's affinity for negatively charged hair, skin, and nails is not strong enough. Furthermore, many proteins have low solubility due to their high molecular weight and hydrophobicity. Structural proteins, such as silk and collagen, can be chemically degraded, resulting in a mixture of components / blocks of various molecular weights with variable properties. To increase solubility for inclusion in products, these proteins are often chemically modified. However, chemical modification of these proteins can affect desired properties. Often, multiple proteins may be required to achieve all desired properties, which can increase product cost and production time. As a result, there remains an unmet need for designer proteins that overcome the inherent limitations of naturally derived products and could provide better compositions for hair, skin, and nail products.

[0005] These facts are disclosed to explain the technical problem that the present disclosure aims to solve.

[0006] The present disclosure relates to a novel class of novel fusion proteins and peptides specifically designed for optimal performance in hair, nail, and skin applications, i.e., cosmetic and skin formulations. The fusion peptides / proteins of the present disclosure demonstrate remarkable efficacy for a variety of cosmetic and skin-related challenges. The present disclosure relates to these novel fusion peptides / proteins, their synthesis, and uses in hair, nail, or skin care.

[0007] Further aspects and advantages of the present disclosure will become readily apparent to those skilled in the art from the following description, wherein only illustrative embodiments of the present disclosure are shown and described. It will be understood that the present disclosure is capable of other and different embodiments, and its details are capable of modifications in various obvious respects, all without departing from the disclosure.

[0008] The present disclosure relates to fusion proteins comprising keratin peptides and biopolymers comprising keratin peptides for hair and skin care.

[0009] The present disclosure relates to cosmetic and personal care compositions, and more specifically to novel fusion proteins having binding modules designed to improve the affinity, binding properties, and strength of hair, nails, and / or skin. In some embodiments, the hair strengthening effect of the fusion protein is determined by Young's modulus. In some cases, the hair strengthening effect is determined as the relative difference in Young's modulus between hair treated with the fusion protein / peptide or a composition comprising the fusion protein / peptide of the present disclosure and untreated hair. In some cases, treating hair with the fusion protein / peptide improves hair integrity. In some cases, hair integrity is determined by differential scanning calorimetry (DSC).

[0010] In some aspects, the present disclosure relates to a fusion protein / peptide comprising: (a) a keratin or keratin-related peptide block; and (b) at least one polypeptide block selected from the list consisting of: (i) an elastin or elastin-like polypeptide (ELP) block, (ii) a silk or silk-like polypeptide (SLP) block, (iii) a collagen or collagen-like polypeptide (CLP) block, (iv) a resilin or resilin-like polypeptide (RLP) block, (v) an abductin or abductin-like polypeptide (ALP) block, (vi) a linker component as a spacer, or (vii) a combination of two or more selected from (i) to (vi).

[0011] In the present disclosure, the keratin or keratin-associated peptide block may be selected from or may be a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO. 8, SEQ ID NO. 9, SEQ ID NO. 10, SEQ ID NO. 48, SEQ ID NO. 66, SEQ ID NO. 67, SEQ ID NO. 68, SEQ ID NO. 69, SEQ ID NO. 101, SEQ ID NO. 102, SEQ ID NO. 103, and SEQ ID NOs. 70-81.

[0012] In the present disclosure, the elastin or elastin-like polypeptide (ELP) block may be selected from or may be a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NOs. 11-23, 49-55, 82-91, and 104.

[0013] In the present disclosure, the silk or silk-like polypeptide (SLP) block may be selected from or may be a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NOs. 24-32, SEQ ID NO. 56, SEQ ID NO. 105, and SEQ ID NO. 92.

[0014] In the present disclosure, the collagen or collagen-like polypeptide (CLP) block may be selected from or may be a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO. 33, SEQ ID NO. 57-59, SEQ ID NO. 93-95, and SEQ ID NO. 106.

[0015] In the present disclosure, the resilin or resilin-like polypeptide (RLP) block may be selected from or may be a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NOs. 34-35, SEQ ID NO. 60, SEQ ID NO. 96, and SEQ ID NO. 107.

[0016] In the present disclosure, the abductin or abductin-like polypeptide (ALP) block may be or may be selected from sequences with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NOs. 36-42, 61, and 108. In the present disclosure, the linker component, as a spacer, may be or may be selected from sequences with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the following list: SEQ ID NOs. 97-100.

[0017] In embodiments for better results, the polypeptide block may comprise an elastin or elastin-like polypeptide (ELP) block.

[0018] In embodiments for better results, the elastin or elastin-like polypeptide block comprises 8 to 30 amino acids, preferably 10 to 30 amino acids, more preferably 15 to 30 amino acids.

[0019] In an embodiment for better results, the polypeptide block may comprise a linker block as a spacer.

[0020] In embodiments for better results, the linker block comprises a hydrophilic block comprising 1 to 75 amino acids, preferably 10 to 50 amino acids. In some embodiments, the hydrophilic block comprises hydrophilic amino acids. In some embodiments, the hydrophilic amino acids comprise serine, threonine, glutamine, asparagine, glutamic acid, or aspartic acid.

[0021] In embodiments for better results, the linker block adopts an α-helical structure. In some embodiments, the linker block adopts a random coil structure. In some embodiments, the secondary structure of the fusion peptide is determined by circular dichroism (CD) spectroscopy.

[0022] In an embodiment for better results, the linker block comprises 1 to 3 amino acids repeated 0 to 25 times, preferably 1 to 20 times.

[0023] In an embodiment for obtaining better results, the linker block has 1 to 2 or 10 to 25 times.

[0024] In an embodiment for better results, the keratin or keratin-associated peptide block comprises SEQ ID NO:8:GGVCGPSPPCITT or a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO:8.

[0025] In an embodiment for better results, the keratin or keratin-associated peptide block comprises SEQ ID NO:9:GGVCGPSPPC or a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO:9.

[0026] In embodiments for better results, the keratin or keratin-associated peptide block comprises SEQ ID NO: 10:CGPSPPCITT or a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 10.

[0027] In an embodiment for better results, the keratin or keratin-associated peptide block comprises SEQ ID NO:68:CLPCLPAASC or a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO:68.

[0028] In an embodiment for better results, the keratin or keratin-associated peptide block comprises SEQ ID NO: 73:CVSSCCKPQCC or a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 73.

[0029] In an embodiment for better results, the keratin or keratin-associated peptide block comprises SEQ ID NO:74:PITCRRTCYH or a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO:74.

[0030] In an embodiment for better results, the keratin or keratin-associated peptide block comprises SEQ ID NO: 75:DCKLPCNPCA or a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 75.

[0031] In an embodiment for better results, the keratin or keratin-associated peptide block comprises SEQ ID NO: 76:CLPCLPAASC or a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 76.

[0032] In an embodiment for better results, the keratin or keratin-associated peptide block comprises SEQ ID NO:77:CEPAICEPSC or a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO:77.

[0033] In embodiments for better results, the fusion protein / peptide comprises SEQ ID NO.1:GGVCGPSPPCITTVPGVGVPGVGVPGLGVPGVGVPGVGR, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO:1.

[0034] In an embodiment for achieving better results, the fusion protein comprises SEQ ID NO.2:GGVCGPSPPCITTVPGVGVPGVGVPGVGVPGVGVPGVGVPGVG, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO:2.

[0035] In an embodiment for better results, the fusion protein comprises SEQ ID NO.7:GGVCGPSPPCITTVPGVGVPGVGVPGLGVPGVGVPGVG, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO:7.

[0036] In embodiments for better results, the polypeptide block comprises a silk or silk-like polypeptide (SLP) block.

[0037] In some embodiments, the fusion protein comprises SEQ ID NO:3:GGVCGPSPPCITTGAGAGSGAGAGSGAGAGSGAGAGSGAGAGS, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO:3.

[0038] In some embodiments, the polypeptide block comprises a collagen or collagen-like polypeptide (CLP) block.

[0039] In some embodiments, the fusion protein comprises SEQ ID NO:5:GGVCGPSPPCITTGPTGPTGPAGPRGLQGLQGLQGERGEQGPT, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO:5.

[0040] In some embodiments, the polypeptide block comprises a resilin or resilin-like polypeptide (RLP) block.

[0041] In some embodiments, the fusion protein comprises SEQ ID NO:4:GGVCGPSPPCITTGGRPSDSYGAPGGGN, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO:4.

[0042] In some embodiments, the polypeptide block comprises an abductin or abductin polypeptide (ALP) block.

[0043] In some embodiments, the fusion protein comprises SEQ ID NO.6:GGVCGPSPPCITTGGFGGMGGGSGGFGGMGGGSGGFGGMGGGS, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO:6.

[0044] In a specific embodiment, the fusion protein / peptide has the sequence GGVCGPSPPCITTGGFGGMGGGSGGFGGMGGGSGGFGGMGGGS (SEQ ID NO:6).

[0045] In a particular embodiment, the fusion protein / peptide has the sequence GVGVPGVGVPGLGVPGVGVPGVG (SEQ ID NO:22).

[0046] In certain embodiments, the fusion protein / peptide has the sequence GAGAGSGAGAGSGAGAGSGAGAGSGAGAGS (SEQ ID NO:32).

[0047] In a particular embodiment, the fusion protein / peptide has the sequence GPTGPTGPAGPRGLQGLQGLQGERGEQGPT (SEQ ID NO:33).

[0048] In a particular embodiment, the fusion protein / peptide has the sequence GGVCGPSPPCITTGQGQGQGQGQGQGQGQGQGQGQGQ (SEQ ID NO:44).

[0049] In a particular embodiment, the fusion protein / peptide has the sequence GGVCGPSPPCITTAKAKAKAKAKAKAKAKAKAKAKAKAK (SEQ ID NO:45).

[0050] In a particular embodiment, the fusion protein / peptide has the sequence GGVCGPSPPCITTLKLKLKLKLKLKLKLKLKLKLKLK (SEQ ID NO:46).

[0051] In a specific embodiment, the fusion protein / peptide has the sequence GGVCGPSPPCITTGAGAGAGAGAGAGAGAGAGAGAGA (SEQ ID NO:47).

[0052] In certain embodiments, the linker block comprises a spacer-like repeat sequence block. The linker may comprise about 1 to about 3 amino acids repeated 0 to 25 times. The linker may comprise 1 to 2 amino acids repeated 10 to 25 times.

[0053] In certain embodiments, the present disclosure provides GGVCGPSPPCITTVPGVGVPGVGVPGLGVPGVGVPGVGR(SEQ ID NO:1); GGVCGPSPPCITTVPGVGVPGVGVPGVGVPGVGVPGVGVPGVG(SEQ ID NO:2); GGVCGPSPPCITTTGAGAGSGAGAGSGAGAGSGAGAGSGAGAGS(SEQ ID NO:3); GGVCGPSPPCITTGGRPSDSYGAPGGGN(SEQ ID NO:4); GGVCGPSPPCITTGPTGPTGPAGPRGLQGLQGLQGERGEQGPT(SEQ ID NO:5); GGVCGPSPPCITTGGFGGMGGGSGGFGGMGGGSGGFGGMGGGS (SEQ ID NO: 6), or GGVCGPSPPCITTVPGVGVPGVGVPGLGVPGVGVPGVG(SEQ ID NO:7); GGVCGPSPPCITTGQGQGQGQGQGQGQGQGQGQGQGQGQ(SEQ ID NO:44); GGVCGPSPPCITTAKAKAKAKAKAKAKAKAKAKAKAKAK(SEQ ID NO:45); GGVCGPSPPCITTLKLKLKLKLKLKLKLKLKLKLKLK(SEQ ID NO:46); GGVCGPSPPCITTTGAGAGAGAGAGAGAGAGAGAGAGAGA(SEQ ID NO:47) The present invention provides fusion proteins having at least 90% sequence identity to any of the sequences of

[0054] Fusion proteins according to embodiments of the present disclosure comprise keratin or keratin-related peptide blocks having a length of 6-16 amino acids, silk or "SLP" having a repeat sequence having a length of 6-8 amino acids, and / or silk or SLP having a length of 15-30 amino acids, collagen or "CLP" having a repeat sequence having a length of 3 amino acids, and / or collagen or CLP having a length of 15-30 amino acids, elastin or "ELP" having a repeat sequence having a length of 5 amino acids, and / or elastin or ELP having a length of 15-30 amino acids, resilin or "RLP" having a repeat sequence having a length of 11-15 amino acids, and / or resilin or RLP having a length of 15-30 amino acids, abductin or "ALP" having a repeat sequence having a length of 4-10 amino acids, and / or abductin or ALP having a length of 15-30 amino acids.

[0055] In some embodiments, the fusion protein has an average molecular weight of about 1 kDa to about 144 kDa. In some embodiments, the fusion protein has an average molecular weight of about 10 kDa to about 144 kDa. In some embodiments, the fusion protein has an average molecular weight of about 35 kDa to about 130 kDa.

[0056] In some embodiments, the fusion protein is from about 1 kDa to about 5 kDa, from about 5 kDa to about 10 kDa, from about 6 kDa to about 17 kDa, from about 10 kDa to about 15 kDa, from about 14 kDa to about 30 kDa, from about 15 kDa to about 20 kDa, from about 17 kDa to about 39 kDa, from about 20 kDa to about 25 kDa, from about 25 kDa to about 30 kDa, or from about 30 kDa. The average molecular weight ranges are about 1 to about 35 kDa, about 35 kDa to about 40 kDa, about 39 kDa to about 54 kDa, about 39 kDa to about 80 kDa, about 40 kDa to about 45 kDa, about 45 kDa to about 50 kDa, about 50 kDa to about 55 kDa, about 55 kDa to about 60 kDa, about 60 kDa to about 100 kDa, and about 80 kDa to about 144 kDa.

[0057] In some embodiments, the fusion protein comprises a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 1-7, 44-47, or 200-552.

[0058] In some embodiments, the fusion protein comprises a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID NOs: 1-7, 44-47, or 200-552.

[0059] In some embodiments, the fusion protein comprises a sequence identical to any one of SEQ ID NOs: 1-7, 44-47, or 200-552.

[0060] In some aspects, provided herein are compositions comprising fusion proteins according to embodiments of the present disclosure for improving the elasticity, recovery, moisturization, integrity, hydrophobicity, texture, antimicrobial properties, emulsifier properties, mechanical properties, heat resistance or UV protection of hair, nails or skin, or a combination of two or more thereof.

[0061] In some embodiments, the composition improves skin elasticity, recovery, moisturization, integrity, texture, antimicrobial or UV protection, or a combination of two or more thereof.

[0062] In some embodiments, the composition improves the mechanical properties, color, texture, shape, moisturization, integrity, recovery, hydrophobicity or heat resistance of hair, or a combination of two or more thereof.

[0063] In some embodiments, the composition improves the mechanical properties, integrity, antimicrobial properties or UV protection of the nail, or a combination of two or more thereof.

[0064] In some embodiments, the composition further comprises urea, ammonium thioglycolate, thioanisole, benzyl alcohol, tris(hydroxymethyl)aminomethane, phosphate buffer, sodium chloride, citrate buffer, or ethanol, or a combination of two or more thereof.

[0065] In some aspects, provided herein are uses of compositions comprising fusion proteins according to embodiments of the present disclosure as agents for enhancing one or more properties of hair, nails, or skin. In some embodiments, the use of the compositions includes using the fusion proteins as agents for enhancing elasticity, moisture retention, recovery, texture, antimicrobial properties, and / or sun protection of hair, nails, and skin.

[0066] In some aspects, provided herein are fusion proteins / peptides comprising: (a) at least one keratin or keratin-related peptide block; and (b) (i) an elastin or elastin-like peptide (ELP) block, (ii) a silk or silk-like peptide (SLP) block, (iii) a collagen or collagen-like peptide block (CLP), (iv) a resilin or resilin-like peptide block (RLP), (v) an abductin or abductin peptide (ALP) block, or (vi) a combination of two or more blocks selected from (i)-(v).

[0067] In some embodiments, the fusion protein / peptide comprises: [B n1 -A m -B n -Co -B n2 ] p Formula (I), or [B n1 -C o -B n -A m -B n2 ] p Formula (II), or [B n1 -B n -C o -A m -B n2 ] p Formula (III), or [B n1 -C o -A m -B n -B n2 ] p Formula (IV), It is expressed by the formula: "A" comprises a peptide sequence comprising at least one keratin or keratin-associated peptide block, and "m" is an integer from 1 to 10; "B" comprises a peptide sequence comprising 1 to 40 amino acids, and each of "n," "n1," and "n2" is independently an integer from 0 to 25; "C" comprises a peptide sequence having the structure Dq-Er-Fs-Gt-Hu, "D" is a silk or silk-like polypeptide (SLP) block; "E" comprises a collagen or collagen-like polypeptide (CLP) block; "F" comprises an elastin or elastin-like polypeptide (ELP) block; "G" comprises a resilin or resilin-like polypeptide (RLP) block; "H" comprises an abductin or abductin-like polypeptide (ALP) block; "q" is an integer from 0 to 100, "r" is an integer from 0 to 200, "s" is an integer from 0 to 220, "t" is an integer from 0 to 80, and "u" is an integer from 0 to 70; "o" is an integer from 0 to 30, "p" is an integer from 1 to 40, At least one of "n", "n1", "n2" or "o" is not 0.

[0068] In some embodiments, each "n," "n1," and "n2" is independently an integer from 0 to 25.

[0069] In some embodiments, "o" is an integer from 1-30.

[0070] In some embodiments, each of "n," "n1," and "n2" is an integer between 0 and 25, and "o" is an integer between 1 and 30.

[0071] In some embodiments, "p" is an integer from 3 to 37. In some embodiments, "p" is an integer from 8 to 15. In some embodiments, "p" is an integer from 10 to 12.

[0072] In some embodiments, "B" comprises a peptide sequence comprising 1 to 40 amino acids. In some embodiments, "B" comprises a peptide sequence comprising 2 to 25 amino acids. In some embodiments, "B" comprises a peptide sequence comprising 4 to 25 amino acids.

[0073] In some embodiments, "B" is X n Z c and / or Z c X n and a peptide sequence having the formula "X n " is proline, leucine, isoleucine, glutamic acid, aspartic acid, glutamine, lysine, alanine, arginine or threonine, and "Z c " comprises glycine, alanine, lysine, proline, methionine, glutamine, arginine, threonine, valine, glutamic acid, aspartic acid or asparagine.

[0074] In some embodiments, at least one keratin or keratin-associated peptide block has a length of 6 to 16 amino acids, "D" comprises a polypeptide sequence comprising SEQ ID NO:24:GAGAGS, SEQ ID NO:25:GAGSGA, SEQ ID NO:26:GAGAGY, SEQ ID NO:27:GAGYGA, SEQ ID NO:28:GAGAGA, SEQ ID NO:29:GAGAGV, SEQ ID NO:30:GAGVGA, or SEQ ID NO:31:GAGAGVGY, or a combination of two or more thereof, "E" comprises a polypeptide sequence comprising SEQ ID NO:25:GAGAGS, SEQ ID NO:26:GAGAGY, SEQ ID NO:27:GAGYGA, SEQ ID NO:28:GAGAGA, SEQ ID NO:29:GAGAGV, SEQ ID NO:30:GAGVGA, or SEQ ID NO:31:GAGAGVGY, or a combination of two or more thereof, 1 Equipped with Z, "X 1 " is proline, leucine, isoleucine, glutamic acid, aspartic acid, glutamine, lysine, alanine, arginine, or threonine, Z is glycine, alanine, lysine, proline, methionine, glutamine, arginine, threonine, valine, glutamic acid, aspartic acid, or asparagine, and "F" is a nucleotide sequence selected from the group consisting of SEQ ID NO:21:VPGX 2 and / or a polypeptide sequence comprising SEQ ID NO: 13:VPAVG, 2" is arginine, histidine, lysine, aspartic acid, glutamic acid, serine, threonine, asparagine, glutamine, cysteine, glycine, alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, or tryptophan, "G" comprises a polypeptide sequence comprising SEQ ID NO:34:GGRPSDSYGAPGGGN and / or SEQ ID NO:35:GAPAQTPSSQY, and "H" comprises a polypeptide sequence comprising SEQ ID NO:36:GGFGGMGGGS, SEQ ID NO:37:MGGG, SEQ ID NO:38:FGGMG, SEQ ID NO:39:FGGMGGG, SEQ ID NO:40:GGFGGMGGG, or SEQ ID NO:41:FGGMGGGNAG, or a combination of two or more thereof. In some embodiments, the keratin or keratin-associated peptide block comprises SEQ ID NO: 8, 9, 10, 66-81, 101-103, or a fragment thereof, or a combination of two or more thereof.

