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

A kind of Bacillus xylosinolysine and its method for preparing α-keto acid

A technology of Bacillus glycolysine and Bacillus lysine, which is applied in the field of Bacillus xylosyllysine to prepare α-keto acid, which can solve the problem that the production intensity of α-keto acid is not high and it is difficult to realize industrialization problems such as broad substrate and product tolerance, broad substrate specificity, and less by-products

Active Publication Date: 2018-10-12
CHONGQING UNIV OF POSTS & TELECOMM
View PDF10 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although L-amino acid deaminase can specifically catalyze the deamination of L-amino acids without the generation of hydrogen peroxide, which avoids the further decarboxylation of α-ketoacids catalyzed by hydrogen peroxide, but the high concentration of L-amino acids and the product α-ketoacids are harmful to L-amino acids. - Significant inhibitory effect on amino acid deaminase, α-keto acid production is not high intensity, difficult to achieve industrialization

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 kind of Bacillus xylosinolysine and its method for preparing α-keto acid
  • A kind of Bacillus xylosinolysine and its method for preparing α-keto acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1: strain screening

[0028] (1) 20 soil samples collected from different areas were naturally air-dried and crushed, and 5 g of crushed soil samples were added to conical flasks containing 50 ml of sterile water, shaken (200 rpm) at 30 ° C for 20 min, and statically Take 1ml of the supernatant for gradient dilution, and then spread 50μl of the diluted solution on a petri dish loaded with solid medium, and incubate at 30°C for 3 days. The solid medium used for enrichment is: peptone 10g / L, beef extract 3g / L, sodium chloride 5g / L, agar 15g / L, pH7.2.

[0029] (2) Single colonies grown on the solid medium in step (1) were picked respectively, inoculated into 5 ml sterile liquid medium, and cultured on a shaker (200 rpm) at 30° C. for 24 hours. Take 1ml of the bacterial liquid, and use the 2,4-dinitrophenylhydrazine method to detect pyruvate. The positive strain was inoculated onto the solid medium in step (1), cultured at 30°C for 24h, then inoculated into 5ml...

Embodiment 2

[0030] Embodiment 2: bacterial strain identification

[0031] The strain with the highest L-alanine oxidative deamination activity was identified by 16s rDNA. The strain has 100% homology with Lysinibacillus xylanilyticus, and it was named Lysinibacillus xylanilyticus XX-2. See the appendix for the 16s rDNA sequence.

Embodiment 3

[0032] Embodiment 3: Bacterial strain culture and thalline harvest

[0033] The slant medium is: 3g / L beef extract, 5g / L yeast extract, 5g / L sodium chloride, 15g / L agar, pH6.0.

[0034] The seed medium is: 3g / L beef extract, 5g / L yeast extract, 5g / L sodium chloride, pH6.0.

[0035] Enzyme production medium: 3g / L beef extract, 5g / L yeast extract, 5g / L sodium chloride, 3g / L L-alanine, pH6.0.

[0036] Slant culture: Inoculate the screened Bacillus xylosinus XX-2 on the slant medium, and culture at a constant temperature of 30°C for 48 hours;

[0037] Seed culture: inoculate the strains cultivated on the slant with an inoculation loop in 5ml of fermentation medium under aseptic conditions, and culture on a shaker (180rpm) at 30°C for 48 hours to obtain a seed solution;

[0038] Shake flask culture: the seed solution was introduced into fresh fermentation medium with 10% inoculum, and cultured on a shaker (180 rpm) at 30° C. for 48 hours.

[0039] Bacterial cell harvesting: the ...

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 belongs to the technical field of biology and particularly relates to lysinibacillus xylanilyticus and a method for preparing alpha-ketoacid with the same. The lysinibacillus xylanilyticus XX-2 with a preservation number being CCTCC No:M2015520 is preserved in the China Center for Type Culture Collection. Whole cells of the lysinibacillus xylanilyticus XX-2 are taken as a biocatalyst to realize oxidative deamination of L-amino acids to obtain corresponding alpha-ketoacid, ammonia gas and hydrogen peroxide in the presence of air by L-amino acid oxidase in the lysinibacillus xylanilyticus XX-2, and in-situ decomposition of the generated hydrogen peroxide is realized by co-expressed catalase in the cells. The method overcomes defects of chemical synthesis methods and fermentation methods for preparing the alpha-ketoacid, has advantages of mild reaction conditions, environment friendliness, low cost, wide product spectrum and the like and is applicable to industrial production of the alpha-ketoacid.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a xylosyllysine bacillus and a method for preparing α-ketoacid by transforming L-amino acid with the strain. Background technique [0002] In organisms, α-keto acids are not only intermediates in sugar metabolism, but also participate in the biosynthesis of essential amino acids. α-keto acids are a class of bifunctional compounds, and are key intermediates in organic synthesis, drug synthesis and biosynthesis, and are widely used in biomedicine, food, cosmetics, feed and other fields. For example, four α-ketoacid calcium salts (α-ketoisocaproate calcium, α-ketoisovalerate calcium, α-keto-β-methylvalerate calcium, phenylpyruvate calcium), 1 hydroxyacid calcium salt (racemic hydroxymethionine calcium), a compound of 5 kinds of L-amino acids (L-lysine acetate, L-threonine, L-tryptophan, L-histidine, L-tyrosine) α-ketoacid tablets (Kaitong) are used to prevent and treat dam...

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): C12N1/20C12P7/40C12P7/50C12R1/01
Inventor 夏仕文熊文娟韦燕婵何从林徐红梅
Owner CHONGQING UNIV OF POSTS & TELECOMM
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