Biological hair shape modifying compositions and kits, and methods for altering hair shape

Abstract

To provide novel biological perm agents that can reduce the risk of skin redness, allergies, and the like and have an excellent shape-imparting effect.SOLUTION: A biological hair modification composition is provided that contains a macromolecular component, a small molecule component and an alkaline agent component. The macromolecular component contains a protease belonging to alkaline proteases, and the small molecule component contains a peptide with reducing activity. Both macromolecular and small molecule components are obtained from the fermentation product of Bacillus licheniformis and a keratin- and / or keratin polymer-containing medium. The molecular weight of the components in the macromolecular component is greater than or equal to 3 kDa and is between 3 and 1000 kDa, and the molecular weight of the components in the small molecule component is less than 3 kDa and is between 0.01 and 2.99 kDa.SELECTED DRAWING: None

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application is based on and claims priority to Taiwan Patent Application No. 110126224, filed on July 16, 2021, the disclosure of which is incorporated by reference in its entirety herein.

[0002] The technical field of this disclosure relates to biological hair shape modifying compositions and kits, and methods for altering hair shape. [Background technology]

[0003] The commonly used perm method is achieved by changing the disulfide bonds in the keratin of the hair with a perm agent so that the hair can be reshaped. The perming steps mainly include breaking the disulfide bonds in the keratin of the hair with a reducing agent, reshaping the hair with a styling tool, and rearranging and forming the disulfide bonds with an oxidizing agent, thereby making the hair take the reshaped shape.

[0004] Currently used chemical perm agents are effective, but they are prone to cause scalp redness, skin irritation, skin allergies, and hair damage. In addition, commercially available chemical perm agents have the risk of inducing cancer. Currently, it is known that amino acid agents containing small amounts of chemical components are available for perm treatment, but they are less damaging to disulfide bonds, so the effect of hair styling is low. Summary of the Invention [Problem to be solved by the invention]

[0005] Therefore, there is an urgent need for a novel biological perm agent that can reduce the risks of skin redness, allergies, etc., and at the same time has an excellent shaping effect.

[0006] The present disclosure provides a biological hair shape modification composition comprising a macromolecular component, a small molecular component, and an alkaline agent component. The macromolecular component comprises a protease belonging to the alkaline protease family, and the small molecular component comprises a peptide having reducing activity. Both the macromolecular component and the small molecular component are obtained from a fermentation product of Bacillus licheniformis and a medium containing keratin and / or a keratin polymer. The molecular weight of the component in the macromolecular component is 3 kDa or more and 3 to 1000 kDa, and the molecular weight of the component in the small molecular component is less than 3 kDa and 0.01 to 2.99 kDa.

[0007] The present disclosure also provides a biological hair shape modification kit including a polymer component, a small molecule component, an alkaline agent component, and at least one packaging container for containing the polymer component, the small molecule component, and the alkaline agent component. The polymer component includes a protease belonging to an alkaline protease family, and the small molecule component includes a peptide having reducing activity. Both the polymer component and the small molecule component are obtained from a fermentation product of Bacillus licheniformis and a medium containing keratin and / or a keratin polymer. The molecular weight of the component in the polymer component is 3 kDa or more and 3 to 1000 kDa, and the molecular weight of the component in the small molecule component is less than 3 kDa and 0.01 to 2.99 kDa. The biological hair shape modification kit is used in the form of a mixture of the polymer component, the small molecule component, and the alkaline agent component.

[0008] The present disclosure further provides a method for changing hair shape, comprising: (a) a hair permeation step of applying the above-mentioned biological hair shape modification composition or a mixture prepared using the above-mentioned biological hair shape modification kit to hair and permeating it into the hair; and (b) a hair shape modification step of subjecting the hair to which the above-mentioned biological hair shape modification composition or the above-mentioned mixture has been applied and permeated, to a heating step to hydrolyze the backbone of the hair protein and cleave disulfide bonds in the hair protein, thereby changing the shape of the hair.

[0009] This specification includes the disclosure of the following inventions. [1] A polymer component containing a protease belonging to the alkaline protease class; a small molecule component comprising a peptide having reducing activity; An alkaline agent component; A biological hair shape modifying composition comprising: Both the macromolecular component and the small molecule component are obtained from the fermentation product of Bacillus licheniformis and a medium containing keratin and / or a keratin polymer; and The molecular weight of the component in the polymer component is 3 kDa or more and 3 to 1000 kDa; A biological hair shape modifying composition, wherein the molecular weight of the component in the small molecule component is less than 3 kDa and is 0.01 to 2.99 kDa. [2] The biological hair shape altering composition according to [1], wherein the protease belonging to the alkaline protease class includes alkaline protease, keratinase, or a combination thereof. [3] The biological hair shape modifying composition according to [1] or [2], wherein the peptide having reducing activity includes at least one of the following peptides: (a) a peptide comprising the amino acid sequence SEQ ID NO.1: (b) a peptide comprising the amino acid sequence SEQ ID NO.2; (c) a peptide comprising the amino acid sequence SEQ ID NO.3; (d) a peptide comprising the amino acid sequence SEQ ID NO.4; (e) a peptide comprising the amino acid sequence SEQ ID NO.5; (f) a peptide comprising the amino acid sequence SEQ ID NO.6; (g) a peptide comprising the amino acid sequence SEQ ID NO.7; (h) a peptide comprising the amino acid sequence SEQ ID NO.8; (i) a peptide comprising the amino acid sequence SEQ ID NO.9; and (j) a peptide comprising the amino acid sequence SEQ ID NO.10 [4] The biological hair shape modifying composition according to any one of [1] to [3], wherein the alkaline agent component includes a plant-derived alkaline agent and / or a chemically synthesized alkaline agent. [5] The biological hair shape modifying composition according to any one of [1] to [4], wherein the concentration of the alkaline agent component in the biological hair shape modifying composition is 1 to 80% (v / v). [6] The biological hair shape modifying composition according to any one of [1] to [5], wherein the Bacillus licheniformis includes Bacillus licheniformis BCRC 14353, Bacillus licheniformis GI_5E-1 having deposit number BCRC 911027, or a combination thereof. [7] The biological hair shape modifying composition according to any of [1] to [6], wherein the keratin and / or keratin polymer-containing medium comprises a keratin and / or keratin polymer-containing substrate, and the keratin and / or keratin polymer-containing substrate comprises animal feathers, hair, hooves, claws and / or horns. [8] The biological hair shape modifying composition according to any one of [1] to [7], wherein the volume ratio of the polymer component to the small molecule component in the hair shape modifying composition is 5-10:1.5-3. [9] A biological hair shape modification kit, comprising: a polymer component comprising a protease belonging to the alkaline protease class; a small molecule component comprising a peptide having reducing activity; An alkaline agent component; at least one packaging container for containing the polymer component, the small molecule component and the alkaline agent component; Including, both the macromolecular component and the small molecule component are obtained from the fermentation product of Bacillus licheniformis with a medium containing keratin and / or a keratin polymer; The molecular weight of the component in the polymer component is 3 kDa or more and is 3 to 1000 kDa, and the molecular weight of the component in the small molecule component is less than 3 kDa and is 0.01 to 2.99 kDa; The biological hair shape modification kit is used in the form of a mixture of the polymer component, the small molecule component and the alkaline agent component.

[10] The biological hair shape modification kit according to [9], wherein the protease belonging to the alkaline protease class includes alkaline protease, keratinase, or a combination thereof.

[11] The biological hair shape modification kit described in [9] or

[10] , wherein the peptide having reducing activity includes at least one of the following peptides: (a) a peptide comprising the amino acid sequence SEQ ID NO.1: (b) a peptide comprising the amino acid sequence SEQ ID NO.2; (c) a peptide comprising the amino acid sequence SEQ ID NO.3; (d) a peptide comprising the amino acid sequence SEQ ID NO.4; (e) a peptide comprising the amino acid sequence SEQ ID NO.5; (f) a peptide comprising the amino acid sequence SEQ ID NO.6; (g) a peptide comprising the amino acid sequence SEQ ID NO.7; (h) a peptide comprising the amino acid sequence SEQ ID NO.8; (i) a peptide comprising the amino acid sequence SEQ ID NO.9; and (j) a peptide comprising the amino acid sequence SEQ ID NO.10

[12] The biological hair shape modification kit according to any one of [9] to

[11] , wherein the alkaline agent component includes a plant-derived alkaline agent and / or a chemically synthesized alkaline agent.

[13] The biological hair shape modification kit according to

[12] , wherein the plant-derived alkaline agent includes at least one of the following plant-derived alkaline agents: an aqueous extract of coffee grounds, an aqueous extract of tea stems, and an aqueous extract of mulberry leaves.

[14] The biological hair shape modification kit according to any one of [9] to

[13] , wherein the Bacillus licheniformis includes Bacillus licheniformis BCRC 14353, Bacillus licheniformis GI_5E-1 having deposit number BCRC 911027, or a combination thereof.

[15] A biological hair shape modification kit described in any of [9] to

[14] , wherein the keratin and / or keratin polymer-containing medium contains a keratin and / or keratin polymer-containing substrate, and the keratin and / or keratin polymer-containing substrate includes animal feathers, hair, hooves, claws and / or horns.

[16] The biological hair shape modification kit according to

[15] , wherein the content of the keratin and / or keratin polymer-containing substrate in the keratin and / or keratin polymer-containing medium is 0.01 to 20 g / L.

[17] In the biological hair shape modification kit, the polymer component is contained in a first reagent, the small molecule component is contained in a second reagent, and the alkaline agent component is contained in a third reagent; the at least one packaging container includes a first container, a second container, and a third container, the first reagent is packaged in the first container, the second reagent is packaged in the second container, and the third reagent is packaged in the third container; The biological hair shape modification kit is used by mixing the first reagent, the second reagent, and the third reagent. A biological hair shape modification kit according to any one of [9] to

[16] .

[18] The biological hair shape modification kit is used by mixing the first reagent, the second reagent, the third reagent and additional water in a volume ratio of 5 to 10: 1.5 to 3: 3 to 6: 1 to 10.5.

[17] A biological hair shape modification kit as described in the above.

[19] The biological hair shape modification kit is used by pouring out all of the first reagent, the second reagent, and the third reagent from the first container, the second container, and the third container, respectively, and mixing them with additional water.