[0075] In some embodiments, "A" is the peptide sequence X 3 k CX 3 5CX 3 k Equipped with "X 3 " comprises glycine, alanine, valine, isoleucine, leucine, phenylalanine, proline, serine, threonine, asparagine, glutamine, cysteine, methionine, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, histidine, lysine, or arginine, and "k" is an integer from 0 to 3.

[0076] In some embodiments, "A" comprises SEQ ID NO:8:GGVCGPSPPCITT, SEQ ID NO:9:GGVCGPSPPC, SEQ ID NO:10:CGPSPPCITT, SEQ ID NO:66:CGPSPPC, or SEQ ID NO:67:GVCGPSPPC.

[0077] In some embodiments, the fusion protein / peptide has an average molecular weight of about 1 kDa to about 144 kDa. In some embodiments, the fusion protein / peptide has an average molecular weight of about 10 kDa to about 144 kDa. In some embodiments, the fusion protein / peptide has an average molecular weight of about 35 kDa to about 130 kDa.

[0078] In some embodiments, the fusion protein / peptide is from about 1 kDa to about 5 kDa, from about 5 kDa to about 10 kDa, from about 6 kDa to about 17 kDa, from about 10 kDa to about 15 kDa, from about 14 kDa to about 30 kDa, from about 15 kDa to about 20 kDa, from about 17 kDa to about 39 kDa, from about 20 kDa to about 25 kDa, from about 25 kDa to about 30 kDa, from about 30 kDa to about 40 kDa, from about 40 kDa to about 50 kDa, from about 50 kDa to about 60 kDa, from about 60 kDa to about 70 kDa, from about 70 kDa to about 80 kDa, from about 80 kDa to about 90 kDa, from about 90 kDa to about 100 kDa, from about 100 kDa to about 150 kDa, from about 14 kDa to about 30 kDa, from about 15 kDa to about 20 kDa, from about 17 kDa to about 39 kDa, from about 20 kDa to about 25 kDa, from about 25 kDa to about 30 kDa, from about 30 kDa to about 40 kDa, from about 40 kDa to about 50 kDa, from about 50 kDa to about 60 kDa, from about 60 kDa to about 70 kDa, from about 70 kDa to about 80 kDa, from about 80 kDa to about 90 kDa, from about 90 kDa to about 100 kDa, from about 10 The average molecular weight ranges are: about 35 kDa to about 35 kDa, about 35 kDa to about 40 kDa, about 39 kDa to about 54 kDa, about 39 kDa to about 80 kDa, about 40 kDa to about 45 kDa, about 45 kDa to about 50 kDa, about 50 kDa to about 55 kDa, about 55 kDa to about 60 kDa, about 60 kDa to about 100 kDa, and about 80 kDa to about 144 kDa.

[0079] In some embodiments, the fusion protein has a secondary structure. In some embodiments, the secondary structure is an alpha (α) helix, a triple helix, or a triple helix. 10 It comprises a helix, a pi (π) helix, a beta (β) turn, a β sheet, a coil, or a combination thereof.

[0080] In some embodiments, the silk or silk-like peptide (SLP) block comprises an α-helical, 3 10 Forms a helix, a π-helix, a beta (β) turn, a β-sheet, a coil, or a combination thereof. In some embodiments, the silk or silk-like peptide (SLP) block forms a β-sheet rich structure.

[0081] In some embodiments, the elastin or elastin-like peptide (ELP) block comprises an alpha helix, a triad 10Forms a helix, a π-helix, a beta (β) turn, a β-sheet, a coil, or a combination thereof. In some embodiments, the elastin or elastin-like peptide (ELP) block forms a β-turn-rich structure.

[0082] In some embodiments, the linker block is an α-helix, 3 10 In some embodiments, the linker blocks form a helix, a π-helix, a beta (β) turn, a β-sheet, a coil, or a combination thereof. In some embodiments, the linker blocks form an α-helix. In some embodiments, the linker blocks form a coil. In some embodiments, the linker blocks form a coil.

[0083] In some embodiments, the elastin-like peptide (ELP) block is temperature responsive. In some embodiments, the ELP block may respond to changes in pH. In some embodiments, the ELP block may respond over a range of temperatures. In some embodiments, the ELP block may respond to salt.

[0084] In some aspects, the description provides for the use of a fusion protein, fusion protein / peptide or composition according to embodiments of the present disclosure as a cosmetic, such as a hair, nail or skin cosmetic.

[0085] In some embodiments, the cosmetic product is suitable for hair care or hair treatment.

[0086] In some aspects, the present specification provides methods for obtaining the fusion proteins of the present disclosure, comprising the steps of inoculating a liquid culture medium with microorganisms containing an expression gene for the desired fusion protein, allowing the microorganisms to grow for a specific culture period and inducing expression under the influence of a promoter, and recovering the desired fusion protein from the resulting microorganisms by removing endogenous contaminants.

[0087] In some embodiments, the microorganism is Escherichia coli. In some embodiments, the microorganism is Komagataella phaffii.

[0088] In some embodiments, the fusion protein is purified from endogenous contaminants.

[0089] Another aspect relates to a composition comprising the fusion protein / peptide of the present disclosure for improving the elasticity, recovery, moisturization, integrity, hydrophobicity, texture, antimicrobial properties, emulsifier properties, mechanical properties, heat resistance or UV protection of hair, nails or skin, or a combination of two or more thereof.

[0090] In some embodiments, the composition improves skin elasticity, recovery, moisturization, integrity, texture, antimicrobial or UV protection, or a combination of two or more thereof.

[0091] In some embodiments, the amount of fusion protein / peptide ranges from 0.0001% to 20% (w / w), preferably the amount of fusion protein / peptide ranges from 0.001% to 5% (w / w), 0.1% to about 1% (w / w).

[0092] In some embodiments, the composition improves the mechanical properties, color, texture, shape, moisturization, integrity, recovery, hydrophobicity or heat resistance of hair, or a combination of two or more thereof.

[0093] In some embodiments, the composition improves the mechanical properties, integrity, antimicrobial properties or UV protection of the nail, or a combination of two or more thereof.

[0094] In some embodiments, the at least one excipient suitable for skin application is selected from a surfactant, an emulsifier, a preservative, a thickener, an organic polymer, a humectant, a silicone, an oil, a fragrance, a vitamin, a buffering agent, an antimicrobial agent, an antibacterial agent, a disinfectant, a chelating agent, or a mixture thereof.

[0095] In some embodiments, the composition may comprise at least one excipient suitable for use on the skin.

[0096] In some embodiments, the at least one excipient suitable for use on the skin is selected from ethanol, benzyl alcohol, a diol molecule, urea, ammonium thioglycolate, thioanisole, tris(hydroxymethyl)aminomethane, phosphate buffer, sodium chloride, citrate buffer, or ethanol, or a combination of two or more thereof.

[0097] In some embodiments, the diol molecule is selected from ethylene glycol, propylene glycol, butylene glycol, butylene diol, or mixtures thereof.

[0098] In some embodiments, the composition may comprise 0-30% (w / v), preferably 10-20% (w / v), ethanol, 0-3% (w / v), preferably 0.5-2% (w / v), benzyl alcohol, and 0-20% (w / v), preferably 10-15% (w / v), diol molecules.

[0099] In some embodiments, the composition may further comprise a propellant, a fragrance, an oil, or a mixture thereof.

[0100] In some embodiments, the composition may be used to treat hair.

[0101] The fusion proteins / peptides or compositions of the present disclosure may be used as cosmetics, i.e., hair, nail or skin cosmetics.

[0102] In some embodiments, the cosmetic product is suitable for hair care or hair treatment.

[0103] In another aspect, the present disclosure relates to shampoos, lotions, serums, creams, conditioners, foams, elixirs, oils, aerosols, or masks comprising the compositions / fusion proteins / peptides of the present disclosure.

[0104] In another aspect, the present disclosure relates to the use of a composition comprising a fusion protein of the present disclosure as an enhancer of one or more properties of hair, nails, or skin, or as an agent for enhancing the elasticity, moisture retention, resilience, texture, antimicrobial properties, and / or sun protection of hair, nails, and skin.

[0105] Another aspect relates to a method for obtaining the fusion protein / peptide of the present disclosure, comprising the steps of inoculating a liquid culture medium with a microorganism containing an expression gene for the desired fusion protein, growing the microorganism for a specific cultivation period and inducing expression under the influence of a promoter, and removing endogenous contaminants from the obtained microorganism to recover the desired fusion protein / peptide.

[0106] In one embodiment, the microorganism is Escherichia coli.

[0107] In one embodiment, the fusion protein / peptide is purified from endogenous contaminants.

[0108] Another aspect relates to a method for obtaining the fusion proteins / peptides of the present disclosure by solid phase synthesis, comprising the steps of initiating solid phase synthesis by combining a keratin or keratin-related peptide block with at least one polypeptide block selected from the list consisting of (i) an elastin or elastin-like polypeptide (ELP) block, (ii) a silk or silk-like polypeptide (SLP) block, (iii) a collagen or collagen-like polypeptide (CLP) block, (iv) a resilin or resilin-like polypeptide (RLP) block, (v) an abductin or abductin-like polypeptide (ALP) block, or (vi) a combination of two or more blocks selected from (i) to (v), and performing multiple solid phase reactions to obtain the fusion protein / peptide.

[0109] The following figures show examples to illustrate the present disclosure and are not intended to limit the scope of the present disclosure. [Brief explanation of the drawings]

[0110] [Figure 1] 1 shows a linear vector map of an expression vector comprising a promoter, a target fusion protein sequence, a linker sequence, a C-terminal affinity tag sequence, and an AmpR gene and corresponding promoter according to an embodiment of the present disclosure. [Figure 2] 1 shows a linear vector map of an expression vector comprising a promoter, an N-terminal affinity tag sequence, a linker sequence, a target fusion protein, and an AmpR gene and corresponding promoter according to an embodiment of the present disclosure. [Figure 3] Fluorescence microscopy of peptides labeled with FITC (a fluorescent probe) and applied to hair fibers shows their penetration / location. Most peptides, especially peptides SEQ ID NO:2, SEQ ID NO:45, SEQ ID NO:46 and SEQ ID NO:47, were able to penetrate from the cuticle to the cortex. DETAILED DESCRIPTION OF THE INVENTION

[0111] The present disclosure relates to fusion proteins / peptides.

[0112] One aspect of the present disclosure relates to fusion proteins / peptides comprising effective amounts of repeat sequences of various proteins that can be used to improve the properties of hair, skin, and nails.

[0113] Another aspect of the present disclosure relates to compositions comprising the fusion proteins / peptides of the present disclosure.

[0114] In some embodiments, the fusion proteins / peptides may be used in hair care compositions, skin care compositions, nail care compositions, cosmetic compositions, or pharmaceutical compositions.

[0115] In some embodiments, fusion proteins / peptides and compositions comprise effective amounts of repeating sequences of different proteins / peptides, which has the advantage that the fusion proteins / peptides can provide compositions with desired properties, which can include elasticity and stimuli-responsive behavior.

[0116] Keratin and keratin-associated proteins are the main components of human hair, skin, and nails. Keratin and keratin-associated proteins typically have a high sulfur content due to the presence of cysteine ​​amino acids. The presence of sulfur is essential for the structure of hair, skin, and nails because it allows for the formation of disulfide bonds between peptide chains. The presence of disulfide bonds (i.e., cross-links) is a major contributor to the structure and texture of hair, skin, and nails. Damage to the disulfide bridges (i.e., bonds) between keratins can alter the appearance and texture of hair, skin, and nails. Maintaining the integrity of the disulfide bonds between keratins can help keep hair, skin, and nails healthy and prevent cracking and breakage.

[0117] (fusion protein) In some embodiments, the fusion protein is a keratin fusion protein comprising a keratin or keratin-associated peptide block derived from a keratin protein fraction. The keratin and keratin-associated fusion proteins described herein can bind with high affinity to hair, skin, and nails and repair damage to keratin fibers through the formation of new disulfide bonds. The keratin or keratin-associated peptide block of the fusion protein can have a sequence containing one or more cysteine ​​amino acids that naturally form disulfide bonds with other free cysteine ​​amino acids in hair, skin, and nails. The formation of new disulfide bonds can help restore the mechanical properties of hair, skin, and nails, including elasticity, durability, and appearance.

[0118] In some embodiments, the fusion proteins disclosed herein comprise amino acid sequences (the term "peptide" may be used interchangeably to refer to amino acid sequences) that can be joined by covalent bonds or any other type of bond that results in a structure that maintains the properties of the fusion proteins of the present disclosure. The bond may be selected from, but is not limited to, hydrogen bonding, ion pairing, hydrophobic association, or inclusion complex formation.

[0119] In some embodiments, the keratin or keratin-associated fusion protein / peptide comprises at least one keratin or keratin-associated peptide block.

[0120] In some embodiments, the keratin, keratin peptide, or keratin-associated peptide comprises a wild-type keratin peptide, a functional fragment of a wild-type keratin peptide, or a peptide having at least 90% identity to a wild-type keratin peptide or a functional fragment of a wild-type keratin peptide. The wild-type keratin peptide can be keratin 85 having SEQ ID NO: 48. The wild-type peptide keratin peptide can have SEQ ID NO: 8, 9, 10, 66, or 67. In some embodiments, the keratin, keratin peptide, or keratin-associated peptide forms one or more disulfide bonds with hair. In some embodiments, the keratin, keratin peptide, or keratin-associated peptide forms one or more disulfide bonds with skin. In some embodiments, the keratin, keratin peptide, or keratin-associated peptide forms one or more disulfide bonds with nails.

[0121] As a non-limiting example, a functional fragment of a wild-type keratin peptide comprises at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 consecutive amino acids of the wild-type keratin peptide. As a non-limiting example, a functional fragment of a wild-type keratin peptide comprises at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 amino acids that are at least 90% identical to the wild-type keratin peptide. As a non-limiting example, a functional fragment of a wild-type keratin peptide comprises at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 amino acids, wherein the at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 amino acids comprise one or more cysteine ​​amino acids. For example, the one or more cysteine ​​amino acids are 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or 27 cysteine ​​amino acids. As a non-limiting example, a functional fragment of a wild-type keratin peptide comprises at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 amino acids, wherein at least 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 30%, 40%, or 50% of the at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 amino acids are cysteine ​​amino acids. A non-limiting example of a keratin or keratin peptide is shown in SEQ ID NO:48.

[0122] In some embodiments, keratin-associated peptides (sometimes referred to as keratin-associated proteins) include peptides that form disulfide bonds with wild-type keratin. In some embodiments, keratin-associated peptides comprise at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 amino acids, and at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 amino acids are at least 90% identical to the wild-type keratin peptide sequence. The wild-type keratin peptide may be keratin 85, having SEQ ID NO: 48. The wild-type keratin peptide may comprise SEQ ID NO: 8, 9, 10, 66, or 67.

[0123] In some embodiments, the keratin or keratin-associated peptide component of the fusion protein has from about 6 to about 16 amino acids.

[0124] In some embodiments, the keratin or keratin-associated peptide component of the fusion protein has about 6 amino acids.

[0125] In some embodiments, the keratin or keratin-associated peptide component of the fusion protein has about 7 amino acids.

[0126] In some embodiments, the keratin or keratin-associated peptide component of the fusion protein has about 8 amino acids.

[0127] In some embodiments, the keratin or keratin-associated peptide component of the fusion protein has about 9 amino acids.

[0128] In some embodiments, the keratin or keratin-associated peptide component of the fusion protein has about 10 amino acid residues.

[0129] In some embodiments, the keratin or keratin-associated peptide component of the fusion protein has about 11 amino acid residues.

[0130] In some embodiments, the keratin or keratin-associated peptide component of the fusion protein has about 12 amino acids.

[0131] In some embodiments, the keratin or keratin-associated peptide component of the fusion protein has about 13 amino acids.

[0132] In some embodiments, the keratin or keratin-associated peptide component of the fusion protein has about 14 amino acids.

[0133] In some embodiments, the keratin or keratin-associated peptide component of the fusion protein has about 15 amino acid residues.

[0134] In some embodiments, the keratin or keratin-associated peptide component of the fusion protein has about 16 amino acids.

[0135] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein has a high cysteine ​​content to enhance certain properties of the keratin or keratin-associated peptide block, such as enhanced intermolecular cross-linking, and the total cysteine ​​content ranges from 10% to 50% of the total number of amino acids in the keratin or keratin-associated peptide block.

[0136] In some embodiments, the total cysteine ​​content of the keratin or keratin-associated peptide of the fusion protein / peptide is greater than 15% of the total number of amino acids in the keratin or keratin-associated peptide block.

[0137] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein / peptide comprises two cysteine ​​residues.

[0138] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein comprises three cysteine ​​residues.

[0139] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein comprises four cysteine ​​residues.

[0140] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein comprises five cysteine ​​residues.

[0141] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein / peptide comprises the peptide sequence (X 3 ) k C(X 3 )5C(X 3 ) k Equipped with.

[0142] In some embodiments, each "X 3 are independently selected from glycine, alanine, valine, isoleucine, leucine, phenylalanine, proline, serine, threonine, asparagine, glutamine, cysteine, methionine, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, histidine, lysine, or arginine.

[0143] In some embodiments, each "k" is independently an integer from 0 to 3. When k is 2, (X 3 ) k The first X 3 and the second X 3 and each X 3 are independently glycine, alanine, valine, isoleucine, leucine, phenylalanine, proline, serine, threonine, asparagine, glutamine, cysteine, methionine, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, histidine, lysine, or arginine. When k is 3, (X 3 ) k The first X 3 , the second X 3 and a third X 3 and each X 3are independently glycine, alanine, valine, isoleucine, leucine, phenylalanine, proline, serine, threonine, asparagine, glutamine, cysteine, methionine, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, histidine, lysine, or arginine. (X 3 )5 is the first X 3 , the second X 3 , the third X 3 , 4th X 3 and the fifth X 3 and each X 3 are independently glycine, alanine, valine, isoleucine, leucine, phenylalanine, proline, serine, threonine, asparagine, glutamine, cysteine, methionine, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, histidine, lysine, or arginine.

[0144] In some embodiments, the keratin or keratin-associated peptide component of the fusion protein comprises the sequence of GGVCGPSPPCITT (SEQ ID NO:8), GGVCGPSPPC (SEQ ID NO:9), CGPSPPCITT (SEQ ID NO:10), CGPSPPC (SEQ ID NO:66), GVCGPSPPC (SEQ ID NO:67), or SEQ ID NOs:70-81, or a combination of two or more thereof.

[0145] In some embodiments, the keratin or keratin-associated peptide sequence is GGVCGPSPPCITT (SEQ ID NO:8). In some embodiments, the keratin or keratin-associated peptide sequence is GGVCGPSPPC (SEQ ID NO:9). In some embodiments, the keratin or keratin-associated peptide sequence is CGPSPPCITT (SEQ ID NO:10).

[0146] In some embodiments, the keratin or keratin-related fusion protein further comprises an elastin or elastin-like polypeptide (ELP) block, a silk or silk-like polypeptide (SLP) block, a collagen or collagen-like polypeptide (CLP) block, a resilin or resilin-like polypeptide (RLP) block, an abductin or abductin-like polypeptide (ALP) block, or a combination of two or more thereof, such as a silk-elastin-like polypeptide (SELP) block.

[0147] In some embodiments, the keratin or keratin-associated fusion proteins disclosed herein further comprise an elastin or elastin-like polypeptide (ELP) block. In some embodiments, elastin-like polypeptide (ELP) refers to a peptide, including a synthetic peptide, comprising an amino acid sequence having a similar secondary structure, tertiary structure, or amino acid sequence to a wild-type elastin protein (e.g., SEQ ID NO: 55) or a fragment of the wild-type elastin protein, or a combination of two or more thereof. In some embodiments, the fragment comprises a sequence having at least 4, 5, 6, 7, 8, 9, or 10 amino acids that are 100% identical to the sequence of the wild-type elastin protein. Non-limiting examples of elastins and ELPs include SEQ ID NOs: 11-23, 49-55, and 82-91.

[0148] The ELP may comprise a polymeric or oligomeric repeating peptide sequence of elastin protein. In one exemplary embodiment, the elastin or ELP comprises SEQ ID NO:49 ((VPGXG)n, where n is 1-10 and each X is independently any amino acid), or a sequence having up to one amino acid substitution compared to SEQ ID NO:49. In some embodiments, each X is independently valine or leucine. In some embodiments, n is 5. In some embodiments, n is 6.

