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Method for reducing misincorporation of non-canonical branched-chain amino acids

A branched-chain amino acid, non-classical technology, applied in the field of reducing the mis-incorporation of non-classical branched-chain amino acids, can solve the problem of incomplete clarity of non-classical branched-chain amino acids and the like

Pending Publication Date: 2021-10-26
SANOFI AVENTIS DEUT GMBH +1
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  • Abstract
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  • Application Information

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Problems solved by technology

[0008] The conditions under which misincorporation of non-canonical branched-chain amino acids into heterologous recombinant proteins occur and how to effectively prevent such misincorporation are not fully understood

Method used

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  • Method for reducing misincorporation of non-canonical branched-chain amino acids
  • Method for reducing misincorporation of non-canonical branched-chain amino acids
  • Method for reducing misincorporation of non-canonical branched-chain amino acids

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Embodiment Construction

[0027] As noted above, the present invention relates to methods for producing recombinant polypeptides of interest in microbial host cells. The method comprises (a) introducing into a microbial host cell a polynucleotide encoding the polypeptide of interest that has been modified such that an enzyme activity as described elsewhere herein in the microbial host cell is compatible with said enzymatic activity is modulated in an unmodified microbial host cell; and (b) expressing said polypeptide of interest in said microbial host cell.

[0028] According to the present invention, the recombinant polypeptide of interest should be produced in a microbial host cell. The term "recombinant polypeptide" as used herein refers to a genetically engineered polypeptide. Therefore, the polypeptide to be produced should be heterologous with respect to the microbial host cell, meaning that the host cell does not naturally express the polypeptide of interest. Thus, the term "heterologous" mean...

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Abstract

The present invention relates to a method for producing a recombinant polypeptide of interest in a microbial host cell, comprising (a) introducing a polynucleotide encoding the polypeptide of interest into a microbial host cell which has been modified such that an enzymatic activity selected from the group consisting of ketol-acid reductoisomerase (NADP(+)) activity (EC 1.1.1.86), acetohydroxyacid synthase activity (EC 2.2.1.6), aspartate kinase activity (EC 2.7.2.4), homoserine dehydrogenase activity (EC 1.1.1.3), and L-threonine dehydratase activity (EC 4.3.1.19) is modulated in said microbial host cell as compared to the enzymatic activity in an unmodified microbial host cell, and (b) expressing said polypeptide of interest in said microbial host cell. Moreover, the present invention relates to a method for reducing misincorporation of at least one non-canonical branched-chain amino acid into a recombinant polypeptide of interest expressed in a microbial host cell.

Description

technical field [0001] The present invention relates to a method for producing a target recombinant polypeptide in a microbial host cell, comprising (a) introducing into a microbial host cell a polynucleotide encoding the target polypeptide, the microbial host cell having been modified such that The microbial host cell is selected from ketol acid reductoisomerase (NADP (+)) activity (EC 1.1.1.86), acetohydroxyacid synthase activity (EC 2.2.1.6), aspartokinase activity ( EC 2.7.2.4), homoserine dehydrogenase activity (EC 1.1.1.3) and L-threonine dehydratase activity (EC 4.3.1.19) enzymatic activity compared with said enzymatic activity in unmodified microbial host cells modulated (eg, increased); and (b) expressing said polypeptide of interest in said microbial host cell. Furthermore, the present invention relates to a method for reducing misincorporation of at least one non-canonical branched chain amino acid into a recombinant polypeptide of interest expressed in a microbial...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/00C12N9/04C12N9/10C12N9/88
CPCC07K14/62C12N9/0006C12N9/1022C12N9/1217C12N9/88C12Y101/01003C12Y101/01086C12Y202/01006C12Y207/02004C12Y403/01009C12P21/02C12P13/04C12P13/06C12P13/08C12N15/70C12Y403/01019
Inventor P·豪普特曼A·科尔科莱斯·加西亚C·T·拉特曼A·马特森P·诺伊鲍尔
Owner SANOFI AVENTIS DEUT GMBH
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