[17] A biological hair shape modification kit as described in the above.

[20] In the biological hair shape modification kit, the polymer component and the small molecule component are contained in a first reagent, and the alkaline agent component is contained in a second reagent; the at least one packaging container includes a first container and a second container, the first reagent is packaged in the first container, and the second reagent is packaged in the second container; The biological hair shape modification kit is used by mixing the first reagent and the second reagent. A biological hair shape modification kit according to any one of [9] to

[16] .

[21] The biological hair shape modification kit described in

[20] , wherein the biological hair shape modification kit is used by mixing the first reagent, the second reagent, and additional water in a volume ratio of 6.5 to 13:3 to 6:1 to 10.5.

[22] The biological hair shape modification kit described in

[20] , wherein the biological hair shape modification kit is used by pouring out all of the first reagent and the second reagent from the first container and the second container, respectively, and mixing them with additional water.

[23] In the biological hair shape modification kit, the polymer component, the small molecule component and the alkaline agent component are contained in a first reagent; The at least one packaging container includes a first container, and the first reagent is packaged in the first container; A biological hair shape modification kit according to any one of [9] to

[16] .

[24] A method for changing hair shape, comprising: (a) a hair penetration step of applying a mixture produced using the biological hair shape modification composition according to any one of [1] to [8] or the biological hair shape modification kit according to any one of [9] to

[23] to hair and allowing the mixture to penetrate into the hair; (b) a hair shape modification step of subjecting the hair to which the biological hair shape modification composition or the mixture has been applied and permeated, to a heating process to hydrolyze the protein backbone of the hair and cleave disulfide bonds in the protein of the hair, thereby modifying the shape of the hair; A method for altering hair shape comprising:

[25] The method for changing a hair shape according to

[24] , wherein the temperature of the heating step is 40 to 100°C. The following embodiments will be described in detail with reference to the accompanying drawings. [Brief description of the drawings]

[0010] The invention can be more fully understood from the following detailed description and examples, taken in conjunction with the accompanying drawings, in which: [Figure 1] The enzyme activities of the fermentation broth produced by Bacillus licheniformis GI_5E-1 at different temperatures are shown. [Diagram 2]The enzyme activity of the fermentation broth produced by Bacillus licheniformis GI_5E-1 at different pH values ​​is shown. [Diagram 3] Figure 2 shows the disulfide bond cleavage efficiency of the fermentation broth produced by Bacillus licheniformis GI_5E-1 at different temperatures. [Figure 4] The disulfide bond cleavage efficiency and reducing activity of the fermentation concentrate produced by Bacillus licheniformis BCRC 14353 or the fermentation concentrate produced by Bacillus licheniformis GI_5E-1 are shown. [Diagram 5] The results of the skin irritation analysis of the fermentation concentrate produced by Bacillus licheniformis GI_5E-1 are shown. [Figure 6] The results of the skin sensitization analysis of the fermentation concentrate produced by Bacillus licheniformis GI_5E-1 are shown. [Figure 7] The results of the eye irritation analysis of the fermentation concentrate produced by Bacillus licheniformis GI_5E-1 are shown. [Figure 8] The results of stability analysis of the fermentation concentrate produced by Bacillus licheniformis GI_5E-1 are shown. [Figure 9] The results of electrophoretic analysis of the polymer fraction of the fermentation broth produced by Bacillus licheniformis GI_5E-1 are shown. [Figure 10] The reducing activity of 10 peptides contained in the small molecule isolated solution of the fermentation concentrate produced by Bacillus licheniformis GI_5E-1 is shown. [Figure 11]Figure 2 shows the disulfide bond cleavage efficiency of the mixture of water extract of tea stem, water extract of mulberry leaves or water extract of coffee grounds with fermentation liquid produced by Bacillus licheniformis GI_5E-1. [Figure 12] Figure 2 shows the disulfide bond cleavage efficiency of mixtures of water extract of coffee grounds with different amounts of added water and fermentation liquid produced by Bacillus licheniformis GI_5E-1. [Figure 13] 1 shows an analysis of the disulfide bond cleavage efficiency of hair shape altering compositions of different formulations. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0011] In the following detailed description, for purposes of explanation, numerous specific details are set forth in order to provide a thorough understanding of the disclosed embodiments. However, it will be apparent that one or more embodiments may be practiced without these specific details. In addition, for purposes of simplicity, well-known structures and devices are generally shown.

[0012] The present disclosure can provide a biological hair shape modification composition that is hypoallergenic or non-irritating, hypoallergenic or non-allergenic, and has excellent hair shape modification ability. The hair described herein may include any animal hair, and may be hair growing on the skin of an animal and / or hair separated from an animal, without any particular limitation. In one embodiment, the hair described herein may be human hair, and in particular may be hair growing on the skin of a human.

[0013] The biological hair shape modifying compositions of the present disclosure described above may include, but are not limited to, a polymer component, a small molecule component, and an alkaline agent component.

[0014] Both the macromolecular and small molecule components in the biological hair shape modifying compositions of the present disclosure described above may be obtained from the fermentation product of Bacillus licheniformis and a medium containing keratin and / or keratin polymers.

[0015] There are no particular limitations on the polymer composition components in the biological hair shape modifying composition of the present disclosure as described above, as long as they have the effect of hydrolyzing the protein backbone (carbon-carbon chain) of hair, and the effect of the polymer composition components in the biological hair shape modifying composition of the present disclosure is not limited to only hydrolyzing the protein backbone (carbon-carbon chain) of hair.

[0016] In addition, the polymer component in the biological hair shape modifying composition of the present disclosure described above has excellent heat resistance, acid resistance, and alkali resistance. The polymer component in the biological hair shape modifying composition of the present disclosure described above can maintain its effect over a wide temperature range, for example, about 10 to 100° C. Furthermore, the polymer component in the biological hair shape modifying composition of the present disclosure described above can maintain its effect over a wide pH range, for example, about pH 3 to 12.

[0017] In one embodiment, the molecular weight of the component in the polymer component is about 3 kDa or more, for example, about 3.5 kDa or more, about 4 kDa or more, about 5 kDa or more, about 10 kDa or more, about 50 kDa or more, about 100 kDa or more, about 1000 kDa or more, about 3 to 1000 kDa, about 5 to 500 kDa, about 10 to 100 kDa, about 5 ... Da, about 10 kDa, about 15 kDa, about 20 kDa, about 30 kDa, about 40 kDa, about 50 kDa, about 60 kDa, about 63 kDa, about 65 kDa, about 70 kDa, about 80 kDa, about 90 kDa, about 100 kDa, about 200 kDa, about 300 kDa, about 400 kDa, about 500 kDa, but is not limited thereto. In one embodiment, the molecular weight of the components in the above-mentioned polymer component may be about 3 to 1000 kDa.

[0018] The aforementioned polymeric component may include, but is not limited to, a protease belonging to the alkaline protease family. The aforementioned proteases belonging to the alkaline protease family may have the effect of hydrolyzing the protein backbone (carbon-carbon chain) of hair, but are not limited thereto. In one embodiment, the aforementioned polymeric component may include, but is not limited to, an alkaline protease, a keratinase, a serine protease, a tyrosine protease, and the like, or any combination thereof. In one embodiment, the aforementioned polymeric component may include an alkaline protease. In another embodiment, the aforementioned polymeric component may include a keratinase. In yet another embodiment, the aforementioned polymeric component may include a combination of an alkaline protease and a keratinase.

[0019] The molecular weight of the protease belonging to the alkaline proteases is about 3 kDa or more, for example, about 3.5 kDa or more, about 4 kDa or more, about 5 kDa or more, about 10 kDa or more, about 50 kDa or more, about 100 kDa or more, about 1000 kDa, about 3 to 1000 kDa, about 5 to 500 kDa, about 10 to 100 kDa, about 5 kDa, The molecular weight of the protease may be, but is not limited to, about 10 kDa, about 15 kDa, about 20 kDa, about 30 kDa, about 40 kDa, about 50 kDa, about 60 kDa, about 63 kDa, about 65 kDa, about 70 kDa, about 80 kDa, about 90 kDa, about 100 kDa, about 200 kDa, about 300 kDa, about 400 kDa, about 500 kDa, etc. In one embodiment, the molecular weight of the protease belonging to the alkaline proteases may be about 3 to 1000 kDa. In another embodiment, the molecular weight of the protease belonging to the alkaline proteases may be about 63 kDa. In a particular embodiment, the protease belonging to the alkaline proteases is an alkaline protease, keratinase, or a combination thereof, having a molecular weight of about 63 kDa.

[0020] Furthermore, the content of alkaline protease in the biological hair shape modifying composition of the present disclosure described above may be about 150 to 3000 U / mL, for example, about 200 U / mL, about 215 U / mL, about 230 U / mL, about 250 U / mL, about 300 U / mL, about 315 U / mL, about 320 U / mL, about 330 U / mL, about 333 U / mL, about 350 U / mL, about 400 U / mL, about 500 U / mL, about 1000 U / mL, about 1500 U / mL, about 2000 U / mL, about 2500 U / mL, or about 3000 U / mL, but is not limited thereto.

[0021] The small molecule component in the biological hair shape modifying composition of the present disclosure described above is also not particularly limited as long as it is capable of cleaving disulfide bonds in hair proteins, and the effect of the small molecule component in the biological hair shape modifying composition of the present disclosure described above is not limited to cleaving disulfide bonds in hair proteins.

[0022] The molecular weight of the components in the small molecule component described above is less than about 3 kDa, for example, less than about 2.99 kDa, less than about 2.5 kDa, less than about 2 kDa, less than about 1 kDa, less than about 0.5 kDa, about 0.01 to 2.99 kDa, about 0.1 to 2.8 kDa, about 0.5 to 2.7 kDa, about 1 to 2.6 kDa, about 1.2 to 2.5 kDa, about 0.5 kDa, about 0.55 kDa, about 1.1 kDa, about 1.2 kDa, or about 1.5 kDa. The molecular weight of the small molecule component may be, but is not limited to, about 1.3 kDa, about 1.4 kDa, about 1.5 kDa, about 1.6 kDa, about 1.7 kDa, about 1.8 kDa, about 1.9 kDa, about 2.0 kDa, about 2.1 kDa, about 2.2 kDa, about 2.3 kDa, about 2.4 kDa, about 2.5 kDa, about 2.6 kDa, about 2.7 kDa, about 2.8 kDa, about 2.9 kDa, about 2.95 kDa, or about 2.99 kDa. In one embodiment, the molecular weight of the component of the small molecule component described above may be about 0.01 to 2.99 kDa.