[0149] In one exemplary embodiment, the elastin or ELP comprises SEQ ID NO:50 (VPGVG), or a sequence having up to one amino acid substitution compared to SEQ ID NO:50. The elastin or ELP can comprise 1-10 sequences having SEQ ID NO:50, or 1-10 sequences each independently having up to one amino acid substitution compared to SEQ ID NO:50. The 1-10 sequences can be linked, for example, VPGVGVPGVGVPGVGVPGVGVPGVG (SEQ ID NO:23), or separated by another ELP, such as in one or more amino acids, for example, VPGVGVPGVGVPGLGVPGVGVPGVG (SEQ ID NO:54).

[0150] In one exemplary embodiment, the elastin or ELP comprises SEQ ID NO:51 (VPGLG), or a sequence having no more than one amino acid substitution compared to SEQ ID NO:51. The elastin or ELP may comprise 1 to 10 sequences having SEQ ID NO:51, or 1 to 10 sequences each independently having no more than one amino acid substitution compared to SEQ ID NO:51. The 1 to 10 sequences may be linked or separated by one or more amino acids.

[0151] In one exemplary embodiment, the elastin or ELP comprises SEQ ID NO:52 (VPGVGVPGL), or a sequence having up to one amino acid substitution compared to SEQ ID NO:52. The elastin or ELP may comprise 1 to 10 sequences having SEQ ID NO:52, or 1 to 10 sequences each independently having up to one amino acid substitution compared to SEQ ID NO:52. The 1 to 10 sequences may be linked or separated by one or more amino acids.

[0152] In one exemplary embodiment, the elastin or ELP comprises SEQ ID NO: 53 (VPGVGVPGVGVPGLGVPGVGVPGVGR), or a sequence having up to one amino acid substitution compared to SEQ ID NO: 53. The elastin or ELP may comprise 1 to 10 sequences having SEQ ID NO: 53, or 1 to 10 sequences each independently having up to one amino acid substitution compared to SEQ ID NO: 53. The 1 to 10 sequences may be linked or separated by one or more amino acids.

[0153] In one exemplary embodiment, the elastin or ELP comprises SEQ ID NO:23 (VPGVGVPGVGVPGVGVPGVGVPGVGVPGVG), or a sequence having up to one amino acid substitution compared to SEQ ID NO:23. The elastin or ELP may comprise 1 to 10 sequences having SEQ ID NO:23, or 1 to 10 sequences each independently having up to one amino acid substitution compared to SEQ ID NO:23. The 1 to 10 sequences may be linked or separated by one or more amino acids.

[0154] In one exemplary embodiment, the elastin or ELP comprises SEQ ID NO:54 (VPGVGVPGVGVPGLGVPGVGVPGVG), or a sequence having up to one amino acid substitution compared to SEQ ID NO:54. The elastin or ELP may comprise 1 to 10 sequences having SEQ ID NO:54, or 1 to 10 sequences each independently having up to one amino acid substitution compared to SEQ ID NO:54. The 1 to 10 sequences may be linked or separated by one or more amino acids.

[0155] In one exemplary embodiment, the elastin or ELP comprises SEQ ID NO:22 (GVGVPGVGVPGLGVPGVGVPGVG), or a sequence having up to one amino acid substitution compared to SEQ ID NO:22. The elastin or ELP may comprise one to ten sequences having SEQ ID NO:22, or one to ten sequences each independently having up to one amino acid substitution compared to SEQ ID NO:22. The one to ten sequences may be linked or separated by one or more amino acids.

[0156] In some embodiments, the elastin or ELP comprises polymeric or oligomeric repeating peptide sequences of elastin peptides, hi some embodiments, each polymeric or oligomeric repeating peptide sequence comprises or has 4 amino acid residues.

[0157] In some embodiments, each polymeric or oligomeric repeating peptide sequence comprises or has 5 amino acid residues. Non-limiting examples include SEQ ID NOs: 49, 50, or 51.

[0158] In some embodiments, each polymeric or oligomeric repeating peptide sequence comprises or has 6 amino acid residues.

[0159] In some embodiments, each polymeric or oligomeric repeat peptide sequence comprises or has 7 amino acid residues.

[0160] In some embodiments, each polymeric or oligomeric repeat peptide sequence comprises or has 8 amino acid residues.

[0161] Non-limiting examples of repeating peptide sequences include polytetrapeptides, polypentapeptides, polyhexapeptides, polyheptapeptides, polyoctapeptides, and polynonapeptides, including VPGG (SEQ ID NO:11), IPGG (SEQ ID NO:12), VPAVG (SEQ ID NO:13), AVGVP (SEQ ID NO:14), IPGVG (SEQ ID NO:15), LPGVG (SEQ ID NO:16), VAPGVG (SEQ ID NO:17), GVGVPGVG (SEQ ID NO:18), VPGFGVGAG (SEQ ID NO:19), VPGVGVPGG (SEQ ID NO:20), VPGX 2 G (SEQ ID NO:21), SEQ ID NO:49 ((VPGXG)n, where n is 1-10 and each X is independently any amino acid), SEQ ID NO:50 (VPGVG), SEQ ID NO:51 (VPGLG), and SEQ ID NO:52 (VPGVGVPGL), and combinations of two or more thereof.

[0162] In some embodiments, "X 2 " is a natural or unnatural amino acid.

[0163] In some embodiments, the elastin or ELP is a polymeric or oligomeric repeat of the pentapeptide VPAVG (SEQ ID NO:13).

[0164] In some embodiments, elastin or ELP is the pentapeptide VPGX 2 G is a polymer or oligomer of repeating "X 2 " is an amino acid selected from arginine, histidine, lysine, aspartic acid, glutamic acid, serine, threonine, asparagine, glutamine, cysteine, glycine, alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, or tryptophan. In some embodiments, "X 2 " may be leucine. In some embodiments, "X 2" is asparagine.

[0165] In some embodiments, "X 2 " is Balin.

[0166] In some embodiments, elastin or ELP is defined by the peptide sequence GVGVPGVGVPGLGVPGVGVPGVG (SEQ ID NO:22).

[0167] In other embodiments, elastin or ELP is defined by the sequence VPGVGVPGVGVPGVGVPGVGVPGVGVPGVG (SEQ ID NO:23).

[0168] In some embodiments, the elastin or elastin-like polypeptide (ELP) component forms a beta-sheet rich domain.

[0169] In some embodiments, the elastin or ELP component comprises amorphous domains. In some embodiments, the elastin or ELP component comprises crystalline domains. In some embodiments, the elastin or ELP component comprises both crystalline and amorphous domains.

[0170] In some embodiments, the elastin or ELP component is stimulus-responsive. In some embodiments, the ELP component is temperature-responsive. In some embodiments, the ELP component may respond to changes in pH. In some embodiments, the ELP component may respond over a range of temperatures. In some embodiments, the ELP component may respond to salt.

[0171] In some embodiments, the keratin or keratin-related fusion proteins / peptides disclosed herein comprise a silk or silk-like polypeptide (SLP) component. In some embodiments, a silk-like polypeptide (SLP) refers to a peptide, including synthetic peptides, that has a secondary structure, tertiary structure, or amino acid sequence similar to that of a wild-type silk protein (e.g., SEQ ID NO: 56) or a fragment of a wild-type silk protein, or a combination of two or more thereof. In some embodiments, the fragment comprises a sequence having at least 4, 5, 6, 7, 8, 9, or 10 amino acids that are 100% identical to the sequence of the wild-type silk protein. The wild-type silk protein can be fibroin. Non-limiting examples of silks and SLPs include SEQ ID NOs: 24-32, 56, and 92.

[0172] The SLP may comprise a polymeric or oligomeric repeating peptide sequence of a silk protein. In one exemplary embodiment, the silk or SLP comprises SEQ ID NO:24 (GAGAGS), or a sequence having up to one amino acid substitution compared to SEQ ID NO:24. In one exemplary embodiment, the silk or SLP comprises SEQ ID NO:24, or a sequence having up to one amino acid substitution compared to SEQ ID NO:24. The silk or SLP may comprise 1 to 10 sequences having SEQ ID NO:24, or 1 to 10 sequences each independently having up to one amino acid substitution compared to SEQ ID NO:24. The 1 to 10 sequences may be linked or separated by one or more amino acids.

[0173] In some embodiments, the silk or SLP comprises polymeric or oligomeric repeating peptide sequences of silk peptides, each polymeric or oligomeric repeating peptide sequence comprising or having 6 amino acid residues.

[0174] In some embodiments, the silk or SLP comprises polymeric or oligomeric repeating peptide sequences of silk peptides, each polymeric or oligomeric repeating peptide sequence comprising or having 7 amino acid residues.

[0175] In some embodiments, the silk or SLP comprises polymeric or oligomeric repeating peptide sequences of silk peptides, each polymeric or oligomeric repeating peptide sequence comprising or having 8 amino acid residues.

[0176] Non-limiting examples of repeat peptide sequences include polytetrapeptides, polypentapeptides, polyhexapeptides, polyheptapeptides, polyoctapeptides, and polynonapeptides, including GAGAGS (SEQ ID NO:24), GAGSGA (SEQ ID NO:25), GAGAGY (SEQ ID NO:26), GAGYGA (SEQ ID NO:27), GAGAGA (SEQ ID NO:28), GAGAGV (SEQ ID NO:29), GAGVGA (SEQ ID NO:30), GAGAGVGY (SEQ ID NO:31), and combinations of two or more thereof.

[0177] In some embodiments, silks or SLPs are polymeric or oligomeric repeats of the pentapeptide GAGAGS (SEQ ID NO:24).

[0178] In some embodiments, the silk or SLP has the peptide sequence GAGAGSGAGAGSGAGAGSGAGAGSGAGAGS (SEQ ID NO:32).

[0179] In some embodiments, the silk or SLP has the peptide sequence GAGX 1 GX 2 (SEQ ID NO:92), and X 1 and X 2 are independently any amino acid, and optionally X 1 is A, S, Y or V, and optionally X 2is S, A, Y or V.

[0180] In some embodiments, the silk or SLP is the peptide sequence GYGAGAGSGAASGAGAGSGAGAGSGAGAGSGAGAGSGAGAGSGAGAGSGAGAGYGAGAGVGYGAGAGSGAASGAGAGSGAGAGSGAGAGSGAGAGS (SEQ ID NO:56).

[0181] In some embodiments, the silk or silk-like (SLP) blocks form a beta-sheet rich domain.

[0182] In some embodiments, the silk or SLP component blocks form crystalline domains. In some embodiments, the silk or SLP blocks comprise amorphous domains. In some embodiments, the SLP blocks comprise both crystalline and amorphous domains.

[0183] In some embodiments, the keratin or keratin-related fusion proteins disclosed herein comprise a collagen or collagen-like polypeptide (CLP) block. In some embodiments, collagen-like polypeptide (CLP) refers to a peptide, including synthetic peptides, that has a similar secondary structure, tertiary structure, or amino acid sequence, or a combination of two or more thereof, to a wild-type collagen protein (e.g., SEQ ID NO: 59) or a fragment of a wild-type collagen protein. In some embodiments, the fragment comprises a sequence having at least 3, 4, 5, 6, 7, 8, 9, or 10 amino acids that are 100% identical to the sequence of the wild-type collagen protein. Non-limiting examples of collagens and CLPs include SEQ ID NOs: 33, 57-59, 93-95, and 106, and fragments having the amino acids "GTP" and / or "GLQ."

[0184] A CLP may comprise a polymeric or oligomeric repeating peptide sequence of a collagen peptide. In an exemplary embodiment, the collagen or CLP comprises a GTP sequence. The collagen or CLP may comprise 1 to 10 sequences comprising a GTP sequence. The 1 to 10 sequences may be linked and separated by one or more amino acids. The 1 to 10 sequences may be, for example, two or three. For example, the collagen or CLP may comprise GPTGPT (SEQ ID NO: 57), GLQGLQ (SEQ ID NO: 58), GPAGPA (SEQ ID NO: 95), or GPTGPTGPAGPRGLQGLQGLQGERGEQGPT (SEQ ID NO: 33). In an exemplary embodiment, the collagen or CLP comprises a GLQ sequence. The collagen or CLP may comprise 1 to 10 sequences comprising a GLQ sequence. The 1 to 10 sequences may be, for example, two or three. The 1 to 10 sequences may be linked and separated by one or more amino acids. For example, collagen or CLP may comprise GLQGLQ (SEQ ID NO:58) or GPTGPTGPAGPRGLQGLQGLQGERGEQGPT (SEQ ID NO:33). In an exemplary embodiment, collagen or CLP comprises a GPT sequence. Collagen or CLP may comprise 1 to 10 sequences comprising a GPT sequence. The 1 to 10 sequences may be, for example, two or three. The 1 to 10 sequences may be linked and separated by one or more amino acids. For example, collagen or CLP may comprise GPTGPT (SEQ ID NO:57) or GPTGPTGPAGPRGLQGLQGLQGERGEQGPT (SEQ ID NO:33). Collagen or CLP may comprise 1 to 10 sequences comprising a GXP sequence (X is any amino acid). The 1 to 10 sequences may be, for example, two or three. The 1 to 10 sequences may be linked and separated by one or more amino acids. For example, collagen or CLP may comprise GXPGXP (SEQ ID NO:93). Collagen or CLP may comprise 1 to 10 sequences comprising a GZP sequence, where Z is 4-hydroxyproline. The 1 to 10 sequences may be, for example, 2 or 3.The 1 to 10 sequences can be linked and separated by one or more amino acids, for example, collagen or CLP can comprise GLQGLQ (SEQ ID NO:94).

[0185] In some embodiments, each polymeric or oligomeric repeating peptide sequence of the collagen or collagen-like polypeptide (CLP) block comprises or has three amino acid residues. Non-limiting examples of the three amino acid residues include GTP, GLQ, GXP (where X is any amino acid), GZP (where Z is 4-hydroxyproline), and GX 1 It may contain H.

[0186] In some embodiments, the repeating peptide sequences include polytetrapeptides, polypentapeptides, polyhexapeptides, polyheptapeptides, polyoctapeptides, and polynonapeptides, 1 Including but not limited to H.

[0187] In some embodiments, "X 1 " comprises proline, leucine, isoleucine, glutamic acid, aspartic acid, glutamine, lysine, alanine, arginine and threonine, and "H" comprises an amino acid selected from glycine, alanine, lysine, proline, methionine, glutamine, arginine, threonine, valine, glutamic acid, aspartic acid or asparagine.

[0188] In some embodiments, the collagen or CLP is defined by the peptide sequence GPTGPTGPAGPRGLQGLQGLQGERGEQGPT (SEQ ID NO:33).

[0189] In some embodiments, the collagen or CLP is defined by the peptide sequence SEQ ID NO:59.

[0190] In some embodiments, the collagen or CLP forms an alpha helix (α helix). In some embodiments, multiple collagens or CLPs form a triple helix. In some embodiments, the collagen or CLP has a 3.6 13 Forms a helix. In some embodiments, the collagen or CLP forms a polyproline type II helix. In some embodiments, the collagen or CLP forms at least one helix loop.

[0191] In some embodiments, the keratin or keratin-associated fusion proteins disclosed herein comprise a resilin or resilin-like polypeptide (RLP) block. In some embodiments, resilin-like polypeptide (RLP) refers to a peptide, including a synthetic peptide, that has a similar secondary structure, tertiary structure, or amino acid sequence, or a combination of two or more thereof, to a wild-type resilin protein (e.g., SEQ ID NO: 60) or a fragment of a wild-type resilin protein. In some embodiments, the fragment comprises a sequence having 4, 5, 6, 7, 8, 9, or 10 amino acids that are 100% identical to the sequence of the wild-type silk protein. Non-limiting examples of resilin and RLP include SEQ ID NOs: 34-35, 60, 96, and 107.

[0192] The RLP can comprise a polymeric or oligomeric repeating peptide sequence of a resilin peptide. In exemplary embodiments, the polymeric or oligomeric repeating peptide sequence is a fragment of a wild-type resilin peptide having about 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids.

[0193] In some embodiments, each polymeric or oligomeric repeat peptide sequence of a resilin or resilin-like polypeptide (RLP) block comprises or has 11 amino acid residues. In some embodiments, each polymeric or oligomeric repeat peptide sequence of a resilin or resilin-like polypeptide (RLP) block comprises or has 12 amino acid residues. In some embodiments, each polymeric or oligomeric repeat peptide sequence of a resilin or resilin-like polypeptide (RLP) block comprises or has 13 amino acid residues. In some embodiments, each polymeric or oligomeric repeat peptide sequence of a resilin or resilin-like polypeptide (RLP) block comprises or has 14 amino acid residues. In some embodiments, each polymeric or oligomeric repeat peptide sequence of a resilin or resilin-like polypeptide (RLP) block comprises or has 15 amino acid residues. In some embodiments, the repeating peptide sequence comprises a polypentapeptide, a polyhexapeptide, a polyheptapeptide, a polyoctapeptide, and a polynonapeptide, including, but not limited to, GGRPSDSYGAPGGGN (SEQ ID NO:34), GAPAQTPSSQY (SEQ ID NO:35), AQTPSSQYGAP (SEQ ID NO:96), and combinations thereof. In some embodiments, resilin or RLP is defined by the peptide sequence GGRPSDSYGAPGGGN (SEQ ID NO:34). In some embodiments, resilin or RLP is defined by the peptide sequence SEQ ID NO:60. In some embodiments, resilin or RLP is defined by the peptide sequence SEQ ID NO:107. In some embodiments, resilin or RLP is defined by the peptide sequence GAPAQTPSSQY (SEQ ID NO:35). In some embodiments, resilin or RLP is defined by the peptide sequence AQTPSSQYGAP (SEQ ID NO:96).

[0194] In some embodiments, the keratin or keratin-associated fusion proteins disclosed herein comprise an abductin or abductin-like polypeptide (ALP) block. In some embodiments, an abductin-like polypeptide (ALP) refers to a peptide, including a synthetic peptide, comprising an amino acid sequence having a similar secondary structure, tertiary structure, or amino acid sequence to a wild-type abductin protein (e.g., SEQ ID NO: 61) or a fragment of the wild-type abductin protein, or a combination of two or more thereof. In some embodiments, the fragment comprises a sequence having 4, 5, 6, 7, 8, 9, or 10 amino acids that are 100% identical to the sequence of the wild-type abductin protein. Non-limiting examples of abductins and ALPs include SEQ ID NOs: 36-42 and 61. An ALP may comprise a polymeric or oligomeric repeating peptide sequence of an abductin protein.

[0195] In some embodiments, each polymeric or oligomeric repeat peptide sequence comprises or has 4 amino acid residues of an abductin protein. In some embodiments, each polymeric or oligomeric repeat peptide sequence comprises or has 5 amino acid residues of an abductin protein. In some embodiments, each polymeric or oligomeric repeat peptide sequence comprises or has 6 amino acid residues of an abductin protein.

[0196] In some embodiments, each polymeric or oligomeric repeat peptide sequence comprises or has 7 amino acid residues of an abductin protein. In some embodiments, each polymeric or oligomeric repeat peptide sequence comprises or has 8 amino acid residues of an abductin protein. In some embodiments, each polymeric or oligomeric repeat peptide sequence comprises or has 9 amino acid residues of an abductin protein. In some embodiments, each polymeric or oligomeric repeat peptide sequence comprises or has 10 amino acid residues of an abductin protein. In some embodiments, the repeating peptide sequence comprises a polytetrapeptide, a polypentapeptide, a polyhexapeptide, a polyheptapeptide, a polyoctapeptide, and a polynonapeptide, including, but not limited to, GGFGGMGGGS (SEQ ID NO:36), MGGG (SEQ ID NO:37), FGGMG (SEQ ID NO:38), FGGMGGG (SEQ ID NO:39), GGFGGMGGG (SEQ ID NO:40), FGGMGGGNAG (SEQ ID NO:41), and combinations thereof. In some embodiments, abductin or ALP is defined by the peptide sequence GGFGGMGGGSGGFGGMGGGSGGFGGMGGGS (SEQ ID NO:42). In some embodiments, abductin or ALP is defined by the peptide sequence MNAYICLAACLIAAVSAAGYGGGAGSMGGTGGMGGGMNAGGFGGMGGMGGGKGGFGGIGGFGGGMGGGPGGFGGMGGFGGMAAKGGFGGMGSGMGGFGGMGGGNAGFGGMGGGNAGFGGMGGQGGFGGKGY (SEQ ID NO:61).

[0197] In exemplary embodiments, the abductin or ALP comprises SEQ ID NO: 36, or a sequence having no more than one amino acid substitution compared to SEQ ID NO: 36. The abductin or ALP may comprise 1 to 10 sequences having SEQ ID NO: 36, or 1 to 10 sequences each independently having no more than one amino acid substitution compared to SEQ ID NO: 36. The 1 to 10 sequences may be linked, such as SEQ ID NO: 42, and separated by one or more amino acids.

