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

Construction method and application of homoserine and threonine biosynthetic pathway

A kind of homoserine, homoserine kinase technology, applied in industrial bio-economy and manufacturing, amino acid industrial production field

Pending Publication Date: 2022-03-01
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The inventors especially noticed that formaldehyde can react with pyruvate, the end product of glycolysis, in the non-natural pathway Homoserine cycle (Metab Eng, 2020. 60: 1-13) (ACSCatal, 2017. 7(3): 1707-1711) to generate 4-hydroxydiketone butyric acid, and the present invention believes that this provides a research idea for solving the problem of high energy consumption using carbon dioxide

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
  • Construction method and application of homoserine and threonine biosynthetic pathway
  • Construction method and application of homoserine and threonine biosynthetic pathway
  • Construction method and application of homoserine and threonine biosynthetic pathway

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] Example 1 - construction scheme of synthetic pathway

[0082] This new approach innovatively replaces low-energy CO with high-energy-density methanol / formaldehyde one-carbon compounds. 2 Introduced into the synthesis pathway of homoserine and threonine, it still uses glucose as the main carbon source for amino acid fermentation, and uses methanol as a one-carbon raw material to synthesize the intermediate metabolite 4-hydroxy- 2-ketobutyric acid, which then synthesizes homoserine through a transamination reaction, and homoserine can then synthesize threonine through natural pathways. see new route figure 1 . The unnatural nature of this new pathway is mainly reflected in the design and combination of different reactions in the first three steps to generate homoserine, while the latter two steps come from the natural threonine synthesis pathway to convert homoserine into threonine. Methanol and pyruvate can be converted into threonine through an in vitro multi-enzyme...

Embodiment 2

[0083] Example 2 Screening of Methanol Dehydrogenase

[0084] Methanol dehydrogenases from different sources were selected for enzyme kinetics characterization. It mainly includes seven NAD-dependent methanol dehydrogenases and two mutants that have been reported: derived from methylotrophic Bacillus methanolica ( Bacillus  methanolicus ) of methanol dehydrogenase: B m Mdh (SEQ ID NO.28), B m Mdh2 (SEQ ID NO.29), B m Mdh3 (SEQID NO.31) and B m Mdh2 mutants: B m Mdh2 mut (SEQ ID NO.30); derived from Bacillus stearothermophilus ( B.  stearothermophilus ) of alcohol dehydrogenase Bs Mdh (SEQ ID NO.32); derived from methylotrophic bacteria ( Cupriavidus  necator ) of alcohol dehydrogenase Cn Mdh2 (SEQ ID NO.33) and its mutant CT4-1 (SEQ ID NO.34); derived from Lysinobacillus xylan ( Lysinibacillus xylanilyticus ) of methanol dehydrogenase Lx Mdh (SEQ IDNO.35); Derived from Corynebacterium glutamicum ( Corynebacterium glutamicum R) methanol dehydrogena...

Embodiment 3

[0089] Example 3 - Screening of aldolase

[0090] Different types of aldolases were selected for enzyme kinetics characterization. Mainly including Escherichia coli ( Escherichia   coli ) of 2-keto-3-deoxy-L-rhamnolate aldolase Ec RhmA (type II aldolase), 2-dehydro-3-deoxy-D-gluconate aldolase Ec YagE (EC:4.1.2.51 Gene ID:944925) and dihydrodipicolinate synthase EcYjhH (SEQ ID NO. 37). The above three aldolases were constructed on the expression vector pET16b, because 2-keto-3-deoxy-L-rhamnolate aldolase Ec RhmA has the problem of inclusion bodies, so the MBP-RhmA method expressed as a fusion protein was designed: connect maltose binding protein (MBP) to the N-terminal of the RhmA sequence, and finally obtain the corresponding vector: pET16b- Ec RhmA MBP ,pET16b- Ec YagE, pET16b- Ec YjhH.

[0091] Then, the above-mentioned expression vectors were respectively transformed in Escherichia coli expression hosts E .coli  BL21(DE3), cultured at 37℃ to  OD 600 Whe...

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

The invention discloses construction of a new non-natural biosynthesis way of homoserine and threonine, and belongs to the technical field of industrial biology. The non-natural biosynthetic pathway comprises methanol dehydrogenase, aldolase, transaminase, homoserine kinase and threonine synthetase, and the pathway can be used for synthesizing homoserine or threonine by taking methanol and pyruvic acid as substrates. According to the present invention, the homoserine or the threonine can be synthesized by using methanol and pyruvic acid as the carbon source by constructing the in vitro multi-enzyme catalysis system and assembling in the microbial cell body through the pathway, and the pyruvic acid can be synthesized through glycolysis by using glucose as the substrate in the microbial cell body; therefore, homoserine or threonine is synthesized by taking methanol and glucose as carbon sources. According to the invention, the methanol-carbon compound with high energy density is introduced into a homoserine and threonine synthesis route, 200% of sugar-acid molar conversion rate can be theoretically realized, and the method has a great application prospect.

Description

technical field [0001] The present invention relates to the technical field of amino acid industrial production, in particular to the construction method and application of homoserine and threonine biosynthetic pathways, more specifically to the production of homoserine and threonine using methanol / formaldehyde and glucose / pyruvate as co-substrates The method belongs to the field of industrial bioeconomics and manufacturing. Background technique [0002] Homoserine and threonine are amino acids with important uses, mainly used in medicine, pesticide precursors, chemical reagents, food fortifiers, feed additives, etc. With the continuous increase of amino acid market demand, industry competition is also becoming increasingly fierce. The production of strains is the core of industrial fermentation, and major threonine manufacturers are actively improving the fermentation performance of strains and reducing production costs in order to maintain market competitiveness. How to ...

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
IPC IPC(8): C12P13/06C12P13/08C12P7/24C12P7/42C12N9/04C12N9/88C12N9/10C12N9/12C12N1/21C12N15/31C12R1/19
CPCC12P13/06C12P13/08C12P7/24C12P7/42C12N9/0006C12N9/88C12N9/1096C12N9/1205C07K14/245C12Y101/01006C12Y206/01001C12Y207/01039C12Y402/03001
Inventor 董红军李梁坡周海川谢永辉
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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