# US 12,595,440 B2 · 2709 alkali protease mutant modified based on molecular dynamics calculation and use thereof

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 12,595,440 B2
- Document ID: US12595440B2
- Status: Granted
- Kind: B2
- Application number: 18/021,627
- Filed: 2021-04-25
- Published: 2025-07-03
- Granted: 2026-04-07
- Earliest claimed: 2020-11-14
- Assignees: Shanxi Yong Forever Technology Co., Ltd
- Applicants: Shanxi Yong Forever Technology Co., Ltd
- Inventors: Jian Zhang
- Web page: https://patentgrep.com/patent/US12595440B2

## CPC

- C11D 3/386: Preparations containing enzymes {, e.g. protease or amylase}
- C11D 3/38618: {Protease or amylase in liquid compositions only}
- C12N 9/54: bacteria being Bacillus
- C12N 15/75: for Bacillus
- C12Y 304/21062: Subtilisin (3.4.21.62)
- C11D: DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Y: ENZYMES

## Abstract

The present disclosure belongs relates to a 2709 alkali protease mutant modified based on molecular dynamics calculation and use thereof in liquid detergents. The dynamic structure feature of a 2709 alkali protease derived from Bacillus licheniformis are studied by using a molecular dynamics method through computer simulation, which is a unique effective method capable of studying a relationship between the structure of the 2709 alkali protease and molecular dynamics as well as an enzyme function in an atomic level. After molecular dynamics calculation, appropriate sites are selected for protein engineering to obtain the 2709 alkali protease mutant suitable for industrial application of detergents. The protease mutant has significantly improved enzyme activity under the alkaline condition, is better in stability, not only has good decontamination effect but also has good component compatibility with components in the detergent, and can be widely applied to the field of washing industry.

## Claims

### Claim 1

A 2709 alkali protease mutant modified based on molecular dynamics calculation, wherein the parent protein of the alkali protease mutant is a 2709 alkali protease obtained from Bacillus licheniformis, the amino acid sequence of the alkali protease mutant SEQ ID NO: 2, SEQ ID NO:3 or SEQ ID NO:4.

### Claim 2 (depends on 1)

A plasmid comprising the polynucleotide sequence encoding the 2709 alkali protease mutant according to claim 1.

### Claim 3 (depends on 2)

A recombinant Bacillus licheniformis transformant comprising the plasmid according to claim 2.

### Claim 4 (depends on 1)

A detergent composition, comprising the 2709 alkali protease mutant according to claim 1.

### Claim 5 (depends on 1)

A method of use of a composition comprising 2709 alkali protease mutant according to claim 1 in the field of cleaning or washing.