[0198] In some embodiments, the fusion protein comprises a linker moiety. In some embodiments, the linker moiety is spacer-like. In some embodiments, the linker moiety is disposed between the keratin or keratin-associated peptide and the polypeptide moiety. In some embodiments, the linker moiety is disposed between each polypeptide moiety. In some embodiments, the linker or spacer moiety is (GQ) n wherein n is an integer from 1 to 20. In some embodiments, the linker or spacer comprises (AK) n wherein n is an integer from 1 to 20. In some embodiments, the linker or spacer is (LK) n and n is an integer from 1 to 20. In some embodiments, the linker or spacer is (GA) nwherein n is an integer between 1 and 20. In some embodiments, the linker or spacer-like comprises GQGQGQGQGQGQGQGQGQGQGQGQGQGQ (SEQ ID NO:97), AKAKAKAKAKAKAAKAKAKAKAK (SEQ ID NO:98), LKLKLKLKLKLKLKLKLKLKLK (SEQ ID NO:99), GAGAGAGAGAGAGAGAGAGA (SEQ ID NO:100), or a combination thereof. In some embodiments, the linker or spacer-like comprises a fragment of GQGQGQGQGQGQGQGQGQGQGQGQGQ (SEQ ID NO:97), AKAKAKAKAKAKAAKAKAKAK (SEQ ID NO:98), LKLKLKLKLKLKLKLKLKLKLKLK (SEQ ID NO:99), GAGAGAGAGAGAGAGAGAGA (SEQ ID NO:100), or a combination thereof. n wherein n is 1 to 20. In some embodiments, the linker or spacer comprises (AK) n wherein n is 1 to 20. In some embodiments, the linker or spacer is (LK) n and n is 1 to 20. In some embodiments, the linker or spacer is (GA) n where n is 1 to 20.

[0199] [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7]

[0200] Table 1 provides exemplary embodiments of elastin, elastin-like polypeptide (ELP), silk, silk-like polypeptide (SLP), collagen, collagen-like polypeptide (CLP), resilin, resilin-like polypeptide (RLP), abductin, and abductin polypeptide (ALP) peptides.

[0201] [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6] [Table 2-7] [Table 2-8] [Table 2-9] [Table 2-10] [Table 2-11] [Table 2-12] [Table 2-13]

[0202] Table 2 shows examples of keratins and keratin-related fusion proteins.

[0203] In some embodiments, the keratin or keratin-associated fusion protein / peptide comprises, or is selected from the list of sequences set forth in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7; SEQ ID NO:44; SEQ ID NO:45; SEQ ID NO:46; SEQ ID NO:47. In some embodiments, the keratin or keratin-associated fusion protein comprises, or is selected from any one of SEQ ID NOs:200-551.

[0204] In some embodiments, the keratin or keratin-related fusion protein / peptide is SEQ ID NO:1.

[0205] In some embodiments, the keratin or keratin-related fusion protein / peptide is SEQ ID NO:2.

[0206] In some embodiments, the keratin or keratin-associated fusion protein / peptide is SEQ ID NO:3.

[0207] In some embodiments, the keratin or keratin-related fusion protein / peptide is SEQ ID NO:4.

[0208] In some embodiments, the keratin or keratin-related fusion protein / peptide is SEQ ID NO:5.

[0209] In some embodiments, the keratin or keratin-related fusion protein / peptide is SEQ ID NO:6.

[0210] In some embodiments, the keratin or keratin-related fusion protein / peptide is SEQ ID NO:7.

[0211] In some embodiments, the keratin or keratin-related fusion protein / peptide is SEQ ID NO:44.

[0212] In some embodiments, the keratin or keratin-related fusion protein / peptide is SEQ ID NO:45.

[0213] In some embodiments, the keratin or keratin-related fusion protein / peptide is SEQ ID NO:46.

[0214] In some embodiments, the keratin or keratin-related fusion protein / peptide is SEQ ID NO:47.

[0215] In some embodiments, the keratin or keratin-related fusion protein / peptide comprises a protein having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or more sequence identity to any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, or SEQ ID NOs:200-551.

[0216] In some embodiments, the keratin or keratin-related fusion protein comprises a protein having at least 50% sequence identity to any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, or SEQ ID NOs:200-551.

[0217] In some embodiments, the keratin or keratin-related fusion protein comprises a protein having at least 55% sequence identity to any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, or SEQ ID NOs:200-551.

[0218] In some embodiments, the keratin or keratin-related fusion protein comprises a protein having at least 60% sequence identity to any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, or SEQ ID NOs:200-551.

[0219] In some embodiments, the keratin or keratin-related fusion protein comprises a protein having at least 65% sequence identity to any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, or SEQ ID NOs:200-551.

[0220] In some embodiments, the keratin or keratin-related fusion protein comprises a protein having at least 70% sequence identity to any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, or SEQ ID NOs:200-551.

[0221] In some embodiments, the keratin or keratin-related fusion protein comprises a protein having at least 75% sequence identity to any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, or SEQ ID NOs:200-551.

[0222] In some embodiments, the keratin or keratin-related fusion protein comprises a protein having at least 80% sequence identity to any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, or SEQ ID NOs:200-551.

[0223] In some embodiments, the keratin or keratin-related fusion protein comprises a protein having at least 85% sequence identity to any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, or SEQ ID NOs:200-551.

[0224] In some embodiments, the keratin or keratin-related fusion protein comprises a protein having at least 90% sequence identity to any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, or SEQ ID NOs:200-551. In some embodiments, the keratin or keratin-related fusion protein comprises a protein having at least 91% sequence identity to any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, or SEQ ID NOs:200-551. In some embodiments, the keratin or keratin-related fusion protein comprises a protein having at least 92% sequence identity to any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, or SEQ ID NOs:200-551. In some embodiments, the keratin or keratin-related fusion protein comprises a protein having at least 93% sequence identity to any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, or SEQ ID NOs:200-551.In some embodiments, the keratin or keratin-related fusion protein comprises a protein having at least 94% sequence identity to any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, or SEQ ID NOs:200-551.

[0225] In some embodiments, the keratin or keratin-related fusion protein comprises a protein having at least 95% sequence identity to any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, or SEQ ID NOs:200-551. In some embodiments, the keratin or keratin-related fusion protein comprises a protein having at least 96% sequence identity to any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, or SEQ ID NOs:200-551. In some embodiments, the keratin or keratin-related fusion protein comprises a protein having at least 97% sequence identity to any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, or SEQ ID NOs:200-551. In some embodiments, the keratin or keratin-related fusion protein comprises a protein having at least 98% sequence identity to any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, or SEQ ID NOs:200-551.

[0226] In some embodiments, the keratin or keratin-related fusion protein comprises a protein having at least 99% sequence identity to any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, or SEQ ID NOs:200-551.

[0227] In some embodiments, the keratin or keratin-associated fusion protein comprises a keratin or keratin-associated peptide linked to elastin or ELP. The linkage can be a covalent bond. The linkage can be via a linker. The linker can be a peptide linker. The peptide linker can comprise from about 1 to about 10 amino acids. The fusion protein can comprise GGVCGPSPPCITTVPGVG (SEQ ID NO: 62). The fusion protein can comprise any one of SEQ ID NOs: 1, 2, 7, or 200-271.

[0228] In some embodiments, the keratin or keratin-related fusion protein comprises a keratin or keratin-related peptide linked to a silk or SLP. The linkage can be a covalent bond. The linkage can be via a linker. The linker can be a peptide linker. The peptide linker can comprise from about 1 to about 10 amino acids. The fusion protein can comprise GGVCGPSPPCITTGAGAGS (SEQ ID NO: 63). The fusion protein can comprise any one of SEQ ID NOs: 3 or 272-315.

[0229] In some embodiments, the keratin or keratin-associated fusion protein comprises a keratin or keratin-associated peptide linked to resilin or RLP. The linkage can be a covalent bond. The linkage can be via a linker. The linker can be a peptide linker. The peptide linker can comprise from about 1 to about 10 amino acids. The fusion protein can comprise GGVCGPSPPCITTGGRPSDSYGAPGGGN (SEQ ID NO:4). The fusion protein can comprise any one of SEQ ID NOs: 4 or 375-418.

[0230] In some embodiments, the keratin or keratin-associated fusion protein comprises a keratin or keratin-associated peptide linked to collagen or CLP. The linkage can be a covalent bond. The linkage can be via a linker. The linker can be a peptide linker. The peptide linker can comprise from about 1 to about 10 amino acids. The fusion protein can comprise GGVCGPSPPCITTGPT (SEQ ID NO: 64). The fusion protein can comprise any one of SEQ ID NOs: 5 or 316-374.

[0231] In some embodiments, the keratin or keratin-associated fusion protein comprises a keratin or keratin-associated peptide linked to an abductin or ALP. The linkage can be a covalent bond. The linkage can be via a linker. The linker can be a peptide linker. The peptide linker can comprise from about 1 to about 10 amino acids. The fusion protein can comprise GGVCGPSPPCITTGGFGGMGGGS (SEQ ID NO: 65). The fusion protein can comprise any one of SEQ ID NOs: 6 or 419-477.

[0232] (Keratin and keratin-related fusion proteins / peptides) In other or additional embodiments, the present disclosure includes keratin and keratin-associated fusion proteins / peptides. The keratin and keratin-associated fusion proteins / peptides of the present disclosure comprise copolymers comprising various blocks. Each block of the copolymer comprises monomers of amino acid residues, such as alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, and two or more of these amino acids.

[0233] In some embodiments, the keratin and keratin-associated fusion proteins / peptides disclosed herein comprise amino acid sequences (the term "peptide" can be used interchangeably to refer to amino acid sequences), which can be joined by covalent bonds or any other type of bond that results in a structure that maintains the properties of the fusion proteins disclosed herein. The bond can be selected from, but is not limited to, hydrogen bonds, ion pairing, hydrophobic associations, or inclusion complex formation.

[0234] In some embodiments, keratin and keratin-associated fusion proteins are represented by the formula (I) [B n1 -A m -B n -C o -B n2 ] p It has.

[0235] In some embodiments, keratin and keratin-associated fusion proteins are represented by the formula (II) [B n1 -C o -B n -A m -B n2 ] p It has.

[0236] In some embodiments, keratin and keratin-associated fusion proteins are represented by the formula (III) [B n1 -B n -C o -Am -B n2 ] p It has.

[0237] In some embodiments, keratin and keratin-associated fusion proteins are represented by the formula (IV) [B n1 -C o -A m -B n -B n2 ] p It has.

[0238] In some embodiments, "A" comprises a peptide sequence comprising a keratin or keratin-associated peptide block. In exemplary embodiments, the keratin or keratin-associated peptide block is a keratin or keratin-associated peptide component described elsewhere herein. In some embodiments, the keratin, keratin peptide, or keratin-associated peptide comprises a wild-type keratin peptide, a functional fragment of a wild-type keratin peptide, or a peptide having at least 90% identity to a wild-type keratin peptide or a functional fragment thereof. The wild-type keratin peptide may be keratin 85 having SEQ ID NO:48. The wild-type keratin peptide may comprise SEQ ID NO:8, 9, 10, 66, or 67, or a combination of two or more thereof. In some embodiments, the keratin, keratin peptide, or keratin-associated peptide forms one or more disulfide bonds with hair, skin, or nails, or two or more thereof.

[0239] In non-limiting exemplary embodiments, a functional fragment of a wild-type keratin peptide comprises at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 consecutive amino acids of the wild-type keratin peptide. In non-limiting exemplary embodiments, a functional fragment of a wild-type keratin peptide comprises at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 amino acids that are at least 90% identical to the wild-type keratin peptide. In non-limiting exemplary embodiments, a functional fragment of a wild-type keratin peptide comprises at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 amino acids, wherein the at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 amino acids comprise one or more cysteine ​​amino acids. For example, the one or more cysteine ​​amino acids are 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or 27 cysteine ​​amino acids. In non-limiting exemplary embodiments, a functional fragment of a wild-type keratin peptide comprises at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 amino acids, wherein at least 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 30%, 40%, or 50% of the at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 amino acids are cysteine ​​amino acids. A non-limiting exemplary embodiment of a keratin or keratin peptide is shown in SEQ ID NO:48.

[0240] In some embodiments, keratin-associated peptides (sometimes referred to as keratin-associated proteins) include peptides that form disulfide bonds with wild-type keratin. In some embodiments, the keratin-associated peptides comprise at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 amino acids, and at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 amino acids are at least 90% identical to the wild-type keratin peptide sequence. The wild-type keratin peptide can be keratin 85, having SEQ ID NO: 48. The wild-type keratin peptide can comprise SEQ ID NO: 8, 9, 10, 66, or 67, or a combination of two or more thereof.

[0241] In some embodiments, the number of amino acids in the keratin or keratin-associated peptide block of the fusion protein / peptide is from about 6 to about 16 residues.

[0242] In some embodiments, the number of amino acids in the keratin or keratin-associated peptide block of the fusion protein / peptide is about 6 residues.

[0243] In some embodiments, the number of amino acids in the keratin or keratin-associated peptide block of the fusion protein / peptide is about 7 residues.

[0244] In some embodiments, the number of amino acids in the keratin or keratin-associated peptide block of the fusion protein / peptide is about 8 residues.

[0245] In some embodiments, the number of amino acids in the keratin or keratin-associated peptide block of the fusion protein / peptide is about 9 residues.

[0246] In some embodiments, the number of amino acids in the keratin or keratin-associated peptide block of the fusion protein / peptide is about 10 residues.

[0247] In some embodiments, the number of amino acids in the keratin or keratin-associated peptide block of the fusion protein / peptide is about 11 residues.

[0248] In some embodiments, the number of amino acids in the keratin or keratin-associated peptide block of the fusion protein / peptide is about 12 residues.

[0249] In some embodiments, the number of amino acids in the keratin or keratin-associated peptide block of the fusion protein / peptide is about 13 residues.

[0250] In some embodiments, the number of amino acids in the keratin or keratin-associated peptide block of the fusion protein / peptide is about 14 residues.

[0251] In some embodiments, the number of amino acids in the keratin or keratin-associated peptide block of the fusion protein / peptide is about 15 residues.

[0252] In some embodiments, the number of amino acids in the keratin or keratin-associated peptide block of the fusion protein / peptide is about 16 residues.

[0253] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein / peptide has a relatively high cysteine ​​content to enhance certain properties of the keratin or keratin-associated peptide block, such as intermolecular cross-linking. The total cysteine ​​content ranges from 10% to 50% of the total number of amino acids in the keratin or keratin-associated peptide block. In some embodiments, the total cysteine ​​content is at least 15% of the total number of amino acids in the keratin or keratin-associated peptide block.

[0254] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein comprises two cysteine ​​residues.

[0255] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein comprises three cysteine ​​residues.

[0256] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein comprises four cysteine ​​residues.

[0257] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein comprises five cysteine ​​residues.

[0258] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein comprises the peptide sequence (X 3 ) k C(X 3 )5C(X 3 ) k It has.

[0259] In some embodiments, each "X 3 " are independently glycine, alanine, valine, isoleucine, leucine, phenylalanine, proline, serine, threonine, asparagine, glutamine, cysteine, methionine, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, histidine, lysine, or arginine.

[0260] In some embodiments, each "k" is independently an integer from 0 to 3. When k is 2, (X 3 ) k has a first X3 and a second X3, and each X3 is independently glycine, alanine, valine, isoleucine, leucine, phenylalanine, proline, serine, threonine, asparagine, glutamine, cysteine, methionine, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, histidine, lysine, or arginine. When k is 3, (X 3 ) k is the first X 3 , the second X 3 and a third X 3 and each X3 are independently glycine, alanine, valine, isoleucine, leucine, phenylalanine, proline, serine, threonine, asparagine, glutamine, cysteine, methionine, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, histidine, lysine, or arginine. (X 3 )5 is the first X 3 , the second X 3 , the third X 3 , 4th X 3 and the fifth X 3 and each X 3 are independently glycine, alanine, valine, isoleucine, leucine, phenylalanine, proline, serine, threonine, asparagine, glutamine, cysteine, methionine, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, histidine, lysine, or arginine.

[0261] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein / peptide is selected from the group consisting of GGVCGPSPPCITT (SEQ ID NO:8), GGVCGPSPPC (SEQ ID NO:9), CGPSPPCITT (SEQ ID NO:10), CGPSPPC (SEQ ID NO:66), GVCGPSPPC (SEQ ID NO:67), CLPCLPAASC (SEQ ID NO:68), CLPAASC (SEQ ID NO:69), YGGSSGGG (SEQ ID NO:70), FGGGS (SEQ ID NO:71), CCQSSCCKPSC (SEQ ID NO:72), CVSSCCKPQCC (SEQ ID NO:73), PITCRRTCYH (SEQ ID NO:74), DCKLPCNPCA (SEQ ID NO:75), CLPCLPAASC (SEQ ID NO:76), CEPAICEPSC (SEQ ID NO:77), NO:77), CQCSCCKPYCS (SEQ ID NO:78), FCGFPSCSTSC (SEQ ID NO:79), CTPPSCCQLHHA (SEQ ID NO:80), SCCAPVYCCK (SEQ ID NO:81), or a combination of two or more thereof.

[0262] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein / peptide comprises GGVCGPSPPCITT (SEQ ID NO:8), GGVCGPSPPC (SEQ ID NO:9), CGPSPPCITT (SEQ ID NO:10), CGPSPPC (SEQ ID NO:66), or GVCGPSPPC (SEQ ID NO:67), or a combination of two or more thereof.

[0263] In some embodiments, the keratin or keratin-associated peptide block is GGVCGPSPPCITT (SEQ ID NO:8) or CGPSPPCITT (SEQ ID NO:10).

[0264] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein / peptide comprises YGGSSGGG (SEQ ID NO:70), FGGGS (SEQ ID NO:71), or a combination of two or more thereof.

[0265] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein / peptide comprises CCQSSCCKPSC (SEQ ID NO:72), CVSSCCKPQCC (SEQ ID NO:73), or a combination of two or more thereof.

[0266] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein / peptide comprises PITCRRTCYH (SEQ ID NO:74).

[0267] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein comprises DCKLPCNPCA (SEQ ID NO:75).

[0268] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein comprises CLPCLPAASC (SEQ ID NO:76).

[0269] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein comprises CEPAICEPSC (SEQ ID NO:77).

[0270] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein comprises CQCSCCKPYCS (SEQ ID NO:78).

[0271] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein comprises FCGFPSCSTSC (SEQ ID NO:79).

[0272] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein comprises CTPPSCCQLHHA (SEQ ID NO:80).

[0273] In some embodiments, the keratin or keratin-associated peptide block of the fusion protein comprises SCCAPVYCCK (SEQ ID NO:81).

[0274] In some embodiments, "m" is an integer from 1 to 10.

[0275] In some embodiments, "m" is about 1.

[0276] In some embodiments, "m" is about 2.

[0277] In some embodiments, "m" is about 3.

[0278] In some embodiments, "m" is about 4.

[0279] In some embodiments, "m" is about 5.

[0280] In some embodiments, "m" is about 6.

[0281] In some embodiments, "m" is about 7.

[0282] In some embodiments, "m" is about 8.

[0283] In some embodiments, "m" is about 9.

[0284] In some embodiments, "m" is about 10.

[0285] In some embodiments, "B" comprises a linker or terminus. Linkers aid in fusion protein design by providing appropriate spacing between domains and support correct protein folding when N- or C-terminal interactions are important for folding. Generally, linkers allow important interdomain interactions, enhance stability, flexibility, and reduce steric hindrance, making them suitable for use in protein / peptide design even when the N- and C-termini may be fused. The three main types of linkers are flexible, rigid, and in vivo cleavable. Terminal (N- or C-terminal) peptides aid in the conjugation process, stability during synthesis, and correct folding.

[0286] In some embodiments, "B" comprises a peptide sequence comprising or having 1 to 3 amino acids. In some embodiments, "B" comprises a peptide sequence comprising or having 1 to 3 amino acids repeated 0 to about 25 times. In some embodiments, "B" comprises a peptide sequence comprising 1 or 2 amino acids repeated about 10 to about 25 times. In some embodiments, each amino acid in the peptide sequence is independently arginine, histidine, lysine, aspartic acid, glutamic acid, serine, threonine, asparagine, glutamine, cysteine, selenocysteine, glycine, proline, alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, or tryptophan. In some embodiments, B has 1 to 40 amino acids.

[0287] In some embodiments, each "n," "n1," and "n2" is independently an integer from 0 to 25.

