Method for biologically preparing mannoglycerate

A technology of mannose and glyceric acid, which is applied in the field of biological preparation of mannose and glyceric acid, can solve problems such as low yield and failure to meet production requirements

Inactive Publication Date: 2020-10-20
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Related studies have used yeast strains as the starting strain to realize the fermentation method to synthesize mannose glyceric acid by constructing microbial recombinant strains, but the yield is very low, far from meeting the production requirements

Method used

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  • Method for biologically preparing mannoglycerate
  • Method for biologically preparing mannoglycerate

Examples

Experimental program
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Effect test

Embodiment 1

[0036] Example 1 In vitro multi-enzyme catalysis converts mannose into mannose glyceric acid

[0037] The catalytic pathway for the conversion of mannose to mannoglyceric acid by a multi-enzyme catalytic system in vitro see figure 1 . The key enzymes and key steps involved include: (1) Mannokinase, used to catalyze the phosphorylation of mannose to generate mannose 1-phosphate; (2) Mannose 1-phosphate glycosyltransferase, used to catalyze mannose- 1-phosphate and guanosine triphosphate (GTP) generate GDP-mannose and pyrophosphate; (3) mannose glycerate synthase, which is used to catalyze the synthesis of mannose glycerate from GDP-mannose and glyceric acid.

[0038]In this embodiment, the mannokinase is derived from Bifidobacterium longum, and the amino acid sequence is numbered B7GN78 (SEQ ID NO: 1) on UniProt; the mannose 1-phosphate glycosyltransferase is derived from Pyrococcus, and the amino acid sequence is in The number on UniProt is O58649 (SEQ ID NO: 2); the mannose...

Embodiment 2

[0042] Embodiment 2, construction Corynebacterium glutamicum recombinant strain MG1 and MG2

[0043] 1. The construction of the recombinant strain MG1 of Corynebacterium glutamicum comprises the following steps:

[0044] 1.1. Mannokinase MK (SEQ ID NO:1) derived from Bifidobacterium longum and mannose 1-phosphate glycosyltransferase MPG (SEQ ID NO:2) derived from Pyrococcus were introduced into Corynebacterium glutamicum ), mannose glycerate synthase MGS (SEQ ID NO: 3) derived from Halothermophilus marinum, to obtain a recombinant strain of Corynebacterium glutamicum, named strain MOS1.

[0045] The specific construction process is as follows:

[0046] Using the plasmids pUC57-MK, pUC57-MPG and pUC57-MGS as templates, the MK gene, MPG gene and MGS gene were obtained by PCR amplification, and connected to the vector pEC-XK99E by restriction enzyme ligation to obtain the recombinant plasmid pE -MK-MPG-MGS.

[0047] 1.2. Introduce the recombinant vector pE-MK-MPG-MGS into the ...

Embodiment 3

[0051] Embodiment 3, the application of Corynebacterium glutamicum recombinant strain MG1 and MG2 in the synthesis of mannose glyceric acid

[0052] The application of the recombinant strain of Corynebacterium glutamicum in the synthesis of mannose glyceric acid comprises the following steps:

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Abstract

The invention discloses a method for biologically preparing mannoglycerate, and relates to construction of an in-vitro multi-enzyme catalytic reaction system. The method comprises three enzymes, namely mannokinase, mannose-1-phosphate glycosyltransferase and mannoglycerate synthetase, and mannose and glycerate are converted to synthesize the mannoglycerate. The invention further discloses a construction method of recombinant corynebacterium glutamicum. The recombinant strain is applied to fermentation synthesis of the mannoglycerate, the mannoglycerate product is synthesized by taking the mannose and the glycerate as raw materials through a fermentation method, and the obtained mannoglycerate can be applied to the fields of foods, medicines, cosmetics and the like.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for biologically preparing mannose glyceric acid, which belongs to the field of biomanufacturing. Background technique [0002] Mannoglyceric acid is composed of a mannose molecule and a glyceric acid molecule connected. This compound is important for many high-temperature bacteria, maintaining its stable osmotic pressure under extreme conditions; at the same time, it also helps maintain the stability of protein structure , and the effect is better than trehalose, glycerol or any other known solvents. Recent experiments have shown that mannoglyceric acid can significantly enhance the immune system of the human body under in vitro conditions, and has the effect of treating diseases caused by protein misfolding or neurodegeneration (such as Alzheimer's disease and Parkinson's disease). At present, mannoglyceric acid is mainly extracted from seaweed, and the production cost of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/44C12N15/77C12N15/54C12N15/61C12R1/15
CPCC12P19/44C12N15/77C12N15/52C12N9/1205C12N9/1051C12N9/90C12Y207/01007C12Y204/01269C12Y504/02008
Inventor 杨建刚孙媛霞田朝玉曾艳马延和
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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