Enzymaster (ningbo) Bio-engineering Co., Ltd.

Novo Nordisk A/s

Zhejiang University

Haibin Chen

Yong Koy Bong

Baoqin Cai

Qing Xu

Lei Sun

Chengxiao Zhang

Marco Bocola

Chuanyang Shang

Tianran Shen

Yingxin Zhang

Ameng Zhou

Jiadong Yang

Shumin Shi

Zhuhong Yang

Zhe Wang

Li Ping Yang

Jing Su

Zhe Sun

Sebastian Beck Joergensen

Jun Huang

Wei Yang

Ye Li

Changjiang Lv

Sheng Hu

Weirui Zhao

Hongpeng Wang

Fangfang Fan

Linka Yu

Lehe Mei

Yaoyao Ji

Juanjuan Wang

Chih-Chuan Chang

Zhaoqi Zhang

Zhenhua He

Wenhui Zheng

Dongxiao Hu

Ruimei Hong

Kuifang He

Yifeng Zhou

Zikun Wang

Qinli Peng

Chunyan Liu

Provided are efficient catalyst of engineered enzymes and an economical enzymatic reaction solution to solve the problems in the current production process of L-tyrosine and its derivatives. The method of the invention has the advantages of high product concentration, mild reaction conditions, simple purification process, simple operation, environmental friendliness, and easy industrial scale-up. Thus, it has good industrial application prospects.

Engineered polypeptides and its application in the synthesis of tyrosine or tyrosine derivatives

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.

ω-Transaminase mutant obtained by DNA synthetic shuffling combined mutation and use

Provided are amino acid sequences of ketoreductase polypeptides that are useful for asymmetrically synthesizing chiral alcohol compounds, its preparation process as well as reaction process under industrial-relevant conditions. Also provided are polynucleotide sequences encoding engineered ketoreductase polypeptides, engineered host cells capable of expressing engineered ketoreductase polypeptides, and methods of producing chiral alcohol compounds using engineered ketoreductase polypeptides. Compared to other enzymes, the engineered ketoreductase polypeptides provided by the invention have better catalytic activity and thermal stability, allowing purification of the enzyme solution by heat treatment, which is advantageous for the production of enzymes and the industrial application of enzymatic reactions. The use of the engineered polypeptides of the present invention greatly simplifies the production process of chiral alcohol compounds, reduces the cost of production and the impact on the environment, and has good industrial application prospects.

Engineered ketoreductase polypeptides and uses thereof

Provided are amino acid sequences of engineered transaminase polynucleotide that are useful for asymmetrically synthesizing chiral amine compounds, and its preparation process and reaction process under industrial-relevant conditions. Also provided are polynucleotide sequences encoding engineered transaminase polypeptides, engineered host cells capable of expressing engineered transaminase polypeptides, and methods of producing chiral amine compounds using engineered transaminase polypeptides. Compared to other enzymes, the engineered transaminase polypeptides provided by this invention have better catalytic activity and stability, and are not inhibited by chiral amine products in the synthesis process. The use of the engineered polypeptides of the present invention for the preparation of chiral amine compound results in higher unit activity, and has good industrial application prospects.

Engineered transaminase polypeptides and uses thereof

Provided herein are engineered decarboxylase polypeptides that are useful for catalyzing the decarboxylation of amino acids such as L-tyrosine to produce tyramine or catalyzing the decarboxylation of L-DOPA to produce dopamine. Also provided are the preparation process of engineered decarboxylase polypeptides as well as reaction process under industrial-relevant conditions. The disclosure also provides polynucleotide sequences encoding engineered decarboxylase polypeptides, recombinant host cells capable of expressing engineered decarboxylase polypeptides, and methods of producing tyramine or dopamine using the engineered decarboxylase polypeptides. Compared to the wild type decarboxylase, the engineered polypeptide provided by this disclosure has better activity and/or stability. The use of the engineered polypeptides for the preparation of tyramine or dopamine reduces the production cost and has a good industrial application prospect.

Engineered decarboxylase polypeptides and their uses in preparing tyramine and dopamine

The present invention provides engineered polypeptides that are useful for the asymmetric synthesis of &#x3b2;-hydroxy-&#x3b1;-amino acids under industrial-relevant conditions. The engineered polypeptides disclosed in this invention were developed through directed evolution based on the ability of catalytic synthesis of (2S, 3R)-2-amino-3-hydroxy-3-(4-nitrophenyl) propanoic acid. The present disclosure also provides polynucleotides encoding engineered polypeptides, host cells capable of expressing engineered polypeptides, and methods of producing &#x3b2;-hydroxy-&#x3b1;-amino acids using engineered polypeptides. Compared to other processes of preparation, the use of the engineered polypeptides of the present invention for the preparation of &#x3b2;-hydroxy-&#x3b1;-amino acids results in high purity of the desired stereoisomers, mild reaction conditions, low pollution and low energy consumption. So, it has good industrial application prospects.

Engineered polypeptides and their applications in the synthesis of beta-hydroxy-alpha-amino acids

The present invention provides amino acid sequences of engineered decarboxylase polypeptides that are useful for catalyzing the decarboxylation of L-aspartate to produce &#x3b2;-alanine, and the preparation process of engineered decarboxylase polypeptides as well as reaction process under industrial-relevant conditions. The present disclosure also provides polynucleotide sequences encoding engineered decarboxylase polypeptides, engineered host cells capable of expressing engineered decarboxylase polypeptides, and methods of producing &#x3b2;-alanine using the engineered cells. Compared to the wild-type decarboxylase, the engineered decarboxylase polypeptide provided by the invention has better activity and stability, and overcomes the inhibition by L-aspartic acid and/or &#x3b2;-alanine. The use of the engineered polypeptides of the present invention for the preparation of &#x3b2;-alanine results in higher unit activity, lower cost, and has good industrial application prospects.

Engineered decarboxylase polypeptides and their application in preparing beta-alanine

The present invention provides engineered polypeptides that are useful for the asymmetric synthesis of &#x3b2;-hydroxy-&#x3b1;-amino acids under industrial-relevant conditions. The present disclosure also provides polynucleotides encoding engineered polypeptides, host cells capable of expressing engineered polypeptides, and methods of producing &#x3b2;-hydroxy-&#x3b1;-amino acids using engineered polypeptides. Compared to other processes of preparation, the use of the engineered polypeptides of the present invention for the preparation of &#x3b2;-hydroxy-&#x3b1;-amino acids results in high purity of the desired stereoisomers, mild reaction conditions, low pollution and low energy consumption. So, it has good industrial application prospects.

Engineered aldolase polypeptides and uses thereof

The present invention relates to the newly identified MIC-1 binding receptor, GFRAL. In vitro bioassays, for testing affinity and potency of GFRAL ligand, such as MIC- or MIC-1 variants, are provided.

MIC-1 receptor and uses thereof

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Ningbo, China

Haibin Chen