[0288] In some embodiments, "n, n1 and n2" are 0.

[0289] In some embodiments, "n, n1 and n2" are about 1.

[0290] In some embodiments, "n, n1 and n2" are about 2.

[0291] In some embodiments, "n, n1 and n2" are about 3.

[0292] In some embodiments, "n, n1 and n2" are about 4.

[0293] In some embodiments, "n, n1 and n2" are about 5.

[0294] In some embodiments, "n, n1 and n2" are about 6.

[0295] In some embodiments, "n, n1 and n2" are about 7.

[0296] In some embodiments, "n, n1 and n2" are about 8.

[0297] In some embodiments, "n, n1 and n2" are about 9.

[0298] In some embodiments, "n, n1 and n2" are about 10.

[0299] In some embodiments, "n, n1 and n2" are about 11.

[0300] In some embodiments, "n, n1 and n2" are about 12.

[0301] In some embodiments, "n, n1 and n2" are about 13.

[0302] In some embodiments, "n, n1 and n2" are about 14.

[0303] In some embodiments, "n, n1 and n2" are about 15.

[0304] In some embodiments, "n, n1 and n2" are about 16.

[0305] In some embodiments, "n, n1 and n2" are about 17.

[0306] In some embodiments, "n, n1 and n2" are about 18.

[0307] In some embodiments, "n, n1 and n2" are about 19.

[0308] In some embodiments, "n, n1 and n2" are about 20.

[0309] In some embodiments, "n, n1 and n2" are about 21.

[0310] In some embodiments, "n, n1 and n2" are about 22.

[0311] In some embodiments, "n, n1 and n2" are about 23.

[0312] In some embodiments, "n, n1 and n2" are about 24.

[0313] In some embodiments, "n, n1 and n2" are about 25.

[0314] In some embodiments, "C" comprises a peptide sequence having the structure Dq-Er-Fs-Gt-Hu.

[0315] In some embodiments, blocks "D", "E", "F", "G" and "H" may be provided in any order.

[0316] In some embodiments, "D" comprises a silk or silk-like peptide (SLP) block.

[0317] In some embodiments, "E" comprises a collagen or collagen-like peptide (CLP) block.

[0318] In some embodiments, "F" comprises an elastin or elastin-like peptide (ELP) block.

[0319] In some embodiments, "G" comprises a resilin or resilin-like peptide (RLP) block.

[0320] In some embodiments, "H" comprises an abductin or abductin-like peptide (ALP) block.

[0321] In some embodiments, the keratin or keratin-related fusion proteins disclosed herein comprise an elastin or elastin-like polypeptide (ELP) block ("F"). In some embodiments, elastin-like polypeptide (ELP) refers to a peptide (including a synthetic peptide) comprising an amino acid sequence having a similar secondary structure, tertiary structure, or amino acid sequence, or a combination of two or more thereof, to a wild-type elastin protein (e.g., SEQ ID NO: 55) or a fragment of a wild-type elastin protein. In some embodiments, the fragment comprises a sequence having 4, 5, 6, 7, 8, 9, or 10 amino acids that are 100% identical to the sequence of the wild-type elastin protein. Non-limiting examples of elastins and ELPs include SEQ ID NOs: 11-23, 49-55, 82-91, and 104.

[0322] The ELP may comprise a polymeric or oligomeric repeating peptide sequence of elastin protein. In exemplary embodiments, the elastin or ELP comprises SEQ ID NO:49 ((VPGXG)n, where n is 1-10 and each X is independently any amino acid), or a sequence having no more than one amino acid substitution compared to SEQ ID NO:49. In some embodiments, each X is independently valine or leucine. In some embodiments, n is 5. In some embodiments, n is 6.

[0323] In exemplary embodiments, the elastin or ELP comprises SEQ ID NO:50 (VPGVG), or a sequence having no more than one amino acid substitution compared to SEQ ID NO:50. The elastin or ELP may comprise 1 to 10 sequences having SEQ ID NO:50, or 1 to 10 sequences each independently having no more than one amino acid substitution compared to SEQ ID NO:50. The 1 to 10 sequences may be linked, e.g., VPGVGVPGVGVPGVGVPGVGVPGVG (SEQ ID NO:23), or separated by one or more amino acids by another ELP, e.g., VPGVGVPGVGVPGLGVPGVGVPGVG (SEQ ID NO:54).

[0324] In exemplary embodiments, the elastin or ELP comprises SEQ ID NO:51 (VPGLG), or a sequence having no more than one amino acid substitution compared to SEQ ID NO: 51. The elastin or ELP may comprise 1 to 10 sequences having SEQ ID NO:51, or 1 to 10 sequences each independently having no more than one amino acid substitution compared to SEQ ID NO:51. The 1 to 10 sequences may be linked or separated by one or more amino acids.

[0325] In exemplary embodiments, the elastin or ELP comprises SEQ ID NO:52 (VPGVGVPGL), or a sequence having no more than one amino acid substitution compared to SEQ ID NO:52. The elastin or ELP may comprise 1 to 10 sequences having SEQ ID NO:52, or 1 to 10 sequences each independently having no more than one amino acid substitution compared to SEQ ID NO:52. The 1 to 10 sequences may be linked or separated by one or more amino acids.

[0326] In exemplary embodiments, the elastin or ELP comprises SEQ ID NO: 53 (VPGVGVPGVGVPGLGVPGVGVPGVGR), or a sequence having no more than one amino acid substitution compared to SEQ ID NO: 53. The elastin or ELP may comprise 1 to 10 sequences each independently having SEQ ID NO: 53, or a sequence having no more than one amino acid substitution compared to SEQ ID NO: 53. The 1 to 10 sequences may be linked or separated by one or more amino acids.

[0327] In exemplary embodiments, the elastin or ELP comprises SEQ ID NO:23 (VPGVGVPGVGVPGVGVPGVGVPGVG), or a sequence having no more than one amino acid substitution compared to SEQ ID NO: 23. The elastin or ELP may comprise 1 to 10 sequences each independently having SEQ ID NO:23, or a sequence having no more than one amino acid substitution compared to SEQ ID NO:23. The 1 to 10 sequences may be linked or separated by one or more amino acids.

[0328] In exemplary embodiments, the elastin or ELP comprises SEQ ID NO:54 (VPGVGVPGVGVPGLGVPGVGVPGVG), or a sequence having no more than one amino acid substitution compared to SEQ ID NO:54. The elastin or ELP may comprise 1 to 10 sequences each independently having SEQ ID NO:54, or a sequence having no more than one amino acid substitution compared to SEQ ID NO:54. The 1 to 10 sequences may be linked or separated by one or more amino acids.

[0329] In exemplary embodiments, the elastin or ELP comprises SEQ ID NO:22 (GVGVPGVGVPGLGVPGVGVPGVG), or a sequence having no more than one amino acid substitution compared to SEQ ID NO: 22. The elastin or ELP may comprise 1 to 10 sequences each independently having SEQ ID NO:22, or a sequence having no more than one amino acid substitution compared to SEQ ID NO:22. The 1 to 10 sequences may be linked or separated by one or more amino acids.

[0330] In exemplary embodiments, the elastin or ELP comprises SEQ ID NO:82 (IPGLG), or a sequence having no more than one amino acid substitution compared to SEQ ID NO:82. The elastin or ELP may comprise 1 to 10 sequences having SEQ ID NO:82, or 1 to 10 sequences each independently having no more than one amino acid substitution compared to SEQ ID NO:82. The 1 to 10 sequences may be concatenated or separated by one or more amino acids. In some embodiments, the elastin or ELP comprises SEQ ID NO:85 (IPGVGIPGLG), SEQ ID NO:89 (VPGVGIPGLG), or a combination thereof.

[0331] In some embodiments, the elastin or ELP comprises polymeric or oligomeric repeating peptide sequences of elastin peptides, hi some embodiments, each polymeric or oligomeric repeating peptide sequence comprises or has 4 amino acid residues.

[0332] In some embodiments, each polymeric or oligomeric repeating peptide sequence comprises or has 5 amino acid residues. Non-limiting examples include SEQ ID NOs: 49, 50, or 51.

[0333] In some embodiments, each polymeric or oligomeric repeating peptide sequence comprises or has 6 amino acid residues.

[0334] In some embodiments, each polymeric or oligomeric repeat peptide sequence comprises or has 7 amino acid residues.

[0335] In some embodiments, each polymeric or oligomeric repeat peptide sequence comprises or has 8 amino acid residues.

[0336] Non-limiting examples of repeating peptide sequences include polytetrapeptides, polypentapeptides, polyhexapeptides, polyheptapeptides, polyoctapeptides, and polynonapeptides, including VPGG (SEQ ID NO:11), IPGG (SEQ ID NO:12), VPAVG (SEQ ID NO:13), AVGVP (SEQ ID NO:14), IPGVG (SEQ ID NO:15), LPGVG (SEQ ID NO:16), VAPGVG (SEQ ID NO:17), GVGVPGVG (SEQ ID NO:18), VPGFGVGAG (SEQ ID NO:19), VPGVGVPGG (SEQ ID NO:20), VPGX2G (SEQ ID NO:21), SEQ ID NO:49 ((VPGXG)n, where n is 1-10 and each X is independently any amino acid), SEQ ID NO:50 (VPGVG), SEQ ID NO:51 (VPGVG), SEQ ID NO:52 (VPGVG), SEQ ID NO:53 (VPGVG), SEQ ID NO:54 (VPGVG), SEQ ID NO:55 (VPGVG), SEQ ID NO:56 (VPGVG), SEQ ID NO:57 (VPGVG), SEQ ID NO:58 (VPGVG), SEQ ID NO:59 (VPGVG), SEQ ID NO:60 (VPGVG), SEQ ID NO:61 (VPGVG), SEQ ID NO:62 (VPGVG), SEQ ID NO:63 (VPGVG), SEQ ID NO:64 (VPGVG), SEQ ID NO:65 (VPGVG), SEQ ID NO:66 (VPGVG), SEQ ID NO:67 (VPGVG), SEQ ID NO:68 (VPGVG), SEQ ID NO:69 (VPGVG), SEQ ID NO:70 (VPGVG), SEQ ID NO:71 (VPGVG), SEQ ID NO:72 (VPGVG), SEQ ID NO:73 (VPG SEQ ID NO:51 (VPGLG), and SEQ ID NO:52 (VPGVGVPGL), and combinations of two or more thereof.

[0337] In some embodiments, "X 2 " comprises naturally occurring or non-naturally occurring amino acids.

[0338] In some embodiments, the elastin or ELP is a polymeric or oligomeric repeat of the pentapeptide VPAVG.

[0339] In some embodiments, elastin or ELP is the pentapeptide VPGX 2 G is a polymer or oligomer of repeating "X 2" is an amino acid selected from arginine, histidine, lysine, aspartic acid, glutamic acid, serine, threonine, asparagine, glutamine, cysteine, glycine, alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, or tryptophan. In some embodiments, "X 2 " is leucine. In some embodiments, "X 2 " is asparagine.

[0340] In some embodiments, "X 2 " is Balin.

[0341] In some embodiments, elastin or ELP is defined by the peptide sequence GVGVPGVGVPGLGVPGVGVPGVG (SEQ ID NO:22).

[0342] In some embodiments, elastin or ELP is defined by the sequence VPGVGVPGVGVPGVGVPGVGVPGVG (SEQ ID NO:23).

[0343] In some embodiments, the elastin or elastin-like polypeptide (ELP) block forms a beta-sheet rich domain.

[0344] In some embodiments, the elastin or ELP block comprises an amorphous domain. In some embodiments, the elastin or ELP block comprises a crystalline domain. In some embodiments, the elastin or ELP block comprises both crystalline and amorphous domains.

[0345] In some embodiments, the elastin or ELP block is stimulus-responsive. In some embodiments, the ELP block is temperature-responsive. In some embodiments, the ELP component may respond to pH changes. In some embodiments, the ELP component may respond over a range of temperatures. In some embodiments, the ELP component may respond to salt.

[0346] In some embodiments, "s" is an integer from 0 to about 220.

[0347] In some embodiments, "s" is 0.

[0348] In some embodiments, "s" is about 1.

[0349] In some embodiments, "s" is about 2.

[0350] In some embodiments, "s" is about 3.

[0351] In some embodiments, "s" is about 4.

[0352] In some embodiments, "s" is about 5.

[0353] In some embodiments, "s" is about 6.

[0354] In some embodiments, "s" is about 7.

[0355] In some embodiments, "s" is about 8.

[0356] In some embodiments, "s" is about 9.

[0357] In some embodiments, "s" is about 10.

[0358] In some embodiments, "s" is about 20.

[0359] In some embodiments, "s" is about 30.

[0360] In some embodiments, "s" is about 40.

[0361] In some embodiments, "s" is about 50.

[0362] In some embodiments, "s" is about 100.

[0363] In some embodiments, the keratin or keratin-related fusion proteins / peptides disclosed herein comprise a silk or silk-like polypeptide (SLP) block ("D"). In some embodiments, a silk-like polypeptide (SLP) refers to a peptide, including a synthetic peptide having a similar secondary structure, tertiary structure, or amino acid sequence to a wild-type silk protein (e.g., SEQ ID NO: 56) or a fragment of a wild-type silk protein, or a combination of two or more thereof. In some embodiments, the fragment comprises a sequence having at least 4, 5, 6, 7, 8, 9, or 10 amino acids that are 100% identical to the sequence of the wild-type silk protein. The wild-type silk protein can be fibroin. Non-limiting examples of silks and SLPs include SEQ ID NOs: 24-32, 92, and 56.

[0364] The SLP may comprise a polymeric or oligomeric repeating peptide sequence of a silk protein. In one exemplary embodiment, the silk or SLP comprises SEQ ID NO:24 GAGAGS, or a sequence having up to one amino acid substitution compared to SEQ ID NO:24. In one exemplary embodiment, the silk or SLP comprises SEQ ID NO:24, or a sequence having up to one amino acid substitution compared to SEQ ID NO:24. The silk or SLP may comprise 1 to 10 sequences having SEQ ID NO:24, or 1 to 10 sequences each independently having up to one amino acid substitution compared to SEQ ID NO:24. The 1 to 10 sequences may be linked or separated by one or more amino acids.

[0365] In some embodiments, the silk or SLP comprises polymeric or oligomeric repeating peptide sequences of silk peptides, each polymeric or oligomeric repeating peptide sequence comprising 6 amino acid residues.

[0366] In some embodiments, the silk or SLP comprises polymeric or oligomeric repeating peptide sequences of silk peptides, each polymeric or oligomeric repeating peptide sequence comprising 7 amino acid residues.

[0367] In some embodiments, the silk or SLP comprises polymeric or oligomeric repeating peptide sequences of silk peptides, each polymeric or oligomeric repeating peptide sequence comprising 8 amino acid residues.

[0368] Non-limiting examples of repeat peptide sequences include polytetrapeptides, polypentapeptides, polyhexapeptides, polyheptapeptides, polyoctapeptides, and polynonapeptides, including, but not limited to, GAGAGS (SEQ ID NO:24), GAGSGA (SEQ ID NO:25), GAGAGY (SEQ ID NO:26), GAGYGA (SEQ ID NO:27), GAGAGA (SEQ ID NO:28), GAGAGV (SEQ ID NO:29), GAGVGA (SEQ ID NO:30), GAGAGVGY (SEQ ID NO:31), and combinations of two or more thereof.

[0369] In some embodiments, silks or SLPs are polymeric or oligomeric repeats of the pentapeptide GAGAGS (SEQ ID NO:24).

[0370] In some embodiments, the silk or SLP is defined by the peptide sequence GAGAGSGAGAGSGAGAGSGAGAGSGAGAGS (SEQ ID NO:32).

[0371] In some embodiments, the silk or silk-like (SLP) blocks form a beta-sheet rich domain.

[0372] In some embodiments, the silk or SLP blocks comprise crystalline domains. In some embodiments, the silk or SLP blocks comprise amorphous domains. In some embodiments, the silk or SLP blocks comprise both amorphous and crystalline domains.

[0373] In some embodiments, "q" is an integer between 0 and 100.

[0374] In some embodiments, "q" is 0.

[0375] In some embodiments, "q" is about 1.

[0376] In some embodiments, "q" is about 2.

[0377] In some embodiments, "q" is about 3.

[0378] In some embodiments, "q" is about 4.

[0379] In some embodiments, "q" is about 5.

[0380] In some embodiments, "q" is about 6.

[0381] In some embodiments, "q" is about 7.

[0382] In some embodiments, "q" is about 8.

[0383] In some embodiments, "q" is about 9.

[0384] In some embodiments, "q" is about 10.

[0385] In some embodiments, "q" is about 20.

[0386] In some embodiments, "q" is about 30.

[0387] In some embodiments, "q" is about 40.

[0388] In some embodiments, "q" is about 50.

[0389] In some embodiments, "q" is about 100.

[0390] In some embodiments, the keratin or keratin-related fusion proteins / peptides disclosed herein comprise a collagen or collagen-like polypeptide (CLP) block ("E"). In some embodiments, collagen-like polypeptide (CLP) refers to a peptide, including a synthetic peptide having a similar secondary structure, tertiary structure, or amino acid sequence, or a combination of two or more thereof, to a wild-type collagen protein (e.g., SEQ ID NO: 59) or a fragment of a wild-type collagen protein. In some embodiments, the fragment comprises a sequence having at least 3, 4, 5, 6, 7, 8, 9, or 10 amino acids that are 100% identical to the sequence of the wild-type collagen protein. Non-limiting examples of collagens and CLPs include SEQ ID NOs: 33, 58, 59, 93, 94, 95, 57, and fragments having the amino acids "GTP" and / or "GLQ".

[0391] The CLP may comprise a polymeric or oligomeric repeating peptide sequence of collagen peptide. In one exemplary embodiment, the collagen or CLP comprises the sequence GTP. The collagen or CLP may comprise 1 to 10 sequences comprising the sequence GTP. The 1 to 10 sequences may be linked or separated by one or more amino acids. The 1 to 10 sequences may be, for example, two or three. For example, the collagen or CLP may comprise GPTGPT (SEQ ID NO: 57) or GPTGPTGPAGPRGLQGLQGLQGERGEQGPT (SEQ ID NO: 33). In one exemplary embodiment, the collagen or CLP may comprise the sequence GLQ. The collagen or CLP may comprise 1 to 10 sequences comprising the sequence GLQ. The 1 to 10 sequences may be, for example, two or three. The 1 to 10 sequences may be linked or separated by one or more amino acids. For example, the collagen or CLP may comprise GLQGLQ (SEQ ID NO:58) or GPTGPTGPAGPRGLQGLQGLQGERGEQGPT (SEQ ID NO:33).

[0392] In some embodiments, each polymeric or oligomeric repeating peptide sequence of the collagen or collagen-like polypeptide (CLP) block comprises three amino acid residues. As a non-limiting example, the three amino acid residues are GTP, GLQ, GX 1 It may contain H.

[0393] In some embodiments, the repeating peptide sequence comprises a polytetrapeptide, a polypentapeptide, a polyhexapeptide, a polyheptapeptide, a polyoctapeptide, a polynonapeptide, including GX 1 These include, but are not limited to, H.

[0394] In some embodiments, "X 1" comprises proline, leucine, isoleucine, glutamic acid, aspartic acid, glutamine, lysine, alanine, arginine and threonine, and "H" comprises an amino acid selected from glycine, alanine, lysine, proline, methionine, glutamine, arginine, threonine, valine, glutamic acid, aspartic acid or asparagine.

[0395] In some embodiments, the collagen or CLP is defined by the peptide sequence GPTGPTGPAGPRGLQGLQGLQGERGEQGPT (SEQ ID NO:33).

[0396] In some embodiments, "r" is an integer from 0 to about 200.

[0397] In some embodiments, "r" is 0.

[0398] In some embodiments, "r" is about 1.

[0399] In some embodiments, "r" is about 2.

[0400] In some embodiments, "r" is about 3.

[0401] In some embodiments, "r" is about 4.

[0402] In some embodiments, "r" is about 4.

[0403] In some embodiments, "r" is about 5.

[0404] In some embodiments, "r" is about 6.

[0405] In some embodiments, "r" is about 7.

[0406] In some embodiments, "r" is about 8.

[0407] In some embodiments, "r" is about 9.

[0408] In some embodiments, "r" is about 10.

[0409] In some embodiments, "r" is about 20.

[0410] In some embodiments, "r" is about 30.

[0411] In some embodiments, "r" is about 40.

[0412] In some embodiments, "r" is about 50.

[0413] In some embodiments, "r" is about 100.

