CN-122012472-A - Acid serine keratinase gene and application thereof

Abstract

The invention discloses an acidic serine keratinase gene and application thereof, and belongs to the technical field of genetic engineering. The novel serine keratinase is obtained by screening from the Yunnan cattle street acidic hot spring environment, has the characteristics of acid resistance, good thermal stability and high enzyme activity, can be well adapted to the physiological acidic microenvironment of skin and the whole process temperature fluctuation of product production, storage and transportation, does not need complex stabilization treatment, can simplify the production process, reduce the production cost, prolong the shelf life of the product, realize green and efficient preparation, solves the key problems of irritation, poor stability, difficult storage and the like of the traditional keratinase in skin application from the source, is particularly suitable for the mild peeling of human corner plugs, and has outstanding application value and broad prospect in the daily chemical skin care field.

Inventors

• ZHANG YANYAN
• PENG BINGBING
• ZOU ZHIRONG
• GUO LING
• Xiao Rongshen
• CHEN YONGLI
• TANG LIFENG
• ZHANG YANFENG
• GU LIHONG
• Zhou Huwu
• LI YAN
• ZHANG KAIMIN

Assignees

• 深圳中科欣扬生物科技有限公司
• 广州稀咖科技有限公司

Dates

Publication Date

20260512

Application Date

20260410

Claims (10)

1. 1. Serine keratinase TVG06-16620, characterized in that the amino acid sequence is shown in SEQ ID NO. 1.
2. 2. A gene encoding the serine keratinase TVG06-16620 of claim 1.
3. 3. A recombinant vector comprising the gene according to claim 2.
4. 4. Recombinant strain, characterized in that it expresses the serine keratinase TVG06-16620 of claim 1 or contains the recombinant vector of claim 3.
5. 5. The recombinant strain of claim 4, wherein the host cell includes, but is not limited to, E.coli, C.glutamicum, B.subtilis, A.lactis, or a yeast.
6. 6. Recombinant escherichia coli, characterized in that the serine keratinase TVG06-16620 of claim 1 is expressed by using escherichia coli BL21 (DE 3) as a host and pET-28a as a vector.
7. 7. A method for preparing the serine keratinase TVG06-16620 of claim 1, characterized in that the recombinant strain of claim 4 or 5 or the recombinant escherichia coli of claim 6 is fermented to induce the expression of the serine keratinase TVG06-16620 and the serine keratinase TVG06-16620 is collected.
8. 8. Use of serine keratinase TVG06-16620 of claim 1 for the preparation of a product useful for the exfoliation and/or dissolution of keratin plugs.
9. 9. The use according to claim 8, wherein the product is a daily chemical product, including but not limited to a rinse-off cleansing product or a leave-on skin care product.
10. 10. The use according to claim 9, wherein the rinse-off cleansing product comprises a cleansing cream, a body wash, a body cream or a exfoliating cream, and the leave-on skin care product comprises a lotion, an essence or a mask.

Description

Acid serine keratinase gene and application thereof Technical Field The invention relates to a slave acid serine keratinase gene and application thereof, belonging to the technical field of genetic engineering. Background Serine keratinase (Serine Proteases) contains serine (Ser), histidine (His) and aspartic acid (Asp) catalytic triplets in the active center, and is the most widely used class of keratinase family. From the source, the enzyme is mainly separated from microbial strains such as bacillus subtilis, beauveria bassiana, bacillus licheniformis and the like, has disulfide bond reduction and polypeptide hydrolysis activities, can efficiently break down compact bonding networks in keratin molecules, degrades insoluble keratin into soluble polypeptides or amino acids, has little damage to collagen and elastin, and has natural biocatalysis advantages. Compared with animal and plant source enzymes, the microbial source serine keratinase is easy to be industrially produced through fermentation, and the enzymatic properties can be optimized through genetic engineering. In daily chemical care, the main flow of keratin treatment and care components is two types, namely chemical stripping, acid, salicylic acid and derivatives thereof, wherein the substances have strong irritation, skin barrier damage is easily caused by long-term use, a large amount of energy sources are consumed in the chemical synthesis process, high pollution is produced, green production cannot be realized, biological enzymes have the mildness remarkably better than chemical types, the enzymatic stability is poor, the skin is easily inactivated due to the influence of pH value and temperature, and the enzyme activity is greatly attenuated in an acidic environment with the pH value of 4.5-6.0 on the surface layer of the skin, and the inactivation is caused by temperature fluctuation in production, storage and use. Serine keratinase from natural sources is fresh and difficult to meet the comprehensive performance requirements of acid resistance, high temperature resistance and high enzyme activity at the same time. Therefore, it is important to screen keratinase that is acid-resistant, high temperature resistant and has high catalytic activity. Although some new keratinase enzymes are provided in the prior art, such as the acid keratinase enzyme obtained by screening of patent CN117286126A, the acid keratinase enzyme is only applicable to the wool processing field, is difficult to apply to the skin environment and has limited application scenes. Disclosure of Invention Aiming at the prior art, the invention provides an acidic thermostable serine keratinase TVG06-16620 screened from Yunnan cattle street acidic hot spring and a coding gene thereof, an expression vector and a recombinant strain comprising the gene, and a hornbolt stripping effect of the enzyme. The novel serine keratinase is screened from Yunnan cattle street acid hot spring, is connected with an expression vector pET-28a, is transformed into escherichia coli BL21 (DE 3) for induction expression, researches the enzymology property of the novel serine keratinase, and realizes the optimal dosage of the enzyme for dissolving human angular thrombus through formula optimization. The invention has important significance for understanding the characteristics and efficacy of the acid thermostable serine keratinase TVG06-16620 from microbial sources. The invention is realized by the following technical scheme: the invention provides serine keratinase TVG06-16620, the amino acid sequence of which is shown in SEQ ID NO. 1. The invention also provides a gene encoding serine keratinase TVG 06-16620. In one embodiment, the nucleotide sequence of the gene is shown in SEQ ID NO. 2. The invention also provides a recombinant vector containing a gene for coding the acid thermostable serine keratinase TVG06-16620 and a complete coding reading frame sequence. In one embodiment, the vector is preferably pET-28a. The invention also provides a recombinant strain which expresses the serine keratinase TVG06-16620 or contains the recombinant vector. In one embodiment, the host cell of the recombinant strain includes, but is not limited to, E.coli, C.glutamicum, B.subtilis, L.lactis, or yeast. In one embodiment, the recombinant strain is hosted by E.coli ESCHERICHIA COLIBL (DE 3). In one embodiment, the recombinant strain contains the recombinant vector, and the gene encoding the acid thermostable serine keratinase TVG06-16620 is sufficiently expressed in a recipient cell through a promoter operation, thereby conferring the recipient bacterium the ability to produce the acid thermostable serine keratinase TVG 06-16620. The invention also provides a method for preparing the serine keratinase TVG06-16620, the recombinant strain is fermented, the expression of the serine keratinase TVG06-16620 is induced, and the serine keratinase TVG06-16620 is collected. In one embodiment, the method further comprises co