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Method for preparing inositol by multi-enzyme reaction system expressed by edible microorganisms

A multi-enzyme system and expression method technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as complex purification processes, and achieve the effect of reducing production costs

Active Publication Date: 2019-06-21
CHENGDU BOHAODA BIOLOGICAL TECH CO LTD
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Escherichia coli has the advantages of simple gene structure, easy genetic manipulation, and fast growth, but the products expressed by Escherichia coli may contain toxic proteins or antigenic proteins, as well as endotoxins produced by its cell wall, so the production of Escherichia coli of products require strict and complex purification processes

Method used

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  • Method for preparing inositol by multi-enzyme reaction system expressed by edible microorganisms
  • Method for preparing inositol by multi-enzyme reaction system expressed by edible microorganisms
  • Method for preparing inositol by multi-enzyme reaction system expressed by edible microorganisms

Examples

Experimental program
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Effect test

Embodiment 1

[0034] Embodiment 1: Lactic acid bacteria prepare multienzyme

[0035] The five genes whose KEGG numbers are ST0928, TM1168, TM0769, AF1794 and TM1415 were obtained from the official website of ATCC (www.atcc.org). Different primers were used to obtain from the corresponding genomic DNA by PCR, through Simple Cloning (You,C.,et al.(2012). "Simple Cloning via DirectTransformation of PCR Product (DNA Multimer) to Escherichia coli and Bacillus subtilis." Appl.Environ.Microbiol.78(5):1593-1595.) cloned into the pNZ9530 vector to obtain the corresponding gene expression vector, and then respectively transformed into lactic acid bacteria NZ9000 for protein expression. After detection, the target enzyme live negative.

Embodiment 2

[0036] Embodiment 2: Saccharomyces cerevisiae prepares multienzyme

[0037] The five genes were cloned into the pYES2 vector by the same method as in Example 1 to obtain the corresponding gene expression vectors, and then transformed into Saccharomyces cerevisiae Y187 for protein expression. The target enzyme activity was negative after detection.

Embodiment 3

[0038] Embodiment 3: Pichia pastoris prepares multienzyme

[0039] Using the same method as in Example 1, the five genes were cloned into the pPIC9K vector to obtain the corresponding gene expression vectors, and then transformed into Pichia pastoris GS115 for protein expression. The target enzyme activity was tested negative.

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Abstract

The invention discloses a method for preparing inositol by a multi-enzyme reaction system expressed by edible microorganisms. The method has the advantages that strains for the food industry serve asmulti-enzyme expression systems to produce isoamylase, glucan phosphorylase, glucose phosphate mutase, inositol-3-phosphate synthase and inositol monophosphatase, which are required for catalyzing starch and derivatives thereof, so that the possibility of contaminating the inositol by toxic protein, antigenic protein or endotoxin generated by escherichia coli in a production process is avoided fundamentally, a strict and complicated purification process is avoided, and the production cost is reduced.

Description

technical field [0001] The invention belongs to the field of enzyme-catalyzed production of inositol, and in particular relates to a method for preparing inositol by a multi-enzyme reaction system expressed by edible microorganisms. Background technique [0002] Inositol, also known as cyclohexanol, is one of the water-soluble vitamin B family, and its structure is shown in Formula I below. Inositol is an essential substance for the growth of humans, animals and microorganisms, and is widely used in medicine, food, feed and other industries. At present, the global demand is about 5,000 tons per year. Due to the current high price of inositol, the market prospect of inositol has not been fully developed. For example, the global feed production in 2013 was 9.6 billion tons. If 0.2- 0.5% inositol, then the output of inositol required by the feed industry should reach 1.9-4.8 million tons. In this case, the current domestic and even global production is far from meeting demand...

Claims

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Application Information

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IPC IPC(8): C12N15/75C12N15/77C12N9/44C12N9/10C12N9/90C12N9/16C12P7/18C12R1/125C12R1/15
Inventor 徐大勇黄彦菱蒲小平姜均王铎学
Owner CHENGDU BOHAODA BIOLOGICAL TECH CO LTD
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