[0414] In some embodiments, the keratin or keratin-associated fusion proteins / peptides disclosed herein comprise a resilin or resilin-like polypeptide (RLP) block ("G"). In some embodiments, resilin-like polypeptide (RLP) refers to a peptide, including a synthetic peptide having a similar secondary structure, tertiary structure, or amino acid sequence, or a combination of two or more thereof, to a wild-type resilin protein (e.g., SEQ ID NO: 60) or a fragment of a wild-type resilin protein. In some embodiments, the fragment comprises a sequence having at least 4, 5, 6, 7, 8, 9, or 10 amino acids that are 100% identical to the sequence of the wild-type silk protein. Non-limiting examples of resilin and RLPs include SEQ ID NOs: 34-35, 96, and 60.

[0415] The RLP comprises a polymeric or oligomeric repeating peptide sequence of the resilin protein. In one exemplary embodiment, the polymeric or oligomeric repeating peptide sequence is a fragment of the wild-type resilin peptide having about 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids.

[0416] In some embodiments, each polymeric or oligomeric repeat peptide sequence of a resilin or resilin-like polypeptide (RLP) component comprises 11 amino acid residues.

[0417] In some embodiments, each polymeric or oligomeric repeat peptide sequence of a resilin or resilin-like polypeptide (RLP) component comprises 12 amino acid residues.

[0418] In some embodiments, each polymeric or oligomeric repeat peptide sequence of a resilin or resilin-like polypeptide (RLP) component comprises 13 amino acid residues.

[0419] In some embodiments, each polymeric or oligomeric repeat peptide sequence of a resilin or resilin-like polypeptide (RLP) component comprises 14 amino acid residues.

[0420] In some embodiments, each polymeric or oligomeric repeat peptide sequence of a resilin or resilin-like polypeptide (RLP) component comprises 15 amino acid residues.

[0421] In some embodiments, exemplary repeating peptide sequences include polytetrapeptides, polypentapeptides, polyhexapeptides, polyheptapeptides, polyoctapeptides, polynonapeptides, including, but not limited to, GGRPSDSYGAPGGGN (SEQ ID NO:34), GAPAQTPSSQY (SEQ ID NO:35), AQTPSSQYGAP (SEQ ID NO:96), and combinations thereof.

[0422] In some embodiments, resilin or RLP is defined by the peptide sequence GGRPSDSYGAPGGGN (SEQ ID NO:34).

[0423] In some embodiments, "t" is an integer from 0 to about 80.

[0424] In some embodiments, "t" is 0.

[0425] In some embodiments, "t" is about 1.

[0426] In some embodiments, "t" is about 2.

[0427] In some embodiments, "t" is about 3.

[0428] In some embodiments, "t" is about 4.

[0429] In some embodiments, "t" is about 5.

[0430] In some embodiments, "t" is about 6.

[0431] In some embodiments, "t" is about 7.

[0432] In some embodiments, "t" is about 8.

[0433] In some embodiments, "t" is about 9.

[0434] In some embodiments, "t" is about 10.

[0435] In some embodiments, "t" is about 20.

[0436] In some embodiments, "t" is about 30.

[0437] In some embodiments, "t" is about 40.

[0438] In some embodiments, "t" is about 50.

[0439] In some embodiments, "t" is about 80.

[0440] In some embodiments, "t" is about 100.

[0441] In some embodiments, the keratin or keratin-associated component disclosed herein comprises an abductin or abductin-like polypeptide (ALP) block ("H"). In some embodiments, an abductin-like polypeptide (ALP) refers to a synthetic peptide comprising an amino acid sequence having a similar secondary structure, tertiary structure, or amino acid sequence to a wild-type abductin protein (e.g., SEQ ID NO:61) or a fragment of the wild-type abductin protein, or a combination of two or more thereof. In some embodiments, the fragment comprises a sequence having at least 4, 5, 6, 7, 8, 9, or 10 amino acids that are 100% identical to the sequence of the wild-type abductin protein. Non-limiting examples of abductins and ALPs include SEQ ID NOs:36-42, 61, and 108. An ALP comprises a polymeric or oligomeric repeating peptide sequence of an abductin protein.

[0442] In some embodiments, each polymeric or oligomeric repeat peptide sequence comprises four amino acid residues of an abductin protein.

[0443] In some embodiments, each polymeric or oligomeric repeat peptide sequence comprises five amino acid residues of an abductin protein.

[0444] In some embodiments, each polymeric or oligomeric repeat peptide sequence comprises 6 amino acid residues of an abductin protein.

[0445] In some embodiments, each polymeric or oligomeric repeat peptide sequence comprises 7 amino acid residues of an abductin protein.

[0446] In some embodiments, each polymeric or oligomeric repeat peptide sequence comprises 8 amino acid residues of an abductin protein.

[0447] In some embodiments, each polymeric or oligomeric repeat peptide sequence comprises 9 amino acid residues of an abductin protein.

[0448] In some embodiments, each polymeric or oligomeric repeat peptide sequence comprises 10 amino acid residues of an abductin protein.

[0449] In some embodiments, the repeating peptide sequences include polytetrapeptides, polypentapeptides, polyhexapeptides, polyheptapeptides, polyoctapeptides, and polynonapeptides, and are selected from the group consisting of GGFGGMGGGS (SEQ ID NO:36), MGGG (SEQ ID NO:37), FGGMG (SEQ ID NO:38), FGGMGGG (SEQ ID NO:39), GGFGGMGGG (SEQ ID NO:40), FGGMGGGNAG (SEQ ID NO:41), MNAYICLAACLIAAVSAAGYGGGAGSMGGTGGMGGGGMNAGGFGGMGGMGGGKGGFGGIGGFGGGMGGGPGGFGGMGGFGGMAAKGGFGGMGSGMGGFGGMGGGNAGFGGMGGGNAGFGGMGGQGGFGGKGY (SEQ ID NO:42), NO: 61) YGAPGGGNGGRPSDSYGAPGGGNGGRPSDSYGAPGGGNGGRPSDSYGAPGGGNGGRPSDSYGAPGGGNGQGQGQGQGQGQGQGQGQGGVCGPSPPCITTGQGQGQGQGQGQGQGQVPAVG VPAVGVPAVGVPAVGVPAVGVPAVGVPAVGVPAVGVPAVGVPAVGVPAVGGQGQGQGQGQGQGQGGVCGPSPPCITTGQGQGQGQGQGQGQGQGGRPSDSYGAPGGGNGGRPSDSYGAPGGGNGGRPSDSYGAPGGGNGGRPSDSYGAPGGGNGGRPSDSYGAPGGGNGGRPSDSYGAPGGGNGGRPSDSYGAPGGGNGGRPSDSYGAPGGGNGGRPSDSYGAPGGGNGGRPSDSYGAPGGGNGGRPSDSYGAPGGGN (SEQ ID NO: 108), and combinations thereof.

[0450] In some embodiments, the abductin or ALP is defined by the peptide sequence GGFGGMGGGSGGFGGMGGGSGGFGGMGGGS (SEQ ID NO:42). In exemplary embodiments, the abductin or ALP comprises SEQ ID NO:36, or a sequence having no more than one amino acid substitution compared to SEQ ID NO:36. The abductin or ALP may comprise 1 to 10 sequences having SEQ ID NO:36, or 1 to 10 sequences each having no more than one amino acid substitution compared to SEQ ID NO:36. The 1 to 10 sequences may be linked, for example, as in SEQ ID NO:42, or separated by one or more amino acids.

[0451] In some embodiments, "u" is an integer from 0 to about 100.

[0452] In some embodiments, "u" is 0 to about 70.

[0453] In some embodiments, "u" is 0.

[0454] In some embodiments, "u" is about 1.

[0455] In some embodiments, "u" is about 2.

[0456] In some embodiments, "u" is about 3.

[0457] In some embodiments, "u" is about 4.

[0458] In some embodiments, "u" is about 5.

[0459] In some embodiments, "u" is about 6.

[0460] In some embodiments, "u" is about 7.

[0461] In some embodiments, "u" is about 8.

[0462] In some embodiments, "u" is about 9.

[0463] In some embodiments, "u" is about 10.

[0464] In some embodiments, "u" is about 20.

[0465] In some embodiments, "u" is about 30.

[0466] In some embodiments, "u" is about 40.

[0467] In some embodiments, "u" is about 50.

[0468] In some embodiments, "u" is about 70.

[0469] In some embodiments, "o" is an integer from 0-30.

[0470] In some embodiments, "o" is 0.

[0471] In some embodiments, "o" is about 1.

[0472] In some embodiments, "o" is about 2.

[0473] In some embodiments, "o" is about 3.

[0474] In some embodiments, "o" is about 4.

[0475] In some embodiments, "o" is about 5.

[0476] In some embodiments, "o" is about 6.

[0477] In some embodiments, "o" is about 7.

[0478] In some embodiments, "o" is about 8.

[0479] In some embodiments, "o" is about 9.

[0480] In some embodiments, "o" is about 10.

[0481] In some embodiments, "o" is about 15.

[0482] In some embodiments, "o" is about 20.

[0483] In some embodiments, "o" is about 25.

[0484] In some embodiments, "o" is about 30.

[0485] In some embodiments, "p" is an integer from 1 to 30.

[0486] In some embodiments, "p" is about 1.

[0487] In some embodiments, "p" is about 2.

[0488] In some embodiments, "p" is about 3.

[0489] In some embodiments, "p" is about 4.

[0490] In some embodiments, "p" is about 5.

[0491] In some embodiments, "p" is about 6.

[0492] In some embodiments, "p" is about 7.

[0493] In some embodiments, "p" is about 8.

[0494] In some embodiments, "p" is about 9.

[0495] In some embodiments, "p" is about 10.

[0496] In some embodiments, "p" is about 15.

[0497] In some embodiments, "p" is about 20.

[0498] In some embodiments, "p" is about 25.

[0499] In some embodiments, "p" is about 30.

[0500] In some embodiments, the keratin or keratin-associated fusion protein / peptide has an average molecular weight range of about 1 kDa to about 144 kDa.

[0501] In some embodiments, the keratin or keratin-associated fusion protein / peptide has an average molecular weight range of about 1 kDa to about 5 kDa.

[0502] In some embodiments, the keratin or keratin-associated fusion protein / peptide has an average molecular weight range of about 5 kDa to about 10 kDa.

[0503] In some embodiments, the keratin or keratin-associated fusion protein / peptide has an average molecular weight range of about 6 kDa to about 17 kDa.

[0504] In some embodiments, the keratin or keratin-associated fusion protein / peptide has an average molecular weight range of about 10 kDa to about 15 kDa.

[0505] In some embodiments, the keratin or keratin-associated fusion protein / peptide has an average molecular weight range of about 14 kDa to about 30 kDa.

[0506] In some embodiments, the keratin or keratin-associated fusion protein / peptide has an average molecular weight range of about 15 kDa to about 20 kDa.

[0507] In some embodiments, the keratin or keratin-associated fusion protein / peptide has an average molecular weight range of about 17 kDa to about 39 kDa.

[0508] In some embodiments, the keratin or keratin-associated fusion protein / peptide may have an average molecular weight range of about 20 kDa to about 25 kDa.

[0509] In some embodiments, the keratin or keratin-associated fusion protein / peptide has an average molecular weight range of about 25 kDa to about 30 kDa.

[0510] In some embodiments, the keratin or keratin-associated fusion protein / peptide has an average molecular weight range of about 30 kDa to about 35 kDa.

[0511] In some embodiments, the keratin or keratin-associated fusion protein / peptide has an average molecular weight range of about 35 kDa to about 40 kDa.

[0512] In some embodiments, the keratin or keratin-associated fusion protein / peptide has an average molecular weight range of about 39 kDa to about 54 kDa.

[0513] In some embodiments, the keratin or keratin-associated fusion protein / peptide has an average molecular weight range of about 39 kDa to about 80 kDa.

[0514] In some embodiments, the keratin or keratin-associated fusion protein / peptide has an average molecular weight range of about 40 kDa to about 45 kDa.

[0515] In some embodiments, the keratin or keratin-associated fusion protein / peptide has an average molecular weight range of about 45 kDa to about 50 kDa.

[0516] In some embodiments, the keratin or keratin-associated fusion protein / peptide has an average molecular weight range of about 50 kDa to about 55 kDa.

[0517] In some embodiments, the keratin or keratin-associated fusion protein / peptide has an average molecular weight range of about 55 kDa to about 60 kDa.

[0518] In some embodiments, the keratin or keratin-associated fusion protein / peptide has an average molecular weight range of about 60 kDa to about 100 kDa.

[0519] In some embodiments, the keratin or keratin-associated fusion protein / peptide has an average molecular weight range of about 80 kDa to about 144 kDa.

[0520] In another or additional aspect, the present disclosure relates to compositions comprising the fusion proteins / peptides disclosed herein.

[0521] In some embodiments, the composition is used to improve one or more properties of hair, nails, or skin, or a combination thereof, for example, the composition improves the elasticity, moisture retention, or texture of the skin, or provides UV protection to the skin, or a combination of two or more thereof.

[0522] In some embodiments, the composition improves the mechanical properties, color, texture, shape, or moisturization of the hair, or imparts better heat resistance to the hair, or provides a combination of two or more thereof.

[0523] In some embodiments, the composition improves the mechanical or antimicrobial properties of the nail, or provides better UV protection to the nail, or a combination of two or more thereof.

[0524] In some embodiments, the compositions disclosed herein further comprise at least one additive selected from the list of water, preservatives, antioxidants, chelating agents, UV protection agents, vitamins, dyes, hair dyes, surfactants, detergents, emulsifiers, lightening agents, volatile ingredients, propellants, liquid vehicles, carriers, salts, pH adjusters, neutralizing agents, buffers, hair conditioning agents, antistatic agents, anti-friction agents, anti-dandruff agents, natural extracts, moisturizers, fragrances, perfumes, oils, emollients, lubricants, butters, penetrating agents, thickeners, viscosity modifiers, polymers, resins, hair setting agents, film formers, absorbents, and combinations thereof to achieve an approximation of a commercially available product form.

[0525] In some embodiments, the compositions disclosed herein are formulated as soaps, shampoos, conditioners, sprays, gels, low-to-medium viscosity liquids, lotions, milks, mousses, or creams comprising the fusion proteins / peptides of the present disclosure.

[0526] In some embodiments, additives are included or excluded from the composition depending on the form of use.

[0527] Further or additional aspects of the present disclosure relate to methods for obtaining the fusion proteins / peptides disclosed herein.

[0528] In some embodiments, the method for obtaining a fusion protein of the present disclosure comprises: (a) inoculating a liquid culture medium with a microorganism containing an expression gene for a desired fusion protein / peptide; (b) growing the microorganism for a specific cultivation period and inducing expression of the desired fusion protein / peptide under the influence of a promoter; (c) removing endogenous contaminants from the resulting microorganisms to recover the desired fusion protein / peptide; Includes:

[0529] In some embodiments, the method for obtaining the fusion protein / peptide comprises the step of inoculating a liquid culture with a microorganism, in particular an E. coli strain, containing an expression gene for the desired fusion protein / peptide.

[0530] In some embodiments, the method for obtaining the fusion protein comprises recovering the desired fusion protein / peptide from the microorganism by purifying the fusion protein / peptide from endogenous contaminants.

[0531] In some aspects, methods are provided for providing the fusion peptides described herein.

[0532] (definition) Unless otherwise defined, all technical terms, symbols, and other technical and scientific terms or terminology used herein shall have the meanings commonly understood by those skilled in the art. In some cases, commonly understood terms are defined herein for clarity and / or ease of reference, but the provision of such definitions should not necessarily be construed as a substantial difference from what is commonly understood in the field.

[0533] Throughout this disclosure, various embodiments may be presented in a range format. Descriptions in range format are for convenience and conciseness and should not be construed as rigid limitations on the scope of the disclosure. Thus, the description of a range should be considered to specifically disclose all possible subranges and individual numerical values ​​within the range. For example, a description of a range of 1 to 6 should be considered to specifically disclose subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., as well as individual numerical values ​​within the range such as 1, 2, 3, 4, 5, 6, etc. This applies regardless of the width of the range.

[0534] As used herein and in the claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. For example, the term "sample" includes multiple samples and mixtures thereof.

[0535] When the terms "at least," "greater than," or "greater than or equal to" precede the first number of a series of two or more numbers, the terms "at least," "greater than," or "greater than or equal to" apply to each number in the series. For example, 1, 2, or 3 or more is equivalent to 1 or more, 2 or more, or 3 or more.

[0536] The terms "determining," "measuring," "evaluating," "assessing," "assay," and "analysis" are often used interchangeably herein and refer to forms of measurement. These terms include determining whether an element is present or not (e.g., detecting). These terms can include quantitative, qualitative, or quantitative and qualitative determinations. Evaluation can be relative or absolute. "Detecting the presence" can include determining the amount present, in addition to determining presence or absence, depending on the context.

[0537] As used herein, "about" a numerical value refers to ±10% of that numerical value. "About" a range refers to a range of 10% down from the minimum value and 10% up from the maximum value of that range.

[0538] As used herein, the terms "pharmaceutically acceptable" and "cosmetically acceptable" are used interchangeably and refer to compounds, materials, compositions and / or dosage forms that, from a medical standpoint, can be used in contact with human or animal tissue without undue toxicity, irritation, allergic response, or other problem or complication, consistent with a reasonable benefit / risk ratio. More specifically, pharmaceutically acceptable refers to a material, compound, or composition that is suitable for contact with the skin, scalp, or hair. Pharmaceutically acceptable materials are known to those skilled in the art.

[0539] As used herein, the terms "treatment" or "treating" refer to a pharmaceutical or other intervention regimen to obtain a beneficial or desired result in a recipient. Beneficial or desired results include, but are not limited to, therapeutic benefit and / or prophylactic benefit. Therapeutic benefit may refer to the eradication or alleviation of the symptom or underlying disease being treated. Therapeutic benefit may also be achieved when a subject experiences improvement due to the eradication or alleviation of one or more physiological symptoms associated with the underlying disease, although the subject may still be afflicted with the underlying disease. Prophylactic benefit includes delaying, preventing, or eliminating the onset of a disease or condition, delaying or eliminating the onset of symptoms of a disease or condition, slowing, halting, or reversing the progression of a disease or condition, or a combination thereof. For prophylactic benefit, subjects at risk of developing a particular disease or who report one or more physiological symptoms of a disease may receive treatment even if the disease has not been diagnosed.

[0540] As used herein, the terms "hair care" or "hair treatment" refer to compositions for improving the properties of hair, including, but not limited to, strength, elasticity, shine, smoothness, volume, or combinations thereof. "Hair care" or "hair treatment" may also be used to describe the process of applying a composition to hair that has been previously treated or damaged, such as by bleaching or heating.

[0541] When the term "at least," "greater than," or "greater than or equal to" precedes the first number of a series of two or more numbers, the term "at least," "greater than," or "greater than or equal to" applies to each number in the series. For example, 1, 2, or 3 or more is equivalent to 1 or more, 2 or more, or 3 or more.

[0542] When the terms "at most," "less than," or "less than or equal to" precede the first number of a series of two or more numbers, the terms "at most," "less than," or "less than or equal to" apply to each number in the series. For example, 3, 2, or 1 or less is equivalent to 3 or less, 2 or less, or 1 or less.

[0543] The phrases "at least one of A and B" and "at least one of A or B" may be interpreted to mean at least A, at least B, or at least A and B (i.e., a set that comprises A and B, and may include additional elements). The term "A and / or B" may be interpreted to mean A only, B only, or both A and B.

[0544] The phrases "at least about A, B, and C" and "at least about A, B, or C" can be interpreted to mean at least about A, at least about B, or at least about C. The phrases "at most about A, B, and C" and "at most about A, B, or C" can be interpreted to mean at most about A, at most about B, or at most about C.

[0545] The phrase "between about A and B, C and D, and E and F" may be interpreted to mean between about A and about B, between about C and about D, and between about E and about F. The phrase "between about A and B, C and D, or E and F" may be interpreted to mean between about A and about B, between about C and about D, or between about E and about F.

[0546] The phrase "about A to B and C to D" may be interpreted to mean between about A and about B and between about C and about D. The phrase "about A to B or C to D" may be interpreted to mean between about A and about B or between about C and about D.

[0547] The term "comprising" as used in this document indicates the presence of stated features, integers, steps or components and does not exclude the presence or addition of other features, elements, steps, components or groups thereof. Example

[0548] The present disclosure may be further understood by reference to the following examples. The present disclosure is not limited in scope by the embodiments described in the examples, which are intended to be merely illustrative of one aspect of the present disclosure. Any functionally equivalent method is within the scope of the present disclosure. Various modifications of the present disclosure, in addition to those described herein, will be apparent to those skilled in the art from the foregoing description and accompanying drawings. Such modifications are within the scope of the appended claims.