[0023] The small molecule component may include, but is not limited to, a peptide having reducing activity. In addition to reducing activity, the peptide having reducing activity can also cleave disulfide bonds in hair proteins, but the effect is not limited thereto.

[0024] The length of the peptide having the above-mentioned reducing activity may be about 5 to 30 amino acids, for example, about 6 to 28 amino acids, about 11 to 25 amino acids, about 10 amino acids, about 11 amino acids, about 12 amino acids, about 13 amino acids, about 14 amino acids, about 15 amino acids, about 20 amino acids, about 23 amino acids, about 25 amino acids, or about 30 amino acids, but is not limited thereto.

[0025] In addition, the peptides having the above-mentioned reducing activity may include, but are not limited to, at least one of the following peptides: (a) a peptide comprising the amino acid sequence SEQ ID NO. (b) a peptide comprising the amino acid sequence SEQ ID NO.2 (c) a peptide comprising the amino acid sequence SEQ ID NO.3; (d) a peptide comprising the amino acid sequence SEQ ID NO.4; (e) a peptide comprising the amino acid sequence SEQ ID NO.5; (f) a peptide comprising the amino acid sequence SEQ ID NO.6; (g) a peptide comprising the amino acid sequence SEQ ID NO.7; (h) a peptide comprising the amino acid sequence SEQ ID NO.8; (i) a peptide comprising the amino acid sequence SEQ ID NO.9; and (j) A peptide comprising the amino acid sequence SEQ ID NO.10.

[0026] In one embodiment, the small molecule component may include the peptide comprising the amino acid sequence SEQ ID NO.2. In another embodiment, the small molecule component may include the peptide comprising the amino acid sequence SEQ ID NO.3. In yet another embodiment, the small molecule component may include the peptide comprising the amino acid sequence SEQ ID NO.6. In yet another embodiment, the small molecule component may include the peptide comprising the amino acid sequence SEQ ID NO.10. In a particular embodiment, the small molecule component may include the peptide comprising the amino acid sequence SEQ ID NO.2, the peptide comprising the amino acid sequence SEQ ID NO.3, the peptide comprising the amino acid sequence SEQ ID NO.6, and the peptide comprising the amino acid sequence SEQ ID NO.10. In another specific embodiment, the small molecule component may include the peptide comprising the amino acid sequence of SEQ ID NO.1, the peptide comprising the amino acid sequence of SEQ ID NO.2, the peptide comprising the amino acid sequence of SEQ ID NO.3, the peptide comprising the amino acid sequence of SEQ ID NO.4, the peptide comprising the amino acid sequence of SEQ ID NO.5, the peptide comprising the amino acid sequence of SEQ ID NO.6, the peptide comprising the amino acid sequence of SEQ ID NO.7, the peptide comprising the amino acid sequence of SEQ ID NO.8, the peptide comprising the amino acid sequence of SEQ ID NO.9, and the peptide comprising the amino acid sequence of SEQ ID NO.10.

[0027] In addition, the fermentation products of the aforementioned Bacillus licheniformis with keratin and / or keratin polymer-containing media from which the macromolecular and small molecule components of the biological hair shape modifying compositions of the present disclosure can be obtained are described in more detail below.

[0028] There is no particular limitation on the Bacillus licheniformis involved in the fermentation product of the aforementioned Bacillus licheniformis with a medium containing keratin and / or a keratin polymer, so long as the macromolecular and small molecule components of the biological hair shape modifying composition of the present disclosure can be obtained therefrom. The aforementioned Bacillus licheniformis may include, but is not limited to, Bacillus licheniformis BCRC 14353, Bacillus licheniformis GI_5E-1 deposited at the Bioresource Collection and Research Center (BCRC) of Food industry research and development institute (FIRDI) of the Republic of China (Taiwan) on November 26, 2020 under the accession number BCRC 911027, and the like, or any combination thereof. In one embodiment, the Bacillus licheniformis involved in the fermentation product of the Bacillus licheniformis and the medium containing keratin and / or keratin polymer may be Bacillus licheniformis BCRC 14353. In another embodiment, the Bacillus licheniformis involved in the fermentation product of the Bacillus licheniformis and a keratin and / or keratin polymer-containing medium described above may be Bacillus licheniformis GI_5E-1, deposited at the Food Industry Research Institute (FIRDI) Bioresources Conservation and Research Center (BCRC) of the Republic of China (Taiwan) on November 26, 2020 under the deposit number BCRC 911027.

[0029] Furthermore, the above-mentioned keratin and / or keratin polymer-containing medium may contain, but is not limited to, a keratin and / or keratin polymer-containing substrate. The content of the keratin and / or keratin polymer-containing substrate in the keratin and / or keratin polymer-containing medium may be, but is not limited to, about 0.01 to 20 g / L, for example, about 0.02 to 15 g / L, about 0.05 to 10 g / L, about 1 to 5 g / L, about 0.01 g / L, about 0.05 g / L, about 0.1 g / L, about 0.5 g / L, about 1 g / L, about 2 g / L, about 4 g / L, about 5 g / L, about 6 g / L, about 8 g / L, or about 10 g / L. In one embodiment, the content of the keratin and / or keratin polymer-containing substrate in the keratin and / or keratin polymer-containing medium may be about 10 g / L.

[0030] In addition, the aforementioned keratin and / or keratin polymer-containing substrate may include, but is not limited to, animal feathers, hair, hooves, claws, horns, etc., or any combination thereof. The aforementioned animal feathers, hair, hooves, claws, horns, etc. may be discarded animal feathers, hair, hooves, claws, horns, etc. In the case where the aforementioned keratin and / or keratin polymer-containing substrate may include discarded animal feathers, hair, hooves, claws, horns, etc., or any combination thereof, since the macromolecular components and small molecule components in the biological hair shape modifying composition of the present disclosure described above are obtained from waste, the method of obtaining the macromolecular components and small molecule components in the biological hair shape modifying composition of the present disclosure can be regarded as a method of recycling and reusing waste, which has an effect on environmental protection. In one embodiment, the aforementioned keratin and / or keratin polymer-containing substrate may include feathers. In a particular embodiment, the aforementioned keratin and / or keratin polymer-containing substrate may include discarded feathers. In this particular embodiment, where discarded feathers may be included in the keratin and / or keratin polymer-containing substrate, the content of discarded feathers in the keratin and / or keratin polymer-containing medium may be, but is not limited to, about 0.01-20 g / L, for example, about 0.02-15 g / L, about 0.05-10 g / L, about 1-5 g / L, about 0.01 g / L, about 0.05 g / L, about 0.1 g / L, about 0.5 g / L, about 1 g / L, about 2 g / L, about 4 g / L, about 5 g / L, about 6 g / L, about 8 g / L, or about 10 g / L.

[0031] Furthermore, the fermentation product of the above-mentioned Bacillus licheniformis and the keratin and / or keratin polymer-containing medium can be obtained by culturing the above-mentioned Bacillus licheniformis in the above-mentioned keratin and / or keratin polymer-containing medium. The temperature at which the above-mentioned Bacillus licheniformis is cultured may be about 25 to 70°C, for example, about 30 to 45°C, about 45 to 55°C, about 55 to 65°C, about 30°C, about 35°C, about 37°C, about 40°C, about 45°C, about 50°C, about 55°C, about 60°C, or about 65°C, but is not limited thereto. The time for culturing the above-mentioned Bacillus licheniformis may be about 24 to 96 hours, for example, about 48 to 72 hours, about 24 hours, about 48 hours, about 72 hours, or about 96 hours, but is not limited thereto.

[0032] The polymeric and small molecule components in the biological hair shape modifying composition of the present disclosure described above can be obtained by, but is not limited to, obtaining a fermentation product of the aforementioned Bacillus licheniformis with a medium containing keratin and / or a keratin polymer, and then filtering the fermentation product through a membrane capable of separating components having a molecular weight of 3 kDa or more from components having a molecular weight of less than 3 kDa.

[0033] In addition, the small molecule component can be further mixed with the new keratin and / or keratin polymer-containing substrate to form a mixture and then cultured, thereby enhancing the reducing activity of the small molecule component and / or the effect of breaking the disulfide bonds of hair proteins. The description of the new keratin and / or keratin polymer-containing substrate described here is the same as the description of the keratin and / or keratin polymer-containing substrate described above, so it will not be repeated here. The content of the new keratin and / or keratin polymer-containing substrate in the above-mentioned mixture is about 0.05 to 2% (w / v), for example, about 0.1 to 1% (w / v), about 0.2 to 0.8% (w / v), about 0.1% (w / v), about 0.2% (w / v), about 0.5% (w / v), about 0.8% (w / v), about 1% (w / v), but is not limited thereto.

[0034] The alkaline agent component in the biological hair shape modifying compositions of the present disclosure may include, but is not limited to, plant-derived alkaline agents, chemically synthetically derived alkaline agents, or combinations thereof.

[0035] The concentration of the alkaline agent component in the biological hair shape modifying composition of the present disclosure may be, but is not limited to, about 1 to 80% (v / v), for example, about 5 to 60% (v / v), about 10 to 50% (v / v), about 5% (v / v), about 10% (v / v), about 15% (v / v), about 25% (v / v), about 30% (v / v), or about 50% (v / v).

