Unlock instant, AI-driven research and patent intelligence for your innovation.

A method for producing 2-oxo-α-d-glucopyranosyl ascorbic acid and its special engineering bacteria

A glucopyranosyl, ascorbic acid technology, applied in microorganism-based methods, biochemical equipment and methods, bacteria and other directions, can solve the problems of low conversion rate, poor specificity, increased enzyme cost, etc., and achieve mild reaction conditions and process. Simple route effect

Active Publication Date: 2021-08-24
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The main problems of using CGTase enzyme to produce AA-2G in the prior art are: (1) Usually α-cyclodextrin or β-cyclodextrin is used as the sugar donor, but the cost of α-cyclodextrin is too high, and β-cyclodextrin The solubility of refined alcohol is low; (2) poor specificity and low conversion rate for low-cost substrates such as maltodextrin and maltose; (3) the product obtained by using CGTase is AA-2Gn, which also requires the degradation of glucoamylase, which increases the enzyme activity. cost

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
  • A method for producing 2-oxo-α-d-glucopyranosyl ascorbic acid and its special engineering bacteria
  • A method for producing 2-oxo-α-d-glucopyranosyl ascorbic acid and its special engineering bacteria
  • A method for producing 2-oxo-α-d-glucopyranosyl ascorbic acid and its special engineering bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0104] Embodiment 1, construct recombinant bacteria and use recombinant bacteria to prepare AA-2G

[0105] 1. Construction of recombinant bacteria

[0106] 1. Extract the genomic DNA of Leuconostoc citrus.

[0107] 2. Using the genomic DNA obtained in step 1 as a template, perform PCR amplification with a primer pair composed of F1 and R1, and recover the PCR amplification product.

[0108] F1: 5'-GCCTGGTGCCGCGCGGCAGC CTCGAGATGGAAATTCAAAACAAAGCAATGC-3';

[0109] R1: 5'-CAGCTGCAGACCGAGCTCACC CTGCAG TTATTTGTTTTGTAAGACTGTCTTG-3'.

[0110] 3. Take the PCR amplification product obtained in step 2, perform double digestion with restriction endonucleases XhoI and PstI, and recover the digested product.

[0111] 4. Digest the vector pBAD / HisB with restriction endonucleases XhoI and PstI to recover a vector backbone of about 4000 bp.

[0112] 5. Ligate the digested product of step 3 with the vector backbone of step 4 to obtain recombinant plasmid A.

[0113] According to the s...

Embodiment 2

[0144] Example 2, constructing recombinant bacteria and using recombinant bacteria to prepare AA-2G

[0145] 1. Construction of recombinant bacteria

[0146] The recombinant plasmid C was prepared. According to the sequencing results, the structure of the recombinant plasmid C is described as follows: the small fragment between the XhoI and PstI restriction sites of the vector pBAD / HisB is replaced by the double-stranded DNA molecule shown in sequence 4 of the sequence table. The DNA molecule shown in sequence 4 of the sequence listing encodes the protein shown in sequence 3 of the sequence listing. The double-stranded DNA molecule shown in sequence 4 of the sequence listing is amplified from the genome DNA of Bifidobacterium adolescentis.

[0147] The recombinant plasmid C was introduced into Escherichia coli BW25113 to obtain the recombinant plasmid C.

[0148] 2. Application of recombinant bacteria to prepare AA-2G (one-step enzymatic method)

[0149] Recombinant bacter...

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 a method for producing 2-oxy-α-D-glucopyranosyl ascorbic acid and special engineering bacteria. The invention provides a method for producing 2-oxygen-α-D-glucopyranosyl ascorbic acid, comprising the following steps: using ascorbic acid substances and sucrose as raw materials, under the action of engineering bacteria, producing 2-oxygen-α ‑D‑glucopyranosyl ascorbic acid; the engineering bacterium is a recombinant bacterium expressing a functional protein, which is obtained by introducing the functional gene encoding the functional protein into the starting bacterium; the functional protein is shown in sequence 1 of the sequence table The protein or the protein shown in sequence 3 of the sequence listing; the ascorbic acid substance is ascorbic acid or ascorbate. The method provided by the invention uses cheap sucrose and ascorbic acid as raw materials to realize the one-step enzymatic production of AA-2G, and has the remarkable characteristics of mild reaction conditions, no pollution, simple process route, and suitable for large-scale industrial production.

Description

technical field [0001] The invention relates to a method for producing 2-oxo-α-D-glucopyranosyl ascorbic acid and special engineering bacteria. Background technique [0002] 2-Oxo-α-D-glucopyranosyl ascorbic acid (2-O-α-D-Glucopyranosyl-L-ascorbic acid, AA-2G) is an important biological compound. [0003] AA-2G is an important class of derivatives of ascorbic acid (vitamin C, L-ascorbic acid, AA). It is a compound formed by modifying the hydroxyl group on the C2 position of AA with a glucose group. , the oxidation reaction of AA is not easy to occur, so it is particularly stable in aqueous solution, and has no direct reduction, which effectively protects its biological activity. [0004] AA-2G has the same collagenolytic activity as AA, which can be used to enhance the synthesis of collagen in human skin cells, and can also prevent and treat acute diseases caused by ultraviolet radiation. Under the combined action of the enzyme system in the human body, AA-2G is easily dec...

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 Patents(China)
IPC IPC(8): C12P19/60C12N1/21C12R1/19
Inventor 胡美荣王雷彭颖陶勇
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI