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Promoters from corynebacterium glutamicum and uses thereof in regulating ancillary gene expression

A promoter and gene technology, applied in genetic engineering, virus/bacteriophage, using vectors to introduce foreign genetic material, etc., can solve the problems of undeveloped productivity and yield

Inactive Publication Date: 2020-03-06
ZYMERGEN INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in industrially important host strains, control over the productivity and yield of these biomolecules is achieved by operably linking a heterologous promoter to one or more accessory target genes, thereby regulating the expression of these target genes. Potential impact largely untapped

Method used

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  • Promoters from corynebacterium glutamicum and uses thereof in regulating ancillary gene expression
  • Promoters from corynebacterium glutamicum and uses thereof in regulating ancillary gene expression
  • Promoters from corynebacterium glutamicum and uses thereof in regulating ancillary gene expression

Examples

Experimental program
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Effect test

example 1

[0260] Example 1: Application of candidate promoters in the biosynthetic pathway of L-lysine

[0261] The promoters of the present disclosure can be used in improved methods for producing biomolecules in host cells. An example of the application and use of the promoter of the present disclosure involves the production of the amino acid L-lysine.

[0262] figure 1 A biosynthetic pathway for the production of L-lysine is presented, and contains the genes pck, odx, icd, and hom (for example, homoserine / threonine synthetase pathway), which convert intermediates from the pathway, This leads to a decrease in total L-lysine production. Table 3 provides the symbol, gene name, Enzymology Committee number (EC number) and map position in Corynebacterium glutamicum strain ATCC 13032.

[0263] Using yeast homologous recombination cloning technology, the recombinant vector including the promoters of SEQ ID NO: 1 to 8 functionally linked to the target gene provided in Table 3 was cloned into the...

example 2

[0276] Example 2: Engineering the biosynthetic pathway of L-lysine

[0277] The production of L-lysine is improved by swapping the promoter pairs of target genes. The construct of Example 1 was used to prepare the following recombinant organisms:

[0278] The combination of Pcg0007-lysA and Pcg3121-pgi provides the highest yield of L-lysine.

[0279] Table 7: Paired promoter exchanges of target genes in the L-lysine biosynthetic pathway

[0280]

[0281]

example 3

[0282] Example 3: Engineering the L-lysine biosynthetic pathway with a promoter operably linked to genes outside the pathway

[0283] The production of L-lysine is modified by including a second promoter polynucleotide sequence that is functionally linked to a second heterologous target gene outside the pathway. Heterologous target genes are selected from the group consisting of ncgl0009, ncgl0019, ncgl0054, ncgl0082, ncgl0142, ncgl0223, ncgl0241, ncgl0242, ncgl0304, ncgl0306, ncgl0356, ncgl0398, ncgl0408, ncgl0424, ncgl0425, ncgl0427, ncgl0439, ncgl0458, ncgl0439, ncgl0458, ncgl0439, ncgl0458, 31ncgl0564 ncgl0573, ncgl0578, ncgl0581, ncgl0598, ncgl0600, ncgl0601, ncgl0641, ncgl0663, ncgl0668, ncgl0737, ncgl0767, ncgl0813, ncgl0823, ncgl0827, ncgl0853, ncgl0874, ncgl0877, ncgl0905, ncgl1,187,ncglncglncgl1137, ncgl0905, ncgl0877, ncgl0905, ncgl0916, 115 ncgl1196, ncgl1202, ncgl1203, ncgl1208, ncgl1261, ncgl1262, ncgl1267, ncgl1301, ncgl1320, ncgl1322, ncgl1364, ncgl1366, ncgl1371,...

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Abstract

Provided are native promoters comprising polynucleotides isolated from Corynebacterium glutamicum, and mutant promoters derived therefrom, which may be used to regulate, i.e., either increase or decrease, on-pathway and / or off-pathway gene expression. Also provided are promoter ladders comprising a plurality of the promoters having incrementally increasing promoter activity. Also provided are hostcells and recombinant vectors comprising the promoters, and methods of expressing ancillary genes of interest and producing biomolecules using the host cells.

Description

[0001] Cross references to related applications [0002] This application requires the U.S. Provisional Application Serial No. 62 / 516,609 filed on June 7, 2017 under Article 119(e) and titled "Promoter from Corynebacterium glutamicum and Its Use in Regulating Auxiliary Gene Expression" The priority of, the disclosure of which is incorporated herein by reference in its entirety and used for the stated purpose. [0003] Sequence listing is incorporated by reference [0004] This application contains a sequence listing, which has been electronically submitted in ASCII format and incorporated herein by reference in its entirety. The ASCII copy was created on June 5, 2018 with the name ZMG-004_PCT_SL.txt and the size is 645,695 bytes. Technical field [0005] The present disclosure relates to natural promoters including polynucleotides isolated from Corynebacterium glutamicum and mutant promoters derived therefrom, host cells and recombinant vectors including the promoters, and modified ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/77
CPCC12N15/77C12P13/08C12N2310/3519C12N2330/51C12N2830/00C12N2830/55
Inventor Z·塞尔贝尔K·G·吉拉S·P·曼彻斯特P·埃涅尔特A·希勒
Owner ZYMERGEN INC
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