Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Production of muconic acid from genetically engineered microorganisms

A genetic modification and microbial technology, applied in biochemical equipment and methods, using vectors to introduce foreign genetic material, lyases, etc., can solve the problems of expensive, unusable, unstable multi-copy plasmids, etc.

Inactive Publication Date: 2014-12-17
MYRIANT CORP
View PDF27 Cites 9 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Multicopy plasmids are often too unstable to be used in large-scale industrial methods
Moreover, at least one of the genes on the plasmid starts from the promoter P tac expression, which requires isopropylthiogalactopyranoside (IPTG) or lactose to induce, and these two chemicals are too expensive to allow an economically attractive approach

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Production of muconic acid from genetically engineered microorganisms
  • Production of muconic acid from genetically engineered microorganisms
  • Production of muconic acid from genetically engineered microorganisms

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0097] Increased expression of aroG and aroF

[0098] The tyrR gene of E. coli can be mutated by any of several well-known methods, such as chemical or radiation mutagenesis and selection (e.g., by PCR and DNA sequencing) or selection for analogue resistance (e.g., to 4-fluorotyramide Acid resistance), transposon mutagenesis, bacteriophage Mu mutagenesis or transformation. In preferred embodiments, the mutation in the tyrR gene is a null mutation (a mutation that results in no detectable activity), while in a more preferred embodiment, at least part of the tyrR gene is deleted. This can be achieved, for example, by utilizing a two-step transformation method using linear DNA molecules (Jantama et al., 2008a; Jantama et al., 2008b). In the first step, cam R The , sacB cassette is integrated at the tyrR locus to replace most or all of the tyrR open reading frame. In a second step, a linear DNA including a deletion of the tyrR gene was integrated by double panel selection for r...

Embodiment 2

[0101] Feedback-resistant AroG and AroF

[0102]Mutations in the aroG gene of the AroG enzyme (3-deoxy-D-arabinoheptonate-7-phosphate synthase or DAHPS) leading to feedback resistance are known in the art (Shumilin et al., 1999; Kikuchi et al. , 1997; Shumilin et al., 2002). Also known are methods to generate, identify and characterize these mutations (Ger et al., 1994, Hu et al., 2003). Preferred mutations are those that confer complete resistance to phenylalanine inhibition. Introduction of any of the known published feedback resistance mutations into the aroG gene contained in a chromosome or on a plasmid can be done by any of several known methods, an example of which is mutagenesis PCR, wherein the desired Mutations were synthesized as part of PCR priming oligonucleotides (Hu et al., 2003). Correct placement of mutations was confirmed by DNA sequencing. The sequence of the wild type aroG gene from E. coli C is given in SEQ ID No.18. A preferred mutation is a point mu...

Embodiment 3

[0110] Deletion of aroE and muconate production from chromosomal DNA

[0111]In this example the effect of overexpressing aroB and aroG on a multi-copy plasmid and expressing genes encoding proteins functional in the muconate pathway was investigated. Strain MYR34, containing a deletion in the aroE gene encoding shikimate dehydrogenase, was used as the parental strain in these studies. Deletion of the chromosomal copy of aroE was accomplished in a manner similar to that described in Example 1 above. When MYR34 was transformed with the plasmid pCP32AMP overexpressing the aroG gene encoding the DAHP synthase protein functional in the shikimate pathway, DHS accumulation was significantly enhanced. When MYR34 was transformed with a plasmid expressing the aroB gene from a constitutive promoter, no significant increase in DHS accumulation was noted. However, when E. coli strain MYR34 was transformed with a plasmid expressing both aroB and aroG genes, there was increased accumulati...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

This present invention is in the field of producing renewable chemical feedstocks using biocatalysts that have been genetically engineered to increase their ability to convert renewable carbon resources into useful compounds. More specifically, the present invention provides a process for producing muconic acid form renewable carbon resources using a genetically modified organism.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of priority to US Provisional Application Serial No. 61 / 632,777, filed January 30, 2012. field of invention [0003] The present invention is in the field of the production of renewable chemical feedstocks using biocatalysts that have been genetically modified to increase their ability to convert renewable carbon sources into useful compounds. More specifically, the present invention provides methods for the production of muconic acid isomers from renewable carbon sources using genetically engineered biocatalysts. Background of the invention [0004] Adipic acid is a bulk chemical used in the manufacture of nylon 66. Adipic acid is currently produced from petrochemicals, but the synthesis is not environmentally friendly (Niu et al., 2002). Alternatively, adipic acid can be prepared by chemical hydrogenation of any of the three isomers (cis, cis; cis, trans; trans, trans) of muconic...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/63C12N15/70C12P7/40
CPCC07K14/245C12N9/0006C12N9/0016C12N9/0069C12N9/1022C12N9/1085C12N9/88C12N15/52C12P7/44C12Y101/01025C12Y205/01054C12Y402/01118C12N9/1205C12N1/20
Inventor R·R·约卡姆龚巍S·多勒R·西勒斯M·甘地J·G·佩罗
Owner MYRIANT CORP
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com