[0549] (Vector construction) The target nucleic acid sequence encoding the fusion protein is inserted into an appropriate expression vector, phage, or directly into the genome or derivative thereof of a microorganism to obtain a polynucleotide composition. The target nucleic acid sequence encodes a fusion protein / peptide of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NOs:44-47, or 200-552, or any combination thereof, or any nucleic acid sequence encoding a fusion protein with at least 60% homology to any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NOs:44-47, or 200-552. The target nucleic acid sequence comprises at least one keratin or keratin-associated peptide component of SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:66, or SEQ ID NO:67, or SEQ ID NOs:70-81, and at least one of an ELP component (SEQ ID NOs:11-23, 49-55, 82-91), an SLP component (SEQ ID NOs:24-32, 56, 92), a CLP component (SEQ ID NOs:33, 57-59, 93-95), an RLP component (SEQ ID NOs:34-35, 60, 96), an ALP component (SEQ ID NOs:36-42, 61), or any combination thereof.

[0550] The expression vector comprises a promoter and a target nucleic acid sequence encoding a target fusion protein / peptide, and optionally comprises a polynucleotide encoding antibiotic resistance (e.g., ampR gene, camR gene, kmR gene, tetR gene, etc.), affinity tag (e.g., C-terminal tag or N-terminal tag), expression reporter, cleavable linker, terminator (e.g., T7 terminator, etc.), ribozyme, or any combination thereof.

[0551] The promoter is included in the vector backbone and is positioned upstream of the start codon. The promoter may be a T7 promoter, lac, T7lac, araBAD, Sp6, trp, or any combination thereof. The promoter may be an inducible or constitutive promoter, which induces overexpression continuously or in the presence of an inducer, respectively. For example, but not limited to, the lac promoter may be incorporated into the vector and induce overexpression in the presence of isopropyl-β-D-thiogalactoside (IPTG).

[0552] The vector comprises an affinity tag, which may be a C-terminal or N-terminal affinity tag. The affinity tag may comprise a polyhistidine tag (3xHis, 4xHis, 5xHis, 6xHis, 7xHis, 8xHis, or 9xHis), a GST tag, a FLAG tag, an MBP tag, a streptavidin (Strep) tag or a derivative of a streptavidin tag (e.g., Strep-tag II, Twin-Strep), a calmodulin tag, a chitin-binding tag, or any combination or derivative thereof. When an N-terminal affinity tag is used, a linker sequence is disposed between the polynucleotide encoding the target fusion protein and the N-terminal affinity tag. When a C-terminal affinity tag is used, a cleavable linker sequence is disposed between the 3' end of the nucleic acid sequence encoding the C-terminal tag and the 5' start codon of the polynucleotide encoding the target fusion protein. The linker sequence encodes a cleavable linker motif that is recognized as a substrate for a protease that selectively cleaves the cleavable linker but not the target fusion protein. For example, a cleavable linker sequence comprising the sequence IEGR is a known substrate for Factor Xa. Introduction of a nucleic acid sequence encoding IEGR-(His)6 allows for expression of the affinity tag 6xHis and the cleavable linker sequence IEGR, with the affinity tag removed after purification. A spacer sequence is optionally positioned between the cleavable linker sequence and the affinity tag sequence.

[0553] Representative vector maps of the expression vectors used are shown in Figures 1 and 2. In Figure 1, a promoter is located upstream of an N-terminal affinity tag sequence, followed by a linker sequence and a target nucleic acid sequence. The expression vector contains an antibiotic resistance gene that confers ampicillin resistance to the host bacterium via the AmpR gene and the AmpR promoter. In Figure 2, a promoter is located upstream of the target nucleic acid sequence, a linker sequence, and a C-terminal affinity tag sequence. The expression vector contains an antibiotic resistance gene that confers ampicillin resistance to the host bacterium via the AmpR gene and the AmpR promoter.

[0554] The target nucleic acid sequence encoding the target fusion protein is inserted into a vector backbone using conventional assembly or Gibson assembly by PCR. In conventional assembly (TA), primers are designed to contain restriction enzyme sites and the target nucleic acid sequence to be inserted into the vector backbone. In Gibson assembly of nucleic acid fragments, Gibson primers are designed to have complementarity to (1) a region of the vector backbone and the nucleic acid sequence encoding the target fusion protein / peptide, or (2) two different regions of the target nucleic acid sequence. Gibson assembly cloning uses the Gibson Assembly® Cloning Kit (New England Biolabs, Inc.).

[0555] (transformation) After the target nucleic acid sequence is cloned into the vector, the insertion of the target nucleic acid sequence is verified by DNA gel (e.g., agarose gel). After confirmation, the vector is transformed into microbial cells via heat shock or electroporation. In one example, the E. coli strain is selected from the group consisting of K-12, B, or their derivatives. In another example, the vector containing the target sequence is introduced into Komagataella phaffii by electroporation, and the gene is integrated into the genome by homologous recombination. The K. phaffii strain is selected from the group consisting of GS115, X-33, or their derivatives.

[0556] (Heat shock transformation) Thaw a stock solution of competent cells on ice and mix with a ligation mixture (e.g., T4 ligase) and an appropriate volume of a vector carrying a polynucleotide sequence encoding the target fusion protein / peptide. Gently mix the mixture and transfer it to a warm water bath or preheated block at approximately 42°C for approximately 30 seconds. Immediately cool the heated mixture on ice and let it sit for approximately 2 minutes. Mix the cooled mixture with LB medium. Incubate the cooled mixture at 37°C for approximately 30-60 minutes before plating. Plate an aliquot of the mixture and incubate it overnight at 37°C.

[0557] (electroporation) Bring the round-bottom culture tube to room temperature. Preheat the recovery medium in a 37°C water bath. Chill the electroporation cuvette on ice. Chill the source of competent E. coli cells on ice and mix with the expression vector containing the target nucleic acid sequence. Dispense a solution containing a 25:1 ratio of competent cells to vector into a microtube and mix gently. Electroporate the microtube containing the cell / DNA mixture in an electroporator (Gene-Pulser, Bio-Rad) at 2.1 kV, 100 Ω, 25 μF, with a time constant of 2.6 ms. Add the warmed recovery medium to the electroporated cell / DNA mixture and shake vigorously at 37°C for approximately 1 hour. Dilute the cells, plate them, and culture them overnight at 37°C.

[0558] (Successful transformation) Successful transformation is determined by taking an aliquot of the bacterial culture (as described below), lysing the culture, extracting the plasmid (e.g., using a DNA extraction kit (QIAamp DNA kit, QIAGEN®)), and comparing the relative molecular weights of the culture-derived plasmid and the empty vector on a DNA gel (e.g., agarose gel). DNA extracted from the culture will have a higher molecular weight than the empty vector. If traditional assembly is used to insert the target nucleic acid sequence, digestion with the corresponding restriction enzyme is performed before running the DNA gel.

[0559] (cell culture) Inoculate colonies into growth medium (e.g., LB). Mix the inoculated media with and without the selection marker. Grow the culture on a shaker at 37°C for several hours. Check the growth stage by taking an aliquot of the culture and measuring the absorbance (or optical density) at 600 nm. The optical density at 600 nm (OD 600 ) corresponds to the exponential or logarithmic growth phase, the culture is removed from the shaker.

[0560] (small scale) Add approximately 25 mL to approximately 100 mL of culture to multiple sterile baffled flasks containing medium and grow overnight on a shaker at 37°C. 600 When OD corresponds to the induction phase, IPTG (or other promoter inducer) is added to the culture, and the OD 600 Shake at 37°C until the cells reach the exponential or logarithmic growth phase. Then, pellet the cells by centrifugation at approximately 1,800 x g for approximately 30 minutes at 4°C. Discard the supernatant and store the pellet at -20°C for later use.

[0561] (large scale) A bioreactor (e.g., BioFlo®, Eppendorf) is equipped with a stirring device (e.g., a baffled stirrer), sterilized, and filled at least halfway with fresh medium. Antifoaming agents are optionally included in the medium. The medium is stirred at about 300 rpm to about 1200 rpm. The bioreactor is maintained at an internal temperature of about 37°C. The stirring rate can be automatically adjusted based on other parameters, such as carbon dioxide (CO2) content. A quorum of transformed cells is slowly introduced into the bioreactor (e.g., via a peristaltic pump). The air composition within the bioreactor is monitored. The air composition, including the amount of CO2, in the headspace within the bioreactor is monitored. The amount of dissolved oxygen (DO) is monitored and maintained at about 35% DO. The pH of the culture medium is also monitored. A pH maintenance feed is fluidly connected to the bioreactor and includes an acidic buffer feed and a basic buffer feed. The pH of the culture medium in the bioreactor is maintained at about 7.2 to about 7.6. A food feed is fluidly connected to the bioreactor and contains glucose (e.g., 20 g / L). The food feed is slowly introduced into the bioreactor. An automated program is used to introduce glucose when the DO content exceeds a predetermined threshold for growth. An aliquot of the culture is taken and the OD 600 Monitor the growth stage of the culture by measuring OD 600 When OD corresponds to the induction phase or late induction phase, an appropriate amount of IPTG or other inducer is added to induce expression. A reporter protein such as green fluorescent protein (GFP) can be expressed, and the green fluorescence color of the culture medium is used to estimate the expression of the target fusion protein. 600 When the growth phase corresponds to the exponential or logarithmic growth phase, the culture is pumped from the bioreactor and pelleted by centrifugation at approximately 1,800 x g for approximately 20 to 30 minutes at 4°C. The supernatant is discarded and the pellet is stored at -30°C.

[0562] (Protein purification) The cell pellet is resuspended in a buffer solution (e.g., PBS buffer, pH 7.4). An appropriate amount of buffer is added to the pellet to form a suspension. The pellet and buffer solution are mixed until homogenous. Using a vortexer or homogenizer, the suspension is lysed by passing through a French press, sonication, high-pressure homogenization, mixing with glass beads, or multiple freeze / thaw cycles. Protease inhibitors are added to the lysed cell suspension. The lysed cells are centrifuged at least 2,000 x g at 4°C for approximately 5 to 30 minutes. The supernatant is collected and the pellet is discarded. The supernatant is purified by size exclusion chromatography, affinity chromatography, or direct drying. The pure fraction is then dialyzed or diafiltered against buffer or water.

[0563] (Chromatography) (size exclusion) The concentrated supernatant is passed through a size-exclusion column. The column is selected based on the desired molecular weight resolution. For example, but not limited to, if the fusion protein has a molecular weight of approximately 35 kDa, a size-exclusion resin with a resolution corresponding to proteins of 5 kDa to 250 kDa (e.g., HiPrep Sephacryl S-200 HR, Cytiva LifeSciences) is used.

[0564] (affinity chromatography) The affinity chromatography medium is selected based on the affinity tag incorporated into the expressed fusion protein. When the 6xHis tag is incorporated into the expression vector, the expressed 6xHis tag is suitable for affinity chromatography with Ni(II)-NTA resin. An HPLC instrument (AKTA Pure Protein Purification System, Cytiva LifeSciences) equipped with Buffer A and Buffer B is used. Buffer A and Buffer B are fluidly connected to the affinity chromatography medium, e.g., a Ni(II)-NTA column. Buffer A contains 0 mM imidazole. Buffer B contains approximately 200 mM imidazole. The column is primed with several column volumes of Buffer A, followed by several volumes of Buffer B diluted in Buffer A. The concentrated supernatant is loaded onto the column and allowed to incubate for several minutes. Unwanted components are eluted using a ramp from 0 mM imidazole to approximately 150 mM imidazole. The purified target fusion protein is eluted using a final wash of several column volumes of 100% Buffer B (200 mM imidazole). The purified target fusion protein / peptide is collected for further processing.

[0565] (Further purification - cleavage of affinity tag) The purified His-tagged target fusion protein / peptide is further treated with a protease that selectively cleaves the cleavable linker to yield a purified, untagged target fusion protein / peptide. The untagged target fusion protein / peptide is then passed through a spin column with an appropriate molecular weight cutoff and concentrated using a tabletop centrifuge.

[0566] (characteristic evaluation) The purified and concentrated fusion protein / peptide can be characterized by SDS-PAGE, circular dichroism (CD) spectroscopy, nuclear magnetic resonance (NMR), mass spectrometry (MS), etc.

[0567] (SDS-PAGE) A protein gel (e.g., SDS-PAGE) is run with serial dilutions of the concentrated target fusion protein / peptide against a ladder. The protein gel shows a band corresponding to the desired molecular weight of the target fusion protein / peptide excluding the cleavable linker motif and affinity tag. The concentrated target fusion protein / peptide is pre-digested with an enzyme to obtain digestion products, which are then evaluated by SDS-PAGE.

[0568] (Circular dichroism spectroscopy) The secondary structure of the concentrated target fusion protein / peptide was analyzed by circular dichroism spectroscopy (CD; J-1000 Series Circular Dichroism Spectropolarimeter, Jasco). A maximum peak was observed at 195 nm, and minimum peaks at 208 and 222 nm, corresponding to the α-helix of the target fusion protein / peptide. A maximum peak was observed at 195 nm, and minimum peaks were observed at 218 nm, corresponding to the β-sheet of the target fusion protein. A minimum peak was observed at 198 nm, corresponding to the random coil of the target fusion protein.

[0569] (mass spectrometry) The enriched target fusion proteins / peptides can be analyzed by mass spectrometry (MS), e.g., liquid chromatography-mass spectrometry (LC-MS 2 The enriched target fusion proteins / peptides are pretreated with DTT and optionally digested in the presence of proteases (e.g., chymotrypsin, trypsin, LysC, AspN). The enriched target fusion proteins / peptides are mixed with 1.0% by volume trifluoroacetic acid (TFA) before fragmentation and then fragmented by ESI-MS (Orbitrap, ThermoFisher). Data are collected and processed using Proteome Discoverer (ThermoFisher). Expected fragment sizes for various fragmentation and charge states are estimated using Proteome Discoverer or ExPasy.

[0570] The fusion protein / peptide (0.01%) solution was applied in an ethanol formulation to bleached Asian hair tresses (300 mg). Several properties were evaluated, including emulsifying properties, antimicrobial properties, fiber integrity, elasticity, Young's modulus, hydrophobicity, and others.

[0571] (Emulsifier properties) The emulsifier properties of the peptides were evaluated by adding each peptide at a concentration of 0.1% (m / v) to a mixture containing 15% edible oil and water. The pH of the samples was adjusted to 7. To promote emulsification, the samples were sonicated for 20 seconds. The samples were observed immediately after sonication and 7 days later. The evaluation was classified as "yes" if the peptide exhibited emulsifier properties and "no" if it did not. The results of the study are shown in Table 3.

[0572] [Table 3]

[0573] (Antibacterial - Pharmaceutical or Cosmetic) Antimicrobial susceptibility testing of peptides was performed according to CLSI and EUCAST recommendations. Assays were prepared in lysogeny medium. Pre-cultures of Pseudomonas aeruginosa and Staphylococcus aureus were prepared in advance, and a final concentration of 1 × 10 was determined by a pre-established calibration curve. 6 The concentrations were adjusted to CFU / mL. Minimum inhibitory concentration assays were prepared in 96-well plates over a concentration range of 0.78–400 μM. The peptides were scored as “yes” if they showed antibacterial activity against both bacterial species, and “no” if they showed no antibacterial activity against either bacterial species. The results of this study are shown in Table 4.

[0574] [Table 4]

[0575] (fiber integrity) The fiber integrity of hair samples with peptides (2 ± 0.1 mg) was analyzed by differential scanning calorimetry (DSC) using a Netzsch DSC 3500 Sirius. Thermal studies of the effects of selected peptides on hair were performed using a power-compensated differential scanning calorimeter and an aluminum pan (maximum pressure: 1 bar) over a temperature range of 25 °C to 150 °C (heating rate: 10 °C / min), followed by an isothermal hold at 150 °C for 5 min, followed by a heating rate of 10 °C / min to 300 °C. Data are presented as the mean ± standard error of replicate measurements, and hair integrity values ​​are expressed as a percentage compared to water-treated hair (enthalpy = 16.4 ± 0.6). The results of this study are shown in Table 5.

[0576] [Table 5]

[0577] (Young's modulus) The mechanical properties (Young's modulus) of hair fibers were evaluated using a texture analyzer (TA.XTplusC, Stable Micro Systems) to study the effects of selected peptide treatments. Differences in hair mechanical properties were determined for 30 single hair fibers with small diameter variations. Fibers were individually mounted on a tensile fixture (a paper template with a fixed gauge length of 55 mm). Test samples were stored in an exciter to avoid excessive humidity. Elongation measurements were performed at a speed of 0.03 mm / s. This parameter was evaluated as the percentage of hair recovery when comparing peptide-treated hair with damaged, bleached hair (untreated). Data are presented as mean ± standard error. The results of this study are shown in Table 6.

[0578] [Table 6]

[0579] (hydrophobic properties) Advancing contact angles were measured using a Force Tensiometer K100C (KRUSS). Contact angles in water were calculated using the force and estimated fiber diameter. During the measurements, 10 individual hair fibers were immersed to a predetermined depth at a rate of 5 mm / min. The results of this study are shown in Table 7.

[0580] [Table 7]

[0581] The penetration / localization of the peptides within the hair fiber was observed by labeling the peptides with FITC (a fluorescent probe) and observing them under a fluorescent microscope (Figure 3).

[0582] (Computer simulation of fusion protein interaction with model keratin) The complexation free energy of the fusion peptide and the model keratin was modeled. The free energy was calculated, and the number of hydrogen bonds is shown in Table 8.

[0583] [Table 8]

[0584] A more negative ΔG corresponds to a stronger interaction between the peptide and the keratin model. The number of hydrogen bonds between the peptide and the keratin model is also a direct indicator of their relationship. These parameters indicate how the fusion peptide interacts with keratin.

[0585] Alignment methods for sequence comparison are widely known to those skilled in the art and include GAP, BESTFIT, BLAST, FASTA, TFASTA, etc. GAP uses the algorithm of Needleman and Wunsch ((1970) J Mol Biol, 48:443-453) to perform a global alignment across two sequences, maximizing the number of matches and minimizing the number of gaps. The BLAST algorithm (Altschul et al. (1990) J Mol Biol 215:403-10) calculates the percent identity between sequences and performs statistical analysis of the similarity between two sequences. Software for BLAST analysis is publicly available through the National Center for Biotechnology Information (NCBI). Global similarity and identity percentages can also be determined using any of the methods available in the MatGAT software package (Campanella et al., BMC Bioinformatics, 2003, July 10; 4:29. MatGAT: an application for generating similarity / identity matrices using protein or DNA sequences). As will be apparent to one skilled in the art, minor manual editing can be performed to optimize alignment between conserved motifs. The percentage sequence identity values ​​presented in the presently disclosed subject matter were determined across the entire amino acid sequence using the default parameters of BLAST.

[0586] In the claims herein, where the singular form of an element or feature is used, the plural form is also included, and vice versa, unless specifically excluded. For example, the term "a peptide" or "the peptide" includes the plural form "peptides" or "the peptides," and vice versa. In the claims, articles such as "a," "an," and "the" can mean one or more, unless specifically stated to the contrary or clearly evident from the context. Any claim or description containing "or" between one or more members of a group is considered to be satisfied when one, more than one, or all of the members are present in, used in, or otherwise relevant to a product or process. The disclosure includes embodiments in which exactly one member of a group is present in, used in, or otherwise relevant. It also includes embodiments in which more than one or all of the members are present in, used in, or otherwise relevant.

[0587] Furthermore, the disclosure encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, descriptive terms, etc. are introduced into one or more claims or relevant portions of the specification from other claims. For example, a claim that is dependent on another claim may be modified to include one or more limitations set forth in the other claims that are dependent on the same base claim.

[0588] Furthermore, when a claim recites a composition, it is to be understood that the claim also encompasses methods of using the composition for any purpose disclosed herein, and methods of making the composition by any method disclosed herein or other methods known to those of skill in the art, unless specifically stated to the contrary or unless one of skill in the art would determine that a contradiction or inconsistency would arise.

[0589] When ranges are expressed, the endpoints are included. Furthermore, unless otherwise clearly indicated by description, context, or the understanding of one of ordinary skill in the art, values ​​expressed in ranges should be understood to be able to take any specific value, in different embodiments of the disclosure, up to one digit of the unit of the lower limit of the range. Furthermore, unless otherwise clearly indicated by description, context, or the understanding of one of ordinary skill in the art, values ​​expressed in ranges should be understood to be able to take any subrange within the range, the endpoints of which are expressed with precision to one digit of the unit of the lower limit of the range.