[0036] The aforementioned plant-derived alkaline agent is not particularly limited, and may be any alkaline agent derived from a plant source. The aforementioned plant-derived alkaline agent may have the effect of increasing the efficiency of disulfide bond cleavage of small molecule components, but is not limited thereto. The aforementioned plant-derived alkaline agent may include, but is not limited to, at least one of the following plant-derived alkaline extracts: coffee grounds extract, tea stem extract, mulberry leaf extract, etc., or any combination thereof. The aforementioned plant-derived extract may be, but is not limited to, a plant-derived aqueous extract. In one embodiment, the aforementioned plant-derived alkaline agent may include, but is not limited to, at least one of the following plant-derived extracts: coffee grounds aqueous extract, tea stem aqueous extract, mulberry leaf aqueous extract, etc. The coffee grounds aqueous extract may include, but is not limited to, nicotinic acid, etc. The tea stem aqueous extract may include, but is not limited to, theophylline, and the mulberry leaf aqueous extract may include, but is not limited to, a plant alkaloid. In a particular embodiment, the alkaline agent component in the biological hair shape modifying composition of the present disclosure can be a water extract of coffee grounds.Coffee grounds, tea stems, etc. are usually considered waste, and the alkaline agent of the biological hair shape modifying composition of the present disclosure can be obtained from waste, so that the waste can be recycled and reused.Therefore, the biological hair shape modifying composition of the present disclosure can have the effect of environmental protection.

[0037] In addition, the aforementioned chemically synthesized alkaline agents may include, but are not limited to, at least one of the following ingredients: nicotinic acid, theophylline, plant alkaloids, ammonia, etc.

[0038] Furthermore, in the biological hair shape modifying composition of the present disclosure, the volume ratio of the aforementioned polymer component to the aforementioned small molecule component may be, but is not limited to, about 5-10:1.5-3. In one embodiment, in the biological hair shape modifying composition of the present disclosure, the volume ratio of the aforementioned polymer component to the aforementioned small molecule component may be about 5:1.5. In another embodiment, in the biological hair shape modifying composition of the present disclosure, the volume ratio of the aforementioned polymer component to the aforementioned small molecule component may be about 10:3.

[0039] In one embodiment, the biological hair shape modifying composition of the present disclosure may further comprise additional water, which may include, but is not limited to, tap water, RO water, purified water, and the like.

[0040] The present disclosure can also provide a biological hair shape modification kit that is used to modify hair shape, is hypoallergenic or non-irritating, hypoallergenic or non-allergenic, and has excellent hair shape modification ability. The hair described herein may include any animal hair, and may be hair growing on the skin of an animal and / or hair separated from an animal, without any particular limitation. In one embodiment, the hair described herein may be human hair, and in particular may be hair growing on the skin of a human.

[0041] The biological hair shape modification kit of the present disclosure described above may include, but is not limited to, a polymer component, a small molecule component, an alkali component, and at least one packaging container.In addition, the biological hair shape modification kit of the present disclosure may be used in the form of a mixture of the above-mentioned polymer component, the above-mentioned small molecule component, and the above-mentioned alkali agent component.

[0042] The polymer component, small molecule component and alkaline agent component in the biological hair shape modifying kit of the present disclosure described above may be the same as the polymer component, small molecule component and alkaline agent component in the biological hair shape modifying composition of the present disclosure described above, so the description thereof will not be repeated here.

[0043] At least one packaging container in the biological hair shape modification kit of the present disclosure is used to contain the polymer composition component, the small molecule composition component, and the alkaline composition component. There is no particular limitation on the material of the at least one packaging container described above, and it is sufficient that it is not affected by the polymer composition component, the small molecule composition component, the alkaline agent component, or any combination thereof, and / or does not adversely affect the effects of the polymer composition component, the small molecule composition component, the alkaline agent component, or any combination thereof. The material of the at least one packaging container described above may include, but is not limited to, glass, plastic, ceramic, etc., a composite material made of any of the above materials, or any combination of the above materials.

[0044] In one embodiment, in the biological hair shape modification kit of the present disclosure described above, the above-mentioned macromolecular component may be contained in the first reagent, the above-mentioned small molecule component may be contained in the second reagent, the above-mentioned alkaline agent component may be contained in the third reagent, and the above-mentioned at least one packaging container may include a first container, a second container, and a third container. The above-mentioned first reagent may be packaged in the above-mentioned first container, the above-mentioned second reagent may be packaged in the above-mentioned second container, and the above-mentioned third reagent may be packaged in the above-mentioned third container. Furthermore, in this embodiment, the biological hair shape modification kit of the present disclosure may be used in the form of a mixture of the above-mentioned first reagent, the above-mentioned second reagent, and the above-mentioned third reagent. In one use scenario of this embodiment, the above-mentioned biological hair shape modification kit of the present disclosure can be used by mixing the above-mentioned first reagent, the above-mentioned second reagent, the above-mentioned third reagent, and additional water (e.g., tap water, RO water, pure water, but not limited thereto) in a specific volume ratio. The specific volume ratios mentioned above may be, but are not limited to, about 5 to 10:1.5 to 3:3 to 6:1 to 10.5, for example, about 5:1.5:3:10.5, about 5:1.5:6:7.5, about 10:3:6:1. In another use scenario of this embodiment, the biological hair shape modification kit of the present disclosure described above can be used by pouring out all of the first reagent, the second reagent, and the third reagent from the first container, the second container, and the third container, respectively, and mixing them with additional water.In yet another usage scenario of this embodiment, the biological hair shape modification kit of the present disclosure described above can be used by pouring out all of the aforementioned first reagent and the aforementioned second reagent from the aforementioned first container and the aforementioned second container into the aforementioned third container, respectively, and adding water to the aforementioned third container so that the aforementioned first reagent, the aforementioned second reagent, the aforementioned third reagent and water are mixed in the aforementioned third container; or pouring out all of the aforementioned first reagent and the aforementioned third reagent from the aforementioned first container and the aforementioned third container into the aforementioned third container, respectively, and adding water to the aforementioned third container so that the aforementioned first reagent, the aforementioned second reagent, the aforementioned third reagent and water are mixed in the aforementioned third container. and pouring the first reagent, the second reagent, the third reagent, and water into the second container so that the first reagent, the second reagent, the third reagent, and water are mixed in the second container, or pouring all of the second reagent and the third reagent from the second container and the third container into the first container, respectively, and adding water into the first container so that the first reagent, the second reagent, the third reagent, and water are mixed in the first container. In addition, in this embodiment, the biological hair shape modification kit of the present disclosure may further include a mixing container for containing and mixing the first reagent, the second reagent, and the third reagent, and water, which are poured out from the first container, the second container, and the third container, respectively. The material of the mixing container is not particularly limited, and may be any material that is not affected by the mixture of the first reagent, the second reagent, the third reagent, and water during use of the kit, and / or does not adversely affect the effects of the mixture of the first reagent, the second reagent, the third reagent, and water. The material of the mixing container may include glass, plastic, ceramic, paper, etc., a composite material made of any of the above materials, or any combination of the above materials.

[0045] In another embodiment, in the biological hair shape modification kit of the present disclosure described above, the above-mentioned polymer component and the above-mentioned small molecule component may be contained in the first reagent, the above-mentioned alkaline agent component may be contained in the second reagent, the above-mentioned at least one packaging container may include a first container and a second container, the above-mentioned first reagent may be packaged in the above-mentioned first container, and the above-mentioned second reagent may be packaged in the above-mentioned second container. Furthermore, in this embodiment, the biological hair shape modification kit of the present disclosure may be used in the form of a mixture of the above-mentioned first reagent and the above-mentioned second reagent. In one use scenario of this embodiment, the above-mentioned biological hair shape modification kit of the present disclosure may be used by mixing the above-mentioned first reagent, the above-mentioned second reagent, and additional water (e.g., tap water, RO water, pure water, but not limited thereto) in a specific volume ratio. The aforementioned specific volume ratio may be, but is not limited to, about 6.5 to 13:3 to 6:1 to 10.5, for example, about 6.5:3:10.5, about 6.5:6:7, 5, about 13:6:1. In another usage scenario of this embodiment, the biological hair shape modification kit of the present disclosure described above can be used by pouring out all of the first and second reagents from the first and second containers, respectively, and mixing them with additional water. In another use scenario of this embodiment, the biological hair shape modification kit of the present disclosure described above can be used by pouring out all of the first reagent from the first container into the second container, and adding water to the second container so that the first reagent, the second reagent and the water are mixed in the second container, or pouring out all of the second reagent from the second container into the first container, and adding water to the first container so that the first reagent, the second reagent and the water are mixed in the first container.In addition, in this embodiment, the biological hair shape modification kit of the present disclosure may further include a mixing container for containing and mixing the first reagent and the second reagent, and water, which are poured out from the first container and the second container, respectively.The material of the mixing container is not particularly limited, and may be any material that is not affected by the mixture of the first reagent, the second reagent, and water during use of the kit, and / or does not adversely affect the effects of the mixture of the first reagent, the second reagent, and water. The material of the mixing container may include glass, plastic, ceramic, paper, etc., a composite material made of any of the above materials, or any combination of the above materials.

[0046] In another embodiment, in the biological hair shape modification kit of the present disclosure described above, the above-mentioned polymer component may be contained in the first reagent, the above-mentioned small molecule component and the above-mentioned alkaline agent component may be contained in the second reagent, the above-mentioned at least one packaging container may include a first container and a second container, the above-mentioned first reagent may be packaged in the above-mentioned first container, and the above-mentioned second reagent may be packaged in the above-mentioned second container. Furthermore, in this embodiment, the biological hair shape modification kit of the present disclosure may be used in the form of a mixture of the above-mentioned first reagent and the above-mentioned second reagent. In one use scenario of this embodiment, the above-mentioned biological hair shape modification kit of the present disclosure can be used by mixing the above-mentioned first reagent, the above-mentioned second reagent, and additional water (e.g., tap water, RO water, pure water, but not limited to these) in a specific volume ratio. The specific volume ratio mentioned above may be, but is not limited to, about 1 to 10:0.9 to 9:1 to 2.1, for example, about 1:0.9:2.1, about 1:1.5:1.5, about 10:9:1. In another usage scenario of this embodiment, the biological hair shape modification kit of the present disclosure described above can be used by pouring out all of the first and second reagents from the first and second containers, respectively, and mixing them with additional water. In another use scenario of this embodiment, the biological hair shape modification kit of the present disclosure described above can be used by pouring out all of the first reagent from the first container into the second container, and adding water to the second container so that the first reagent, the second reagent, and the water are mixed in the second container, or pouring out all of the second reagent from the second container into the first container, and adding water to the first container so that the first reagent, the second reagent, and the water are mixed in the first container.In addition, in this embodiment, the biological hair shape modification kit of the present disclosure may further include a mixing container for containing and mixing the first reagent and the second reagent and water poured out from the first container and the second container, respectively.The material of the mixing container is not particularly limited, and may be any material that is not affected by the mixture of the first reagent, the second reagent, and water during use of the kit, and / or does not adversely affect the effects of the mixture of the first reagent, the second reagent, and water. The material of the mixing container may include glass, plastic, ceramic, paper, etc., a composite material made of any of the above materials, or any combination of the above materials.

