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

Method for producing 5'-guanylic acid

A technology of guanylic acid and guanylic acid synthase, which is applied in the field of 5’-guanylic acid production and can solve problems such as difficulty in decomposition

Inactive Publication Date: 2011-03-30
AJINOMOTO CO INC
View PDF14 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still many unknowns about nucleotide decomposing enzymes, and several nucleotide decomposing enzymes have been found, and it is known that the yield can be improved by making them missing (Patent Documents 6 and 7), but complete inhibition of their degradation is difficult, it's a big problem

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
  • Method for producing 5'-guanylic acid
  • Method for producing 5'-guanylic acid
  • Method for producing 5'-guanylic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0120] Construction of nagD gene disruption strain derived from JM109 strain

[0121] Using the Escherichia coli JM109 strain used as a host for conventional DNA cloning as a parent strain, a NagD protein non-producing strain was first constructed. The NagD protein is encoded by the nagD gene (GenBank Accession No. X14135 SEQ ID NO: 11).

[0122] The deletion of nagD gene was first developed by Datsenko and Wanner called "Red-driven integration (Red-driven integration)" method (Proc.Natl.Acad.Sci.USA, 2000, vol.97, No.12, p6640- 6645) and λ phage-derived excision system (J.Bacteriol.2002Sep; 184(18):5200-3.Interactions between integrase and excisionase in the phage lambda excisive nucleoprotein complex. Cho EH, Gumport RI, GardnerJF.) to carry out . According to the "Red-driven integration" method, synthetic oligonucleotides in which a part of the gene of interest is designed on the 5' side and a part of the antibiotic resistance gene on the 3' side are used as primer...

Embodiment 2

[0155] Construction of ushA gene disrupted strains derived from JM109 strain and JM109ΔnagD strain

[0156] Using the JM109 strain and the JM109ΔnagD strain obtained in Example 1 as parent strains, a UshA non-producing strain was constructed. UshA is encoded by the ushA gene (GenBank Accession No. X03895SEQ ID NO: 14). According to the aforementioned method for disrupting the nagD gene, the ushA gene was disrupted using the primers of SEQ ID NO: 21 and 22 as primers for disrupting ushA. Thus, JM109ΔushA and JM109ΔushAΔnagD were obtained.

[0157] Introduction of GMP synthetase expression enhanced plasmid pSTV29-Ptac-guaA into JM109ΔushA strain and JM 109ΔushAΔnagD strain

[0158] The GMP synthetase expression enhancing plasmid pSTV29-Ptac-guaA described in Example 1 was introduced into the JM109ΔushA strain and the JM109ΔushAΔnagD strain to obtain the JM109ΔushA / pSTV29-Ptac-guaA strain and the JM109ΔushAΔnagD / pSTV29-Ptac-guaA strain respectively.

[0159] Trans...

Embodiment 3

[0163] Construction of aphA gene disrupted strains derived from JM109ΔushA strain and JM109ΔushAΔnagD strain

[0164] Using the JM109ΔushA strain and the JM109ΔushAΔnagD strain obtained in Example 2 as parent strains, an AphA non-producing strain was constructed. AphA is encoded by the aphA gene (GenBank Accession No. X86971 SEQ ID NO: 17). According to the aforementioned nagD gene disruption method, using the primers of SEQ ID NO: 23 and 24 as primers for aphA disruption, aphA gene disruption was carried out. Thus, JM109ΔushAΔaphA and JM109ΔushAΔaphAΔnagD were obtained.

[0165] Introduction of GMP synthetase expression enhanced plasmid pSTV29-Ptac-guaA into JM109ΔushAΔaphA strain and JM109ΔushAΔaphAΔnagD strain

[0166] The GMP synthetase expression enhancing plasmid pSTV29-Ptac-guaA described in Example 1 was introduced into the JM109ΔushAΔaphA strain and the JM109ΔushAΔaphAΔnagD strain to obtain the JM109ΔushAΔaphA / pSTV29-Ptac-guaA strain and the JM109ΔushAΔaph...

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 relates to a method for producing 5'-guanylic acid. 5'-guanylic acid (GMP) is produced efficiently by allowing a microorganism to react with xanthylic acid (XMP), wherein said microorganism is able to convert xanthylic acid into 5'-guanylic acid and has been modified so that the nagD gene does not function normally and 5'-guanylic acid synthetase activity is enhanced.

Description

technical field [0001] The present invention relates to a method for producing 5'-guanylic acid and novel microorganisms used in the production of 5'-guanylic acid. 5'-guanylic acid is useful as seasonings, medicines, and raw materials for seasonings and medicines. Background technique [0002] As an industrial preparation method of 5'-guanylic acid (guanosine-5'-monophosphate, hereinafter also referred to as "GMP"), guanosine is produced by fermentation method, and the obtained guanosine is phosphorylated by enzymatic method to obtain Methods of 5'-guanylic acid (Patent Documents 1 to 4). [0003] In addition, a production method is also known in which a microorganism belonging to the genus Escherichia having increased GMP synthetase activity is cultured with the following Brevibacterium ammoniagenes in a medium containing XMP and ammonia gas or glutamine , convert XMP into GMP with high efficiency, and generate and accumulate GMP in the culture solution, wherein the aden...

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): C12P19/32C12N1/21C12N15/52C12R1/01C12R1/07C12R1/15C12R1/185C12R1/19
CPCC12N9/0006C12P19/32C12R2001/07C12R2001/15C12R2001/19Y10S435/832Y10S435/843Y10S435/849
Inventor 深田宽朗浅原贵之桥口贤一
Owner AJINOMOTO CO INC
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