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The patent badge is an abbreviated version of the USPTO patent document. The patent badge does contain a link to the full patent document.

The patent badge is an abbreviated version of the USPTO patent document. The patent badge covers the following: Patent number, Date patent was issued, Date patent was filed, Title of the patent, Applicant, Inventor, Assignee, Attorney firm, Primary examiner, Assistant examiner, CPCs, and Abstract. The patent badge does contain a link to the full patent document (in Adobe Acrobat format, aka pdf). To download or print any patent click here.

Patent No.:

US 12152255 B1

PDFFull Text
Date of Patent:
Nov. 26, 2024

Filed:

Dec. 06, 2021

Ω-transaminase mutant obtained by dna synthetic shuffling combined mutation and use

Applicants:

Zhejiang University of Science & Technology, Hangzhou, CN;

Enzymaster (Ningbo) Bio-engineering Co., Ltd., Ningbo, CN;

Inventors:

Jun Huang, Hangzhou, CN;

Chunyan Liu, Hangzhou, CN;

Lehe Mei, Hangzhou, CN;

Haibin Chen, Ningbo, CN;

Changjiang Lv, Hangzhou, CN;

Sheng Hu, Hangzhou, CN;

Hongpeng Wang, Hangzhou, CN;

Weirui Zhao, Hangzhou, CN;

Fangfang Fan, Hangzhou, CN;

Ye Li, Hangzhou, CN;

Linka Yu, Hangzhou, CN;

Yifeng Zhou, Hangzhou, CN;

Assignees:

ZHEJIANG UNIVERSITY OF SCIENCE & TECHNOLOGY, Hangzhou, CN;

ENZYMASTER (NINGBO) BIO-ENGINEERING CO., LTD., Ningbo, CN;

Attorneys:

Jiwen Chen

Joywin IP Law PLLC

Primary Examiner:

Yong D Pak

Int. Cl.
CPC ...
C12N 9/10 (2006.01); C12N 15/63 (2006.01); C12P 7/26 (2006.01); C12N 15/10 (2006.01); C12R 1/19 (2006.01); C12R 1/66 (2006.01);
U.S. Cl.
CPC ...
C12N 9/1096 (2013.01); C12N 15/63 (2013.01); C12P 7/26 (2013.01); C12N 15/1027 (2013.01); C12R 2001/19 (2021.05); C12R 2001/66 (2021.05); C12Y 206/01018 (2013.01);
Abstract

The present invention discloses a ω-transaminase mutant obtained through DNA synthetic shuffling combined mutation. The ω-transaminase mutant is obtained through point mutation of a wild type ω-transaminase from. The amino acid sequence of the wild type ω-transaminase is shown in SEQ ID NO: 1. The mutation site of the ω-transaminase mutant is any one of: (1) F115L-H210N-M150C-M280C; (2) F115L-H210N; (3) F115L-H210N-E253A-I295V; (4) I77L-F115L-E133A-H210N-N245D; (5) I77L-Q97E-F115L-L118T-E253A-G292D; (6) I77L-E133A-N245D-G292D; and (7) H210N-N245D-E253A-G292D. According to the present invention, forward mutations obtained in the previous stage are randomly combined through a DNA synthetic shuffling combined mutation method. It is verified through experiments that this method can effectively improve the probability of forward mutation and increase experimental efficiency and feasibility, and is capable of obtaining mutant enzymes with thermodynamic stability remarkably superior to that of wild enzymes via screening.

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