[0047] In another embodiment, in the biological hair shape modification kit of the present disclosure described above, the above-mentioned polymer component and the above-mentioned alkaline agent component may be contained in the first reagent, the above-mentioned small molecule component may be contained in the second reagent, the above-mentioned at least one packaging container may include a first container and a second container, the above-mentioned first reagent may be packaged in the above-mentioned first container, and the above-mentioned second reagent may be packaged in the above-mentioned second container. Furthermore, in this embodiment, the biological hair shape modification kit of the present disclosure described above of the first reagent and the above-mentioned second reagent may be used in the form of a premix. In one use scenario of this embodiment, the biological hair shape modification kit of the present disclosure described above can be used by mixing the above-mentioned first reagent, the above-mentioned second reagent, and additional water (e.g., tap water, RO water, pure water, but not limited thereto) in a specific volume ratio. The aforementioned specific volume ratio may be, but is not limited to, about 8 to 16:1.5 to 3:1 to 10.5, for example, about 8:1.5:10.5, about 11:1.5:7.5, about 16:3:1. In another usage scenario of this embodiment, the above-mentioned biological hair shape modification kit of the present disclosure can be used by pouring out all of the first and second reagents from the first and second containers, respectively, and mixing them with additional water. In another use scenario of this embodiment, the biological hair shape modification kit of the present disclosure described above can be used by pouring out all of the first reagent from the first container into the second container, and adding water to the second container so that the first reagent, the second reagent, and the water are mixed in the second container, or pouring out all of the second reagent from the second container into the first container, and adding water to the first container so that the first reagent, the second reagent, and the water are mixed in the first container.In addition, in this embodiment, the biological hair shape modification kit of the present disclosure may further include a mixing container for containing and mixing the first reagent and the second reagent and the water poured out from the first container and the second container, respectively.The material of the mixing container is not particularly limited, and may be any material that is not affected by the mixture of the first reagent, the second reagent, and water during use of the kit, and / or does not adversely affect the effects of the mixture of the first reagent, the second reagent, and water. The material of the mixing container may include glass, plastic, ceramic, paper, etc., a composite material made of any of the above materials, or any combination of the above materials.

[0048] Furthermore, in one embodiment, in the biological hair shape modification kit of the present disclosure described above, the above-mentioned polymer component, the above-mentioned small molecule component, and the above-mentioned alkaline agent component may be contained in the first reagent, and the above-mentioned at least one packaging container may include a first container. The above-mentioned first reagent may be packaged in the above-mentioned first container. In this embodiment, the above-mentioned biological hair shape modification kit of the present disclosure can be used by using the above-mentioned first reagent as it is, or by mixing the above-mentioned first reagent with water, and the volume ratio of the above-mentioned first reagent to water is about 1:0.05 to 1.1, but is not limited thereto.

[0049] In addition, in one embodiment, the above-mentioned biological hair shape modification kit of the present disclosure may further include an oxidizing agent for fixing the shape of the modified hair. Examples of oxidizing agents applicable to the above-mentioned biological hair shape modification kit of the present disclosure may include, but are not limited to, sodium bromate and hydrogen peroxide. Also, in this embodiment, the oxidizing agent may be packaged in a packaging container different from that for packaging the above-mentioned polymer component, the above-mentioned small molecule component, the above-mentioned alkaline agent component, or any combination thereof.

[0050] In addition, in one embodiment, the biological hair shape modification kit of the present disclosure may further include additional water in addition to the above-mentioned polymer component, the above-mentioned small molecule component, and the above-mentioned alkaline agent component. The above-mentioned additional water may include, but is not limited to, tap water, RO water, pure water, etc.

[0051] Further, based on the foregoing, the present disclosure can also provide a method for changing hair shape that can be performed by any of the above-mentioned biological hair shape modifying compositions of the present disclosure, or by a mixture produced by using any of the above-mentioned biological hair shape modifying kits of the present disclosure. The hair used in the method for changing hair shape of the present disclosure can include any animal hair, and can be hair growing on the skin of an animal and / or hair separated from an animal, without any particular limitation. In one embodiment, the hair described herein can be human hair, and in particular hair growing on the skin of a human.

[0052] Compared with conventional methods for changing hair shape, the method of the present disclosure, using any of the above-mentioned biological hair shape modification compositions of the present disclosure, or a mixture produced by using any of the above-mentioned biological hair shape modification kits of the present disclosure, can achieve the effect of effectively modifying shape at low temperatures, and does not cause or is less likely to cause irritation and / or allergic reactions to the skin, eyes, etc. of the animal body, even when the hair is growing on the animal body.

[0053] The method for changing hair shape of the present disclosure described above may include, but is not limited to, a hair infiltrating step and a hair shape modifying step.

[0054] In the aforementioned hair penetration step, any of the above-mentioned biological hair shape modifying compositions of the present disclosure or any of the above-mentioned biological hair shape modifying kits of the present disclosure can be applied to the hair and penetrated therein.

[0055] The time for carrying out the aforementioned hair penetration step can be based on the properties of the hair to be treated (e.g., thickness, hardness, and curliness), the current state of the hair to be treated (e.g., health, dryness), the amount of hair to be treated, and the environmental conditions for treating the hair (e.g., temperature and humidity), and is not particularly limited, as long as the mixture produced by using the above-mentioned biological hair shape modification composition of the present disclosure or the above-mentioned hair shape modification kit of the present disclosure can be permeated into the hair. In one embodiment, the time for carrying out the aforementioned hair penetration step can be about 1 to 60 minutes, for example, about 5 minutes, about 10 minutes, about 15 minutes, about 20 minutes, about 30 minutes, and about 60 minutes, but is not limited thereto.

[0056] Furthermore, the above-mentioned hair infiltration step can be performed at room temperature, but is not limited to this.

[0057] In addition, in the above-mentioned hair shape modification step, a heating step is performed on the hair to which the mixture produced by using the above-mentioned biological hair shape modification composition of the present disclosure or the above-mentioned biological hair shape modification kit of the present disclosure has been applied and permeated, thereby hydrolyzing the backbone of the hair protein and breaking the disulfide bonds in the hair protein, thereby changing the shape of the hair.

[0058] The temperature of the heating step may be, but is not limited to, about 40 to 100° C., for example, about 40° C., about 50° C., about 60° C., about 70° C., about 90° C., and about 100° C. In one embodiment, the temperature of the heating step may be about 60° C.

[0059] The time for performing the hair shape modification step described above can be based on the properties of the hair to be treated (e.g., thickness, hardness, and curliness), the current state of the hair to be treated (e.g., health, dryness), the amount of hair to be treated, and the environmental conditions for treating the hair (e.g., temperature and humidity), and is not particularly limited as long as it can change the shape of the hair. In one embodiment, the time for performing the hair shape modification step described above may be about 1 to 60 minutes, for example, about 5 minutes, about 10 minutes, about 15 minutes, about 20 minutes, about 30 minutes, and about 60 minutes, but is not limited thereto.

[0060] In one embodiment, the method for altering hair shape of the present disclosure, in addition to the hair infiltrating step and hair shape altering step described above, may further include a step of wetting the hair prior to the hair infiltrating step described above.

[0061] In one embodiment, the method for changing the hair shape of the present disclosure, in addition to the hair infiltrating step and the hair shape modifying step described above, may further include a step of washing the hair after the hair shape modifying step described above.

[0062] In one embodiment, the method for changing the hair shape of the present disclosure may be a perm treatment method. In this embodiment, the method for changing the hair shape of the present disclosure may further include a step of winding the hair around hair curlers before the hair permeation step, in addition to the above-mentioned hair permeation step and hair shape modification step.

[0063] In another embodiment, the method for changing the hair shape of the present disclosure can be a method for hair straightening treatment. In this embodiment, the method for changing the hair shape of the present disclosure can, in addition to the above-mentioned hair penetration step and hair shape modification step, simultaneously carry out the above-mentioned heating step and hair straightening step during the above-mentioned hair shape modification step.

[0064] In one embodiment, the method for changing the hair shape of the present disclosure may further include a step of fixing the hair shape after the hair shape modification step, in addition to the above-mentioned hair permeation step and hair shape modification step. The above-mentioned step of fixing the hair shape may include, but is not limited to, applying an oxidizing agent to the hair whose shape has been changed. The above-mentioned oxidizing agent may include sodium bromate, hydrogen peroxide, etc.

[0065] Working Example

[0066] A. Experimental Method

[0067] 1. Cultivation of Bacterial Strains

[0068] The strains were cultured in LB medium (Difco (registered trademark) LB Broth, Miller; BD), and 2% agar (Bacto (registered trademark) agar; BD) was added to the LB medium to prepare LB culture plates.

[0069] The strains (Bacillus licheniformis BCRC 14353 purchased from the Bioresource Conservation and Research Center (BCRC) of the Food Industry Development Institute (FIRDI), Republic of China (Taiwan), or Bacillus licheniformis GI_5E-1, deposited at the Bioresource Conservation and Research Center (BCRC) of the Food Industry Development Institute (FIRDI), Republic of China (Taiwan) on November 26, 2020, with accession number BCRC 911027) were placed in cryopreservation vials and cultured statically for 24 hours in a 50°C constant temperature shaking incubator (LM~420D; YIH DER).

[0070] Then, a single colony was picked from the culture plate and inoculated into a 50 mL centrifuge tube containing 10 mL of LB culture medium. The centrifuge tube was placed at a 45 degree inclination in a 50°C constant temperature incubator and incubated at 150 rpm for 24 hours to obtain a culture bacterial suspension.

