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Microbial production of alkanolamides and amidoamines and uses thereof

一种微生物体、脂肪酰胺的技术，应用在微生物、基于微生物的方法、散装化学品生产等方向，能够解决效率低下、经济和环境昂贵等问题

Pending Publication Date: 2021-03-19
GENOMATICA INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0014] These methods, and others known in the art for the synthesis of fatty amides, often involve inefficient reaction steps and are therefore expensive from both an economical and an environmental point of view

Method used

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  • Microbial production of alkanolamides and amidoamines and uses thereof
  • Microbial production of alkanolamides and amidoamines and uses thereof
  • Microbial production of alkanolamides and amidoamines and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0169]This example illustrates the construction of genetically engineered microorganisms in which the expression of acyl-CoA dehydrogenase, outer membrane protein receptor, pyruvate formate lyase, lactate dehydrogenase and transcriptional repressors is attenuated.

[0170] E. coli MG1655 DV4 is a genetically engineered E. coli K strain that contains fadE (acyl-CoA dehydrogenase), fhuA (outer membrane protein receptor), pflB (pyruvate formate lyase), and ldhA (lactate dehydrogenase enzyme) gene (see US Patent Application Publications 2011 / 0072714 and 2011 / 0162259, which are incorporated herein by reference). The fabR gene (GenBank accession number AAC76945) of Escherichia coli MG1655 encodes a transcriptional repressor, which was extracted from Escherichia coli MG1655 by the Lambda Red system according to Datsenko et al. (2000) Proc. DV4 deletions, modifications of the system are described herein.

[0171] The two primers used to generate the deletion strain were:

[0172] fa...

Embodiment 2

[0184] This example illustrates the construction of a genetically engineered microorganism in which the nucleotide sequences encoding thioesterase ('tesA) and acyl-CoA synthase (fadD) are integrated into the chromosome of the microorganism, and Under the control of an inducible promoter.

[0185] 'tesA is a nucleotide sequence comprising the E. coli tesA gene without a leader sequence (GenBank index number AAC73596, accession number U00096.2). 'tesA encodes Escherichia coli thioesterase (EC 3.1.1.5, 3.1.2.-), in which the first twenty-five amino acids are deleted, and the amino acid at position 26 - alanine is replaced by methionine replace. Thus, the methionine becomes the first amino acid of 'tesA (Cho et al. (1995) J. Biol. Chem. 270:4216-4219). E. coli fadD (GenBank Accession No. AAC74875; Accession No. U00096.2) encodes an acyl-CoA synthase.

[0186] Construction of pACYC-Ptrc plasmids containing 'tesA or 'tesA-fadD

[0187] The 'tesA gene was obtained from the pETDue...

Embodiment 3

[0210] This example illustrates the production of N-palmitoylethanolamine by expressing the gene encoding palmitoyl putrescine synthase in a genetically engineered microorganism.

[0211] The gene encoding bacterial palmitoyl putrescine synthase (PPS) was synthesized by DNA2.0 (Menlo Park, CA) and identified as GenBank accession number AY632377 (Brady, S.F., et al. (2004) J.Nat.Prod. , 67:1283-1286) (SEQ ID NO:2). The DNA2.0 plasmid - designated pJ201:30127 - was designed to contain NcoI sites adjacent to the start codon and PmeI sites adjacent to the stop codon. The PPS-encoding gene in pJ201:30127 was not codon-optimized. The PPS-encoding gene was subcloned into the expression vector OP80, which was described previously (see US Patent Application Publication 2010 / 0154293, which is hereby incorporated by reference in its entirety). The OP80 vector is based on pCL1920, a low-copy plasmid that expresses operably linked genes under the control of the IPTG-inducible trc promote...

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Abstract

The disclosure relates to a recombinant microorganism engineered to express an enzyme which catalyzes the conversion of a primary amine and an acyl thioester to a fatty amide. The disclosure further encompasses a method of producing a fatty amide by culturing the recombinant microorganism in the presence of a carbon source.

Description

[0001] This application is a divisional application of Chinese Invention Patent Application No. 201380026545.3 with the filing date of March 12, 2013, and the title of the invention is "Microbiological Production of Alkanolamides and Amides and Their Applications". [0002] Cross References to Related Applications [0003] This application claims the benefit of US Provisional Application No. 61 / 623,711 (filed April 13, 2012), the entire disclosure of which is incorporated herein by reference. [0004] sequence listing [0005] This application contains a Sequence Listing, which has been filed in ASCII via EFS-Web and is hereby incorporated by reference in its entirety. The ASCII copy was generated on March 11, 2013, named LS00041PCT_SL.txt and 69,769 bytes in size. [0006] field of invention [0007] The present disclosure relates to microbial organisms that have been genetically engineered to express enzymes that produce fatty amides when cultured in the presence of a carbo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N1/19C12N1/15C12N1/13C12P13/02C12R1/865C12R1/73C12R1/19C12R1/06C12R1/645C12R1/01C12R1/89C12R1/63C12R1/265C12R1/125C12R1/10C12R1/40C12R1/39C12R1/465
CPCC12N9/1029C12N9/18C12P13/02C12Y203/01086C12Y301/01005C12N9/16C12Y301/02014C12N15/63C12N15/70C12N15/74C12N15/79Y02P20/52
Inventor 贾森·J·鲁特斯斯蒂芬·德尔卡戴雷
Owner GENOMATICA INC
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