[0590] The present disclosure is not intended to be limited in any way to the described embodiments, as those skilled in the art will recognize many possible modifications.

[0591] The above embodiments can be combined.

[0592] The accompanying claims further define particular embodiments of the present disclosure.

Claims

1. A fusion protein / peptide comprising: (a) a keratin or keratin-related peptide block; (b) at least one polypeptide block selected from the list consisting of: (i) an elastin or elastin-like polypeptide (ELP) block, (ii) a silk or silk-like polypeptide (SLP) block, (iii) a collagen or collagen-like polypeptide (CLP) block, (iv) a resilin or resilin-like polypeptide (RLP) block, (v) an abductin or abductin-like polypeptide (ALP) block, or (vi) a combination of two or more blocks selected from (i)-(v).

2. The fusion protein / peptide of claim 1 , wherein the polypeptide block comprises an elastin or elastin-like polypeptide block.

3. 10. The fusion protein / peptide according to any one of the preceding claims, wherein the elastin or elastin-like polypeptide (ELP) block has a length of 3 to 30 amino acids, preferably 5 to 20 amino acids, more preferably 15 to 30 amino acids.

4. 3. A fusion protein / peptide according to claim 1 or claim 2, which comprises a linker block as a spacer.

5. A fusion protein / peptide according to any one of the preceding claims, wherein the linker block has a repeat sequence of 2 to 8 amino acids in length.

6. The fusion protein / peptide according to claim 4 or claim 5, wherein the linker block comprises a repeating sequence having a length of 2 to 40 amino acids, preferably 8 to 30 amino acids.

7. The fusion protein / peptide according to any one of claims 4 to 6, wherein the linker block comprises a hydrophilic block having a length of 1 to 75 amino acids.

8. A fusion protein / peptide according to the preceding claims, wherein said hydrophilic block has a length of 10 to 50 amino acids.

9. 10. A fusion protein / peptide according to any one of the preceding claims, wherein the linker block adopts an alpha helix structure.

10. A fusion protein / peptide according to any one of the preceding claims, wherein said linker has a length of 1 to 3 amino acids which is repeated 0 to 25 times, preferably 1 to 20 times.

11. A fusion protein / peptide according to the preceding claims, wherein said linker block comprises 1 or 2 amino acids repeated 10 to 25 times.

12. The fusion protein / peptide according to any one of the preceding claims, wherein the keratin or keratin-associated peptide block comprises SEQ ID NO: 8:GGVCGPSPPCITT or a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO:

8.

13. The fusion protein / peptide according to any one of the preceding claims, wherein the keratin or keratin-associated peptide block comprises SEQ ID NO: 8:GGVCGPSPPCITT or a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO:

8.

14. The fusion protein / peptide according to any one of the preceding claims, wherein the keratin or keratin-associated peptide block comprises SEQ ID NO: 9:GGVCGPSPPC or a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO:

9.

15. The fusion protein / peptide according to any one of the preceding claims, wherein the keratin or keratin-associated peptide block comprises SEQ ID NO: 10:CGPSPPCITT or a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO:

10.

16. The fusion protein / peptide according to any one of the preceding claims, wherein the keratin or keratin-associated peptide block comprises SEQ ID NO: 68:CLPCLPAASC or a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO:

68.

17. The fusion protein / peptide according to any one of the preceding claims, wherein the keratin or keratin-associated peptide block comprises SEQ ID NO: 73:CVSSCCKPQCC or a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO:

73.

18. The fusion protein / peptide according to any one of the preceding claims, wherein the keratin or keratin-associated peptide block comprises SEQ ID NO: 74:PITCRRTCYH or a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO:

74.

19. The fusion protein / peptide according to any one of the preceding claims, wherein the keratin or keratin-associated peptide block comprises SEQ ID NO: 75:DCKLPCNPCA or a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO:

75.

20. The keratin or keratin-associated peptide block comprises SEQ ID NO: 76:CLPCLPAASC or a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO:

76. Fusion protein / peptide according to any one of the preceding claims.

21. The fusion protein / peptide according to any one of the preceding claims, wherein the keratin or keratin-associated peptide block comprises SEQ ID NO: 77: CEPAICEPSC or a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO:

77.

22. 10. The fusion protein / peptide of any one of the preceding claims, wherein the fusion protein / peptide has SEQ ID NO. 1:GGVCGPSPPCITTVPGVGVPGVGVPGVGLGVPGVGVPGVGVPGVGVGR, or at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO:

1.

23. 10. The fusion protein / peptide of any one of the preceding claims, wherein the fusion protein / peptide is SEQ ID NO. 2:GGVCGPSPPCITTVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVGVG, or has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO:

2.

24. 10. The fusion protein / peptide of any one of the preceding claims, wherein the fusion protein / peptide has SEQ ID NO. 7:GGVCGPSPPCITTVPGVGVPGVGVGVPGLGVPGVGVPGVGVGVG, or at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO:

7.

25. 10. The fusion protein / peptide of any one of the preceding claims, wherein the polypeptide block comprises a silk or silk-like polypeptide (SLP) block.

26. 10. The fusion protein / peptide of any one of the preceding claims, wherein the fusion protein / peptide has SEQ ID NO:3:GGVCGPSPPCITTGAGAGSGAGAGSGAGAGSGAGAGSGAGAS, or at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO:

3.

27. 10. The fusion protein / peptide of any one of the preceding claims, wherein the polypeptide block comprises a collagen or collagen-like polypeptide (CLP) block.

28. 10. The fusion protein of any one of the preceding claims, wherein the fusion protein has SEQ ID NO:5:GGVCGPSPPCITTGPTGPTGPAGPRGLQGLQGLQGERGEQGPT, or at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO:

5.

29. 10. The fusion protein / peptide of any one of the preceding claims, wherein the polypeptide component comprises a resilin or resilin-like polypeptide (RLP) component.

30. 10. The fusion protein of any one of the preceding claims, wherein the fusion protein has SEQ ID NO:4:GGVCGPSPPCITTGGRPSSDSYGAPGGGN, or at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO:

4.

31. 10. A fusion protein / peptide according to any one of the preceding claims, wherein the polypeptide component comprises an abductin or abductin polypeptide (ALP) component.

32. 10. The fusion protein of any one of the preceding claims, wherein the fusion protein has SEQ ID NO. 6:GGVCGPSPPCITTGGFGGMGGGSGGFGGMGGGSGGFGGMGGGGS, or at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO:

6.

33. the keratin or keratin-associated peptide component has a length of 6 to 16 amino acids, and / or The silk or "SLP" has a repeat sequence having a length of 6-8 amino acids, and / or the silk or SLP has a length of 10-30 amino acids, preferably 15-20 amino acids, and / or The collagen or "CLP" has a repeat sequence with a length of 3 amino acids, and / or the collagen or CLP has a length of 10 to 32 amino acids, preferably 15 to 30 amino acids, and / or The elastin or "ELP" has a repeat sequence having a length of 5 amino acids, and / or the elastin or ELP has a length of 3 to 30 amino acids, preferably 8 to 32, more preferably 15 to 30, and / or Resilin or "RLP" has a repeat sequence having a length of 11-15 amino acids, and / or Resilin or RLP has a length of 10-30 amino acids, preferably 15-20 amino acids, and / or 10. The fusion protein / peptide according to any one of the preceding claims, wherein the abductin or "ALP" has a repeat sequence with a length of 4 to 10 amino acids and / or the abductin or ALP has a length of 15 to 32, more preferably 20 to 30.

34. 10. The fusion protein / peptide of any one of the preceding claims, wherein the fusion protein has an average molecular weight of about 1 kDa to about 144 kDa, about 10 kDa to about 144 kDa, or about 35 kDa to about 130 kDa.

35. The fusion protein may be from about 1 kDa to about 5 kDa, from about 5 kDa to about 10 kDa, from about 6 kDa to about 17 kDa, from about 10 kDa to about 15 kDa, from about 14 kDa to about 30 kDa, from about 15 kDa to about 20 kDa, from about 17 kDa to about 39 kDa, from about 20 kDa to about 25 kDa, from about 25 kDa to about 30 kDa, from about 30 kDa to about 35 kDa, from about 35 kDa to about 40 kDa.

2. The fusion protein / peptide according to any one of the preceding claims, having an average molecular weight range of about 39 kDa to about 54 kDa, about 39 kDa to about 80 kDa, about 40 kDa to about 45 kDa, about 45 kDa to about 50 kDa, about 50 kDa to about 55 kDa, about 55 kDa to about 60 kDa, about 60 kDa to about 100 kDa, about 80 kDa to about 144 kDa.

36. 10. The fusion protein / peptide of any one of the preceding claims, wherein the fusion protein / peptide comprises a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 1-7, 43-47, or 200-552.

37. 10. The fusion protein / peptide of any one of the preceding claims, wherein the fusion protein / peptide comprises a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID NOs: 1-7, 43-47, or 200-552.

38. 10. The fusion protein / peptide of any one of the preceding claims, wherein the fusion protein / peptide comprises a sequence identical to any one of SEQ ID NOs: 1-7, 43-47, or 200-552.

39. 10. The fusion protein / peptide according to any one of the preceding claims, wherein the cosmetic product is suitable for hair care or hair treatment.

40. The fusion protein / peptide comprises: [B ｎ１ -A ｍ -B ｎ -C ｏ -B ｎ２ ] ｐ Formula (I), or [B ｎ１ -C ｏ -B ｎ -A ｍ -B ｎ２ ] ｐ Formula (II), or [B ｎ１ -B ｎ -C ｏ -A ｍ -B ｎ２ ] ｐ Formula (III), or [B ｎ１ -C ｏ -A ｍ -B ｎ -B ｎ２ ] ｐ Formula (IV), Represented by formulas (I) to (IV): "A" comprises a peptide sequence comprising said keratin or keratin-associated peptide block, and "m" is an integer from 1 to about 10; "B" comprises a peptide sequence comprising 1 to 3 amino acids, each “n”, “n” １ " and "n ２ " are independently an integer from 0 to 25, "C" is D ｑ -E ｒ -F ｓ -G ｔ -H ｕ a peptide sequence having the structure "D" comprises a silk or silk-like polypeptide (SLP) block; "E" comprises a collagen or collagen-like polypeptide (CLP) block; "F" comprises an elastin or elastin-like polypeptide (ELP) block; "G" comprises a resilin or resilin-like polypeptide (RLP) block; "H" comprises an abductin or abductin-like polypeptide (ALP) block; "q" is an integer from 0 to about 100, "r" is an integer from 0 to about 200, "s" is an integer from 0 to about 220, "t" is an integer from 0 to about 80, and "u" is an integer from 0 to about 70; "o" is an integer from 0 to about 30, and "p" is an integer from about 1 to about 40; "n", "n １ "," "n ２ 2. The fusion protein / peptide of claim 1, wherein at least one of "n", "n", or "o" is not 0. １ " and "n ２ 34. The fusion protein / peptide of claim 33, wherein each of the following is independently an integer from 0 to about 25:

41. The fusion protein / peptide of claim 33 or 34, wherein "o" is an integer from about 1 to about 30.

42. each “n”, “n” １ " and "n ２ 36. The fusion protein / peptide of any one of claims 33 to 35, wherein " is an integer between 0 and 25 and "o" is an integer between 1 and 30.

43. 37. The fusion protein / peptide of any one of claims 33 to 36, wherein "p" is an integer between 3 and 37, between 8 and 15, or between 10 and 12.

44. 38. The fusion protein / peptide of any one of claims 33 to 37, wherein "B" comprises a peptide sequence comprising 1 to 40 amino acids, 2 to 25 amino acids, or 4 to 25 amino acids. "B" is X ｎ Z ｃ and / or Z ｃ X ｎ and a peptide sequence having the formula "X ｎ " is proline, leucine, isoleucine, glutamic acid, aspartic acid, glutamine, lysine, alanine, arginine or threonine, and "Z ｃ 39. The fusion protein / peptide of any one of claims 33 to 38, wherein " comprises glycine, alanine, lysine, proline, methionine, glutamine, arginine, threonine, valine, glutamic acid, aspartic acid or asparagine. said at least one keratin or keratin-associated peptide block having a length of 6 to 16 amino acids; "D" comprises a polypeptide sequence comprising SEQ ID NO:24:GAGAGS, SEQ ID NO:25:GAGSGA, SEQ ID NO:26:GAGAGY, SEQ ID NO:27:GAGYGA, SEQ ID NO:28:GAGAGA, SEQ ID NO:29:GAGAGV, SEQ ID NO:30:GAGVGA, or SEQ ID NO:31:GAGAGVGY, or a combination of two or more thereof; "E" comprises the polypeptide sequence GX1Z, wherein "X1" is proline, leucine, isoleucine, glutamic acid, aspartic acid, glutamine, lysine, alanine, arginine, or threonine; Z is glycine, alanine, lysine, proline, methionine, glutamine, arginine, threonine, valine, glutamic acid, aspartic acid, or asparagine; "F" comprises a polypeptide sequence comprising SEQ ID NO:21:VPGX2G and / or SEQ ID NO:13:VPAVG; "X2" is arginine, histidine, lysine, aspartic acid, glutamic acid, serine, threonine, asparagine, glutamine, cysteine, glycine, alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, or tryptophan; "G" comprises a polypeptide sequence comprising SEQ ID NO: 34: GGRPSDSYGAPGGGGN and / or SEQ ID NO: 35: GAPAQTPSSQY; 39. The fusion protein / peptide of any one of claims 32 to 38, wherein "H" comprises a polypeptide sequence comprising SEQ ID NO: 36: GGFGGMGGGGS, SEQ ID NO: 37: MGGG, SEQ ID NO: 38: FGGMG, SEQ ID NO: 39: FGGMGGG, SEQ ID NO: 40: GGFGGMGGG, or SEQ ID NO: 41: FGGMGGGGNAG, or a combination of two or more thereof.

45. "A" is the peptide sequence X ３ ｋ CX ３ ５ CX ３ ｋ Equipped with "X ３ " is glycine, alanine, valine, isoleucine, leucine, phenylalanine, proline, serine, threonine, asparagine, glutamine, cysteine, methionine, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, histidine, lysine or arginine; 40. The fusion protein / peptide of any one of claims 32 to 39, wherein "k" is an integer from 0 to 3.

46. 41. The fusion protein / peptide of any one of claims 32 to 40, wherein "A" comprises SEQ ID NO: 8: GGVCGPSPPCITT, SEQ ID NO: 9: GGVCGPSPPC, SEQ ID NO: 10: CGPSPPCITT, CGPSPPC (SEQ ID NO: 66), or GVCGPSPPC (SEQ ID NO: 67).

47. 42. The fusion protein / peptide of any one of claims 32 to 41, wherein the fusion protein has an average molecular weight range of about 1 kDa to about 144 kDa, about 10 kDa to about 144 kDa, or about 35 kDa to about 130 kDa.

48. The fusion protein may be from about 1 kDa to about 5 kDa, from about 5 kDa to about 10 kDa, from about 6 kDa to about 17 kDa, from about 10 kDa to about 15 kDa, from about 14 kDa to about 30 kDa, from about 15 kDa to about 20 kDa, from about 17 kDa to about 39 kDa, from about 20 kDa to about 25 kDa, from about 25 kDa to about 30 kDa, from about 30 kDa to about 35 kDa, or from about 35 kDa to about 40 kDa.

43. The fusion protein / peptide of any one of claims 32 to 42, having an average molecular weight range of about 39 kDa to about 54 kDa, about 39 kDa to about 80 kDa, about 40 kDa to about 45 kDa, about 45 kDa to about 50 kDa, about 50 kDa to about 55 kDa, about 55 kDa to about 60 kDa, about 60 kDa to about 100 kDa, or about 80 kDa to about 144 kDa.

49. The fusion protein / peptide of any one of claims 32 to 43, wherein the silk or silk-like peptide (SLP) block forms a beta-sheet rich structure.

50. The fusion protein / peptide of any one of claims 32 to 44, wherein the elastin or elastin-like peptide (ELP) block forms a β-turn rich structure.

51. The fusion protein / peptide of any one of claims 32 to 45, wherein the elastin-like peptide (ELP) block is temperature responsive.

52. 44. A composition comprising the fusion protein / peptide of any one of claims 1 to 43 for improving the elasticity, recovery, moisturization, integrity, hydrophobicity, texture, antibacterial properties, emulsifier properties, mechanical properties, heat resistance or UV protection of hair, nails or skin, or a combination of two or more thereof.

53. 10. The composition of the preceding claim, wherein the composition improves skin elasticity, recovery, moisturization, integrity, texture, antimicrobial or UV protection, or a combination of two or more thereof.

54. 46. ​​The composition of any one of claims 44 to 45, wherein the amount of the fusion protein / peptide is in the range of 0.0001% to 20% (w / w), preferably in the range of 0.001% to 5% (w / w), or 0.1% to about 1% (w / w).

55. 47. The composition of any one of claims 43 to 46, wherein the composition improves the mechanical properties, color, texture, shape, moisturization, integrity, recovery, hydrophobicity or heat resistance of hair, or a combination of two or more thereof.

56. 48. The composition of any one of claims 44 to 47, wherein the composition improves the mechanical properties, integrity, antibacterial properties, or UV protection of the nail, or a combination of two or more thereof.

57. 49. The composition of any one of claims 44 to 48, wherein the at least one excipient suitable for use on the skin is selected from surfactants, emulsifiers, preservatives, thickeners, organic polymers, moisturizers, silicones, oils, fragrances, vitamins, buffers, antimicrobial agents, antibacterial agents, disinfectants, chelating agents or mixtures thereof.

58. 50. The composition of any one of claims 44 to 49, comprising at least one excipient suitable for use on the skin.

59. 10. The composition of claim 9, wherein the at least one excipient suitable for skin application is selected from ethanol, benzyl alcohol, a diol molecule, urea, ammonium thioglycolate, thioanisole, tris(hydroxymethyl)aminomethane, phosphate buffer, sodium chloride, citrate buffer, or ethanol, or a combination of two or more thereof.

60. 10. The composition of any one of the preceding claims, wherein the diol molecule is selected from ethylene glycol, propylene glycol, butylene glycol, butylene diol or mixtures thereof.

61. 0 to 30% (w / v), preferably 10 to 20% (w / v) ethanol; 0-3% (w / v), preferably 0.5-2% (w / v) benzyl alcohol; 0-20% (w / v), preferably 10-15% (w / v) of diol molecules, Including, 53. The composition according to any one of claims 44 to 52.

62. 54. The composition of any one of claims 44 to 53, further comprising a propellant, a fragrance, an oil, or a mixture thereof.

63. A composition according to any one of claims 44 to 54 for use in treating hair.

64. 10. A fusion protein / peptide or composition according to any one of the preceding claims for use as a cosmetic, i.e. as a hair, nail or skin cosmetic.

65. The fusion protein / peptide or composition according to any one of claims 44 to 56, wherein the cosmetic product is suitable for hair care or hair treatment.

66. 10. A shampoo, lotion, serum, cream, conditioner, foam, elixir, oil, aerosol or mask comprising a composition / fusion protein / peptide according to any one of the preceding claims.

67. 44. Use of a composition comprising a fusion protein / peptide according to any one of claims 1 to 43 as an agent for enhancing one or more properties of hair, nails or skin, or as an agent for enhancing elasticity, moisture retention, recovery, texture, antibacterial properties and / or sun protection of hair, nails and skin.

68. inoculating a liquid culture medium with a microorganism containing an expression gene for a desired fusion protein; allowing the microorganism to grow for a specific culture period and inducing expression under the influence of a promoter; recovering the desired fusion protein / peptide from the obtained microorganism by purifying the protein from endogenous contaminants; A method for obtaining a fusion protein / peptide according to any one of claims 1 to 43, comprising:

69. 10. The method of the preceding claim, wherein the microorganism is Escherichia coli.

70. 62. The method of any one of claims 60 to 61, wherein the fusion protein / peptide is purified from endogenous contaminants.

71. Initiating solid-phase synthesis by combining a keratin or keratin-related peptide block with at least one polypeptide block selected from the list consisting of: (i) an elastin or elastin-like polypeptide (ELP) block, (ii) a silk or silk-like polypeptide (SLP) block, (iii) a collagen or collagen-like polypeptide (CLP) block, (iv) a resilin or resilin-like polypeptide (RLP) block, (v) an abductin or abductin-like polypeptide (ALP) block, or (vi) a combination of two or more blocks selected from (i) to (v); performing multiple solid-phase reactions to obtain said fusion protein / peptide; A method for obtaining a fusion protein / peptide according to any one of claims 1 to 51 by solid phase synthesis comprising:

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