[0071] The bacterial suspension was measured for optical density (OD) at 600 nm using a spectrophotometer (SP-880; Metertech). 600 ) was diluted to 0.08-0.1, and 1 mL of the diluted bacterial suspension was taken and inoculated into a 250 mL Erlenmeyer flask containing 150 mL of feather medium (Lin, X., Lee, CG, Casale, ES, & Shih, JC (1992). Purification and characterization of a keratinase from a feather-degrading Bacillus licheniformis strain. Applied and environmental microbiology, 58 (10), 3271-3275), and incubated at 150 rpm for 72 hours in a 50°C constant temperature shaking incubator to obtain an enzyme production induction culture. The composition of the feather medium is shown in Table 1.

[0072] [Table 1]

[0073] The above-mentioned culture medium was centrifuged at 12,000×g and 4° C. for 10 minutes in a refrigerated centrifuge (Centrifuge 5804 R; Eppendorf) to remove large particles of feather powder and to leave the supernatant. The supernatant was then filtered by suction filtration using 0.8 μm and 0.45 μm membrane filters (Membrane filter; Advantec) in sequence, and then further filtered with a 0.22 μm filter cup (Vacuum Filtra; Biofil) to obtain a filtrate. This filtrate was used as the fermentation stock solution.

[0074] The fermentation stock solution was separated and purified using a protein concentration system (KrosFlo KR2i TFF System; Spectrum) (column model S04-E001-05-N) (capable of filtering molecules with a molecular weight of less than 3 kDa from the hollow fiber membrane of the column) at a flow rate of 500 mL / min; back pressure of 12 psi). The portion of the fermentation stock solution that was retained in the hollow fiber membrane of the column was referred to as the macromolecular separation liquid, while the portion that permeated the hollow fiber membrane of the column was referred to as the small molecule separation liquid.

[0075] The small molecule isolated solution was added to 0.5% feather powder and incubated in a 50°C thermostatic incubator at 150 rpm for 48 hours to increase the content of small molecule products and produce a potency-boosted small molecule isolated solution, which was then concentrated by vacuum concentration to remove some of the water from the potency-boosted small molecule isolated solution.

[0076] 2. Enzyme activity analysis

[0077] Enzyme activity analysis was performed using the azocasein assay. The formulation of the 0.5% azocasein solution is shown in Table 2.

[0078] [Table 2]

[0079] 200 μL of the test solution of an appropriate concentration was added to 800 μL of 0.5% azocasein solution, mixed uniformly to prepare a reaction solution, and then reacted at 65°C. After 30 minutes, 200 μL of 10% trichloroacetic acid (TCA) was quickly added to the reaction solution to stop the reaction. The reaction solution was then centrifuged at 12,000 × g and 4°C for 10 minutes. 500 μL of the supernatant was taken and mixed with 500 μL of 0.2 M NaOH solution to prepare a mixed solution. The OD of this mixed solution was 440 The enzyme activity of the test solution was measured as a function of the concentration of azocasein solution versus OD 440 The calculation was based on the calibration curve.

[0080] OD vs. concentration of azocasein solution 440The calibration curve was plotted as follows: The azocasein solution was diluted by the serial half dilution method, and the OD of each concentration was 440 The OD versus concentration of azocasein solution was measured. 440 Plot the scatter plot of the above and then add a trend line to obtain the equation of the concentration of azocasein solution versus OD440, as well as the R of the trend line. 2 The trend line obtained can be used as a calibration curve if R 2 Values ​​greater than 0.995 were considered acceptable. One unit (U) was defined as the degradation of 1 μg of azocasein per minute, and the enzyme concentration was expressed in U / mL.

[0081] 3. Analysis of Disulfide Bond Cleavage Efficiency

[0082] When a compound having a disulfide bond, 5,5'-disulfanediylbis(2-nitrobenzoic acid) (DTNB), reacts with -SH that appears after the cleavage of disulfide bonds in hair, yellow 2-nitro-5-thiobenzoic acid (TNB) is produced. This allows the efficiency of cleaving disulfide bonds in the test sample hair (the content of -SH that appears after the cleavage of disulfide bonds in hair) to be evaluated by using DTNB and a color reaction.

[0083] 0.05 g of human hair was mixed with 5 mL of the test sample to prepare a reaction solution, which was then reacted at 60°C for 30 minutes. The reaction solution was then filtered to remove the supernatant, and 5 mL of DTNB buffer (0.002 M DTNB dissolved in 0.1 M phosphate buffer (pH = 8)) was added and reacted. After reacting for 5 minutes, the reaction solution was filtered to separate the hair, and the filtrate was analyzed at 412 nm to evaluate the amount of TNB produced, thereby evaluating the efficiency of cleaving the disulfide bonds in the test sample hair.

[0084] 4. Redox Activity Analysis

[0085] Red prussic acid can be reduced to yellow prussic acid, which is Fe 3+Therefore, Prussian blue can be formed by erythrocytes and Fe 3+ The reducing activity of a test sample can be measured using the following method.

[0086] 0.3 mL of sample (experimental group) or dilution water (blank group) was mixed with 0.3 mL of 0.2 M potassium phosphate buffer (pH 6.6) and 0.3 mL of 1% aqueous solution of red ferric chloride to prepare a mixed solution, which was then placed in a water bath at 50°C for 20 minutes. After the mixture was cooled to room temperature, a 10% aqueous solution of trichloroacetic acid was added to the mixed solution, which was then centrifuged at 3000 rpm for 20 minutes. 1.2 mL of the supernatant was taken, to which 1.2 mL of dilution water was added, and the mixture was uniformly mixed with 0.24 mL of 0.1% ferric chloride to prepare a reaction solution. The reaction solution was then left to stand for 10 minutes, and the OD 700 The absorbance value was detected to evaluate the amount of Prussian blue produced, which is used as an indicator of reducing activity.

[0087] The reducing activity was measured by the OD of the sample. 700 OD of the blank group from the readings 700 The temperature was set to 0.05°C minus the reading of 0.05°C.

[0088] 5.Skin irritation analysis

[0089] Skin irritation analysis of the test samples was performed using keratinocytes according to OECD 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method).

[0090] EpiDerm® bionic skin was placed in assay medium (EPI-100-NMM) and incubated at 37° C., 5% CO 2 The bioengineered skin was then transferred to fresh assay medium and incubated at 37° C., 5% CO 2The bioengineered skin was then incubated overnight (18±3 hours) at 37° C. in a cell incubator. Test substances (25 mg of solid or 30 μL of liquid) were added to the bioengineered skin, and the bioengineered skin was then incubated at 37° C. in 5% CO. 2 The bioengineered skin was placed in a cell incubator and incubated at 37 °C for 35 ± 1 min. After about 35 ± 1 min, the bioengineered skin was removed and placed on a laminar flow bench with the entire test material in contact with the bioengineered skin for 60 ± 1 min. The test material was then washed off the bioengineered skin with DPBS.

[0091] The bioengineered skin described above was then placed in the assay medium and incubated at 37°C, 5% CO 2 After incubation for 24 ± 2 h in a cell incubator at 37 °C and 5% CO, the assay medium was replaced and the bioengineered skin was incubated at 37 °C and 5% CO. 2 The plates were then placed back in the cell incubator and incubated for 18±2 hours.

[0092] The bioengineered skin was then incubated at 37°C, 5% CO 2 After incubation with 1 mg / mL MTT solution for 3 hours ± 5 minutes in a cell incubator, the bioengineered skin was washed with DPBS.

[0093] Finally, the purple crystals were dissolved in isopropanol and OD 570 The survival rate was calculated using the readings. If the survival rate was ≦50%, the test substance was considered to be a skin irritant.

[0094] 6.Skin sensitization analysis

[0095] Skin sensitization analysis of the test samples was performed using keratinocytes according to OECD442D (In Vitro Skin Sensitisation-ARE-Nrf2 Luciferase Test Method).

[0096] The test substances were diluted 2-fold from the highest concentration of 400 μg / mL with 1% DMSO aqueous solution. Then, different concentrations of the test substances were added to KeratinoSens® cells and incubated at 37° C., 5% CO2 The cells were incubated in a cell incubator at 37 °C for 48 hours.

[0097] The cells were then analyzed for intracellular Nrf2 activity using the Luciferase Assay System (Promega, E1501), and cell viability was detected using the MTT method. If the ratio of Nrf2 induction was greater than 1.5-fold when cell viability was greater than 70%, the test substance was considered to have skin sensitization properties.

[0098] 7.Eye irritation evaluation

[0099] The eye irritation analysis of the test samples was performed using rabbit corneal cells according to OECD491 (Short Time Exposure In Vitro Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Classification of Chemicals Not Requiring for Eye Irritation or Serious Eye Damage).

[0100] The test samples were adjusted to concentrations of 5% and 0.05% with physiological saline, and then SIRC cells were added. After standing at room temperature for 5 min, the cells were washed twice with PBS. The cells were then added to a medium containing 0.5 mg / mL MTT and incubated at 37°C, 5% CO 2 The cells were cultured in a cell incubator at 37 °C for 2 hours.

[0101] Finally, the purple crystals were dissolved in 0.04 N hydrochloric acid-isopropanol and OD 570The survival rate was calculated using the readings from the 0.5% and 0.05% test samples. If the survival rate was greater than 70% for the 0.5% and 0.05% test samples, the test samples were considered to belong to No Category (not required to be classified as eye irritants or serious eye damage). If the survival rate was 70% or less for the 0.5% and 0.05% test samples, the test samples were considered to belong to Category 1 (causing serious eye damage).

[0102] 8. Stability analysis

[0103] Test samples were treated with or without preservatives and were sampled at various time points to assay enzyme activity and confirm stability.

[0104] The fermentation stock solution prepared by the above-mentioned method was used as the test sample. The fermentation stock solution with each preservative was used as three experimental groups, while the fermentation stock solution without preservative was used as the control group. The preservatives used in the three experimental groups were 0.3% chlorphenesin, 1% para-hydroxyacetophenone, and a mixture of 0.2% chlorphenesin and 0.6% para-hydroxyacetophenone, respectively. The three experimental groups and the control group were aseptically packaged and stored at 25°C, and then sampled at specific time points to analyze the enzyme activity according to the above-mentioned method. The specific time points were the day of packaging, and 1, 2, 3, 7, 10, 14, 17, 21, 28, 35, 42, 49 and 56 days after packaging.

[0105] 9. Preparation of plant extracts as alkaline agents

[0106] The plant or plant-derived substrate was ground to obtain a powder. The powder was mixed with water in a weight ratio of 1:10, and extracted by ultrasonic treatment for 1 hour to obtain a crude extract. The crude extract was then filtered through a 0.8 μm membrane to remove excess impurities, and the obtained filtrate was used as the plant-derived alkaline agent.

[0107] The plant-derived alkaline agent may be further concentrated at 50° C. under reduced pressure.

[0108] B. Experimental Results

[0109] 1. Example 1

[0110] Analysis of fermentation stock

[0111] (1) Optimization of enzyme reaction conditions for fermentation liquid

[0112] (i) Reaction temperature

[0113] The fermentation liquid produced by Bacillus licheniformis GI_5E-1 was subjected to the enzyme activity analysis described in the experimental method above, and the enzyme activity was measured at 10°C intervals from 30°C to 80°C.

[0114] The results are shown in FIG.

[0115] As can be seen from Figure 1, the fermentation liquid produced by Bacillus licheniformis GI_5E-1 had excellent enzyme activity at all temperatures, with the enzyme activity being the highest at 60°C (OD 440 =1.9 (enzyme content 939U / mL)).

[0116] (ii) Reaction pH

[0117] The reaction pH of the fermentation stock solution produced by Bacillus licheniformis GI_5E-1 was adjusted from pH 4 to pH 11 by adjusting the pH value of 50 mM phosphate buffer used to prepare 0.5% azocasein solution according to the experimental method described above, and the enzyme activity was measured at each pH value.

[0118] The results are shown in FIG.

[0119] As can be seen from Figure 2, the fermentation liquid produced by Bacillus licheniformis GI_5E-1 had excellent enzyme activity at each pH, ​​and had even better enzyme activity in an alkaline environment (pH 8 to pH 11).

[0120] (2) Efficiency of disulfide bond cleavage in fermentation liquid

[0121] The fermentation liquid produced by Bacillus licheniformis GI_5E-1 was subjected to the analysis of disulfide bond cleavage efficiency as described in the above-mentioned experimental method. The reaction temperature was set to 60℃ or 90℃, and the disulfide bond cleavage efficiency of the fermentation liquid was analyzed.

[0122] The results are shown in FIG.

[0123] 3, it can be seen that the fermentation stock solution had excellent enzyme activity at all temperatures and had the ability to cleave disulfide bonds at both 60° C. and 90° C. In addition, the disulfide bond cleavage efficiency of the fermentation stock solution was superior at 60° C. compared to that at 90° C. (3.43% vs. 12.41%).

[0124] (3) Analysis of disulfide bond cleavage efficiency and reducing activity of fermentation stock solutions produced by different strains

[0125] The disulfide bond cleavage efficiency was analyzed as described in the experimental method above for the fermentation liquid produced from commercially available Bacillus licheniformis BCRC 14353 or the fermentation liquid produced from Bacillus licheniformis GI_5E-1. The reaction temperature was adjusted from 60°C to 30°C or 50°C to evaluate the disulfide bond cleavage efficiency of the fermentation liquid.

[0126] In addition, the fermentation stock solution produced by commercial Bacillus licheniformis BCRC 14353 or Bacillus licheniformis GI_5E-1 was subjected to the reducing activity analysis described in the experimental method described above. The reaction temperature was set at 30°C or 50°C to evaluate the reducing activity of the fermentation stock solution.

[0127] The results are shown in FIG.

[0128] As can be seen from Figure 4, the fermentation liquid produced from Bacillus licheniformis BCRC 14353 had disulfide bond cleavage ability and reducing activity at both 30°C and 50°C. Similarly, the fermentation liquid produced from Bacillus licheniformis GI_5E-1 also had disulfide bond cleavage ability and reducing activity at both 30°C and 50°C.

[0129] In addition, the disulfide bond cleavage efficiency of the fermentation liquid produced from Bacillus licheniformis GI_5E-1 was superior to that of the fermentation liquid produced from Bacillus licheniformis BCRC 14353 at both 30 and 50 °C.

[0130] (4) Skin irritation analysis of fermentation concentrate

[0131] The fermentation concentrate produced by Bacillus licheniformis GI_5E-1 was subjected to the skin irritation analysis described in the experimental method above.

[0132] The results are shown in FIG.

[0133] As can be seen from Figure 5, the cell viability of the stratum corneum cells treated with the fermentation concentrate produced by Bacillus licheniformis GI-5E-1 was 69.3%. In detail, according to the OECD439 guideline, a cell viability of more than 50% of the cells treated with the test sample means that the test sample is not skin irritating. Therefore, the fermentation concentrate produced by Bacillus licheniformis GI_5E-1 is considered to be non-skin irritating.

[0134] (5) Skin sensitization analysis of fermentation concentrate

[0135] The fermentation concentrate produced by Bacillus licheniformis GI_5E-1 was diluted to prepare test solutions of different concentrations, and the test solutions of different concentrations were subjected to the skin sensitization analysis described in the experimental method above.

[0136] The results are shown in FIG.

[0137] FIG. 6 shows that the cell viability of the stratum corneum cells treated with the test solutions of different concentrations was higher than 70%, and the allergen-inducing factors produced by the stratum corneum cells treated with the test solutions of different concentrations were all less than 1.5.

[0138] In detail, according to the OECD442D guideline, if the cell viability of the cells treated with the test sample is higher than 70% or the allergen inducer factor generated by the cells treated with the test sample is less than 1.5, it means that the test sample has no skin sensitization potential. Therefore, the fermentation concentrate produced by Bacillus licheniformis GI_5E-1 is considered to have no skin sensitization potential.

[0139] (6) Eye irritation analysis of fermentation concentrate

[0140] The fermentation concentrate produced by Bacillus licheniformis GI_5E-1 was diluted to prepare test solutions with different enzyme activities, and the test solutions with different enzyme activities were subjected to the eye irritation analysis described in the experimental method above.

[0141] The results are shown in FIG.

[0142] As can be seen from Figure 7, the cell viability of rabbit corneal cells treated with the test solutions with enzyme activity of 3,000 U / mL or less was higher than 70%. In detail, according to the OECD491 guideline, if the cell viability of the cells treated with the test sample is higher than 70%, it means that the test sample is not irritating to the eye. Therefore, when the fermentation stock solution produced by Bacillus licheniformis GI_5E-1 is diluted to an enzyme activity of 3,000 U / mL or less, it is considered to be not irritating to the eye.

[0143] (7) Stability analysis of fermentation liquid

[0144] The fermentation concentrate produced by Bacillus licheniformis GI_5E-1 was subjected to stability analysis as described in the experimental method above.

[0145] The results are shown in FIG.

[0146] As can be seen from Figure 8, the fermentation liquid produced by Bacillus licheniformis GI-5E-1 can maintain good enzyme activity for 55 days with or without the addition of preservatives, and the enzyme activity does not decrease significantly. In other words, the fermentation liquid produced by Bacillus licheniformis GI_5E-1 can maintain excellent stability even without the addition of preservatives.

[0147] Example 2

[0148] Analysis of polymer separation liquid

[0149] The polymer fraction of the fermentation broth produced by Bacillus licheniformis GI_5E-1 was subjected to electrophoretic analysis.

[0150] The results are shown in FIG.

[0151] As can be seen from FIG. 9, the polymer separation solution of the fermentation raw liquid produced by Bacillus licheniformis GI_5E-1 contains a protein with a molecular weight of 63 kDa.

[0152] After the proteins were extracted from the electrophoresis gel, protein identification was commissioned to ENERGENESIS BIOMEDICAL CO., LTD. using Proteome Discoverer V2.2.0.388 software.

[0153] The results are shown in Table 3.

[0154] [Table 3]

[0155] From Table 3, it can be seen that the protein with a molecular weight of 63 kDa contained in the polymeric separation solution of the fermentation raw liquid produced from Bacillus licheniformis GI_5E-1 may be an alkaline protease or keratinase with a higher identification score.

[0156] Example 3

[0157] Analysis of small molecule isolates with improved potency

[0158] 1. Identification of the components of the small molecule separation solution with improved efficacy

[0159] ENERGENESIS BIOMEDICAL CO., LTD. was commissioned to identify peptides in the components of the small molecule isolated solution produced by Bacillus licheniformis GI_5E-1, which is an effective fermentation concentrate, using Proteome Discoverer V2.2.0.388 software.

[0160] The identification results are shown in Table 4.

[0161] [Table 4]

[0162] From Table 4, it can be seen that there are 10 peptides with different sequences in the small molecule separation solution.

[0163] 2. Peptide Reductive Activity Analysis

[0164] The synthesis of the above 10 peptides was entrusted to ENERGENESIS BIOMEDICAL CO., LTD. The 10 synthesized peptides were then subjected to the reducing activity assay described in the above-mentioned experimental method to evaluate their reducing activity.

[0165] The results are shown in FIG.

[0166] As can be seen from FIG. 10, all of the above 10 peptides have reducing activity, among which peptide 2, peptide 3, peptide 6 and peptide 10 have higher reducing activity.

[0167] Example 4

[0168] Analysis of plant-derived extracts as alkaline agents

[0169] 1. Analysis of disulfide bond cleavage efficiency of fermentation stock in combination with different plant-derived extracts

[0170] The tea stems, mulberry leaves and coffee grounds were subjected to the preparation of plant-derived extracts as alkaline agents as described in the experimental method above, to obtain an aqueous extract of tea stems (containing theophylline), an aqueous extract of mulberry leaves (containing plant alkaloids) and an aqueous extract of coffee grounds (containing nicotinic acid).

[0171] The above extract was mixed with the fermentation liquid produced by Bacillus licheniformis GI_5E-1 in a volume ratio of 1:1 to prepare a mixture, which was then subjected to the disulfide bond cleavage efficiency analysis as described in the experimental method above.

[0172] The results are shown in FIG.

[0173] As can be seen from Figure 11, after the fermentation liquid is mixed with the above-mentioned various plant-derived extracts, the disulfide bond cleavage efficiency of the fermentation liquid is significantly improved. Among them, the use of the aqueous extract of coffee grounds as an alkaline agent assists the fermentation liquid, and the disulfide bond cleavage efficiency of the fermentation liquid is increased by 10 times. In addition, the aqueous extract of coffee grounds can absorb the odor peculiar to the fermentation liquid.

[0174] 2. Effect of alkaline agent concentration on disulfide bond cleavage efficiency in fermentation stock solution

[0175] The aqueous extract of the aforementioned coffee grounds was used as an alkaline agent and added to the fermentation stock solution produced by Bacillus licheniformis GI_5E-1 at concentrations of 10% (v / v), 30% (v / v), or 50% (v / v) to prepare mixtures, which were then subjected to the disulfide bond cleavage efficiency analysis as described in the experimental method above.

[0176] The results are shown in FIG.

[0177] As can be seen from FIG. 12, when the alkaline agent concentration was 30%, the disulfide bond cleavage efficiency of the fermentation concentrate was the highest at 11.2%, and compared to when the alkaline agent concentration was 10%, the disulfide bond cleavage efficiency of the fermentation concentrate at an alkaline agent concentration of 30% was improved by approximately 26%.

[0178] Example 5

[0179] Analysis of disulfide bond cleavage efficiency of hair shape altering compositions with different formulations

[0180] The polymeric isolated solution and the enhanced small molecule isolated solution of the fermentation liquid produced by Bacillus licheniformis GI_5E-1 were diluted to adjust the enzyme activity and reducing power, respectively. Then, each polymeric isolated solution and each enhanced small molecule isolated solution were combined and mixed with 30% (v / v) water extract of coffee grounds in a volume ratio of 1:1 to prepare five hair shape modification composition samples with different formulations as shown below.

[0181] (i) Sample S1: containing 33.3% (v / v) of polymeric isolated solution, 11% (v / v) of small molecule isolated solution, 15% (v / v) of water extract of coffee grounds, and 40.7% (v / v) of pure water.

[0182] (ii) Sample S2: containing 21.5% (v / v) of polymeric isolation solution, 9% (v / v) of small molecule isolation solution, 15% (v / v) of water extract of coffee grounds, and 54.5% (v / v) of pure water.

[0183] (iii) Sample S3: containing 21.5% (v / v) of polymeric isolate, 14% (v / v) of small molecule isolate, 15% (v / v) of water extract of coffee grounds, and 49.5% (v / v) of pure water.

[0184] (iv) Sample S4: containing 21.5% (v / v) of polymeric isolation solution, 18% (v / v) of small molecule isolation solution, 15% (v / v) of water extract of coffee grounds, and 45.5% (v / v) of pure water.

[0185] (v) Sample S5: containing 21.5% (v / v) of polymeric isolation solution, 21% (v / v) of small molecule isolation solution, 15% (v / v) of water extract of coffee grounds, and 42.5% (v / v) of pure water.

[0186] Five samples were subjected to the analysis of disulfide bond cleavage efficiency as described in the Experimental Methods above.

[0187] The results are shown in Figure 13.

[0188] As can be seen from FIG. 13, all of the five samples described above have excellent disulfide bond cleavage efficiency, and among them, sample S5 has the highest disulfide bond cleavage efficiency of 16.4%.

[0189] It will be apparent to those skilled in the art that various modifications and variations can be made to the disclosed embodiments. It is intended that the specification and examples be considered as exemplary only, with the true scope of the present disclosure being indicated by the following claims and their equivalents.

Claims

1. a polymer component comprising a protease belonging to the alkaline protease class; a small molecule component comprising a peptide having reducing activity; An alkaline agent component; A biological hair shape modifying composition comprising: The protease belonging to the alkaline protease class includes alkaline protease, keratinase, or a combination thereof; The peptides having reducing activity include at least one of the following peptides: (b) a peptide comprising the amino acid sequence SEQ ID NO. 2; (c) a peptide comprising the amino acid sequence SEQ ID NO. 3; (f) a peptide comprising the amino acid sequence SEQ ID NO. 6; and (j) a peptide comprising the amino acid sequence SEQ ID NO. 10; both the macromolecular and small molecule components are derived from the fermentation product of Bacillus licheniformis and a medium containing keratin and / or keratin polymers; The molecular weight of the component in the polymer component is 3 kDa or more and 3 to 1000 kDa; A biological hair shape modifying composition, wherein the molecular weight of the ingredients in said small molecule component is less than 3 kDa and is between 0.01 and 2.99 kDa.

2. The biological hair shape modifying composition of claim 1 , wherein the alkaline agent component comprises a plant-derived alkaline agent and / or a chemically synthesized alkaline agent.

3. 3. The biological hair shape modifying composition of claim 1 or 2, wherein the concentration of the alkaline agent component in the biological hair shape modifying composition is 1-80% (v / v).

4. Regarding the Bacillus licheniformis 3. The biological hair shape modifying composition of claim 1 or 2, wherein the Bacillus licheniformis comprises Bacillus licheniformis BCRC 14353, Bacillus licheniformis GI_5E-1 with deposit number BCRC 911027, or a combination thereof.

5. The biological hair shape modifying composition of claim 1 or 2, wherein the keratin and / or keratin polymer-containing medium comprises a keratin and / or keratin polymer-containing substrate, and the keratin and / or keratin polymer-containing substrate comprises animal feathers, hair, hooves, claws and / or horns.

6. 3. The biological hair shape modifying composition of claim 1 or 2, wherein the volume ratio of said polymer component to said small molecule component in said hair shape modifying composition is 5-10:1.5-3.

7. A biological hair shape modification kit, comprising: a polymer component comprising a protease belonging to the alkaline protease class; a small molecule component comprising a peptide having reducing activity; An alkaline agent component; at least one packaging container for containing the polymer component, the small molecule component and the alkaline agent component; Including, The protease belonging to the alkaline protease class includes alkaline protease, keratinase, or a combination thereof; The peptides having reducing activity include at least one of the following peptides: (b) a peptide comprising the amino acid sequence SEQ ID NO. 2; (c) a peptide comprising the amino acid sequence SEQ ID NO. 3; (f) a peptide comprising the amino acid sequence SEQ ID NO. 6; and (j) a peptide comprising the amino acid sequence SEQ ID NO. 10; both the macromolecular and small molecule components are derived from the fermentation product of Bacillus licheniformis and a medium containing keratin and / or keratin polymers; The molecular weight of the component in the polymer component is 3 kDa or more and is 3 to 1000 kDa, and the molecular weight of the component in the small molecule component is less than 3 kDa and is 0.01 to 2.99 kDa; The biological hair shape modification kit is used in the form of a mixture of the polymer component, the small molecule component and the alkaline agent component.

8. The biological hair shape modification kit according to claim 7 , wherein the alkaline agent component includes a plant-derived alkaline agent and / or a chemically synthesized alkaline agent.

9. The biological hair shape modification kit according to claim 8, wherein the plant-derived alkaline agent includes at least one of the following plant-derived alkaline agents: an aqueous extract of coffee grounds, an aqueous extract of tea stems, and an aqueous extract of mulberry leaves.

10. The biological hair shape modification kit according to any one of claims 7 to 9, wherein the Bacillus licheniformis includes Bacillus licheniformis BCRC 14353, Bacillus licheniformis GI_5E-1 with deposit number BCRC 911027, or a combination thereof.

11. The biological hair shape modification kit according to any one of claims 7 to 9, wherein the keratin and / or keratin polymer-containing medium comprises a keratin and / or keratin polymer-containing substrate, and the keratin and / or keratin polymer-containing substrate comprises animal feathers, hair, hooves, claws and / or horns.

12. The biological hair shape modification kit according to claim 11, wherein the content of the keratin and / or keratin polymer-containing substrate in the keratin and / or keratin polymer-containing medium is 0.01 to 20 g / L.

13. In the biological hair shape modification kit, the macromolecular component is contained in a first reagent, the small molecule component is contained in a second reagent, and the alkaline agent component is contained in a third reagent; The at least one packaging container includes a first container, a second container, and a third container, the first reagent is packaged in the first container, the second reagent is packaged in the second container, and the third reagent is packaged in the third container; The biological hair shape modification kit is used by mixing the first reagent, the second reagent, and the third reagent. The biological hair shape modification kit according to any one of claims 7 to 9.

14. The biological hair shape modification kit is used by mixing the first reagent, the second reagent, the third reagent and additional water in a volume ratio of 5 to 10: 1.5 to 3: 3 to 6: 1 to 10.

5. The biological hair shape modification kit according to claim 13.

15. The biological hair shape altering kit is used by pouring out all of the first reagent, the second reagent, and the third reagent from the first container, the second container, and the third container, respectively, and mixing them with additional water. The biological hair shape modification kit according to claim 13.

16. In the biological hair shape modification kit, the polymer component and the small molecule component are contained in a first reagent, and the alkaline agent component is contained in a second reagent; the at least one packaging container includes a first container and a second container, the first reagent is packaged in the first container, and the second reagent is packaged in the second container; The biological hair shape modification kit is used by mixing the first reagent and the second reagent. The biological hair shape modification kit according to any one of claims 7 to 9.

17. The biological hair shape modification kit according to claim 16, wherein the biological hair shape modification kit is used by mixing the first reagent, the second reagent, and additional water in a volume ratio of 6.5 to 13:3 to 6:1 to 10.

5.

18. 17. The biological hair shape altering kit of claim 16, wherein the biological hair shape altering kit is used by pouring out all of the first and second reagents from the first and second containers, respectively, and mixing them with additional water.

19. In the biological hair shape modification kit, the polymer component, the small molecule component and the alkaline agent component are contained in a first reagent, the at least one packaging container includes a first container, and the first reagent is packaged in the first container; The biological hair shape modification kit according to any one of claims 7 to 9.

20. A method for altering hair shape, comprising: (a) a hair permeation step of applying a mixture produced using the biological hair shape modification composition according to claim 1 or 2 or the biological hair shape modification kit according to any one of claims 7 to 9 to hair and permeating the hair; (b) a hair shape modification step of subjecting the hair to which the biological hair shape modification composition or the mixture has been applied and permeated, to a heating process to hydrolyze the protein backbone of the hair and cleave disulfide bonds in the protein of the hair, thereby modifying the shape of the hair; A method for altering hair shape comprising:

21. The method for changing the hair shape according to claim 20, wherein the temperature of the heating step is between 40 and 100°C.

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