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Gluconacetobacter xylinus strain capable of producing free glucuronic acid

A technology of glucuronic acid and gluconacetobacter, applied in the direction of bacteria, microorganism-based methods, microorganisms, etc., to achieve the effects of easy popularization and application, high raw material utilization rate, and environmental protection

Active Publication Date: 2015-02-04
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, researchers at home and abroad have detected the presence of glucuronic acid in kombucha beverages in different parts of the world, but they are all fermented by mixed flora, and the use of a single microbial strain to ferment glucuronic acid is still mostly in the research stage

Method used

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  • Gluconacetobacter xylinus strain capable of producing free glucuronic acid
  • Gluconacetobacter xylinus strain capable of producing free glucuronic acid
  • Gluconacetobacter xylinus strain capable of producing free glucuronic acid

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Experimental program
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Effect test

Embodiment 1

[0028] The screening of embodiment 1 bacterial strain

[0029] (1) Strain screening

[0030] Take a small amount of kombucha fungus film to enrich in GY medium, culture at 30°C, 200r / min shaker for 48h, and then dilute to 10 -3 、10 -4 、10 -5 、10 -6 、10 -7 For 5 gradients, 200 μL of each diluted sample was applied to GYC solid medium containing 100 mg / mL natamycin, and each dilution gradient was repeated twice. Pick a single colony and incubate at 30°C for 72h. Select a single colony with a transparent circle and inoculate it on GY medium, cultivate it on a shaker at 30°C and 200 r / min until the logarithmic phase, collect samples and store it, collect the fermentation broth after 72 hours of cultivation, centrifuge, and take the supernatant for detection. A Q1 strain with higher yield was screened from many screened strains for further strain identification.

[0031] (2) Bacteria identification

[0032] The genomic DNA of the starting strain Q1 was extracted, and the 1...

Embodiment 2

[0036] Embodiment 2 Qualitative detection of glucuronic acid

[0037] After activation on the solid GY medium of Gluconacetobacter xylosus Q1, inoculate it into a 250mL Erlenmeyer flask containing 25mL of GY medium, culture it at 30°C and 200rmp / min for 72h, collect the fermentation broth, centrifuge, and take the The clear liquid was prepared for detection; the content of glucuronic acid in the fermentation liquid was detected by ion chromatography.

[0038]Determine the LC-MS spectrum of the glucuronic acid standard sample, and dilute the supernatant of the fermentation broth of G.xylinus Q1 by 10 times for LC-MS detection, and compare the two for spectrum analysis, as shown in figure 1 As shown, the molecular weight of the glucuronic acid standard sample was 193.0369 in the mass spectrometry scan at 2 minutes, and the sample detected substances with the same molecular weight (193.0370) at the same time, confirming that the fermentation broth contained glucuronic acid.

Embodiment 3

[0039] Determination of glucuronic acid detection method in embodiment 3 exopolysaccharide:

[0040] The bacteria were cultured in GY medium at 30°C and 200r / min for 72h, centrifuged at 8000rpm for 15min, and the supernatant was taken. Ethanol was added to the supernatant with 4 times the volume of the supernatant, and the mixture was kept at 4 °C overnight. The precipitate was collected by centrifugation at 8000r / min for 15 minutes. The precipitate was dried, and 0.5 mol / L H of 5 times the volume of the supernatant was added 2 SO 4 Solution, reacted at 70°C for 3h. The hydrolyzate was filtered through a 0.45 μm filter membrane, and the product was detected by LCMS. The detection result showed that the exopolysaccharide did not contain glucuronic acid.

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Abstract

The invention discloses a Gluconacetobacter xylinus strain capable of producing free glucuronic acid, belonging to the field of bioengineering. The screened Gluconacetobacter xylinus Q1 strain is collected by China Center for Type Culture Collection on July 23rd, 2014, and the collection number is CCTCC M2014353. The Gluconacetobacter xylinus strain is screened and separated from tea fungus, and can produce glucuronic acid by fermentation by using glucose as the substrate; and the free glucuronic acid content under simple fermentation conditions is 12.96 mg / L or so.

Description

technical field [0001] The invention relates to a strain of Gluconacetobacter xylosus producing free glucuronic acid, which belongs to the technical field of fermentation. Background technique [0002] Glucuronic acid, English name Glucuronic acid, is the uronic acid formed by the oxidation of the C-6 hydroxyl group of glucose into carboxyl. Glucuronic acid is a traditional liver detoxifier and immune function regulator. Reactive aldehyde groups and carboxyl groups can interact with groups in human endogenous and exogenous toxic substances such as hydroxyl, carboxyl, sulfhydryl, amino, etc., thereby increasing the water solubility of toxic substances and making them easy to excrete with urine and bile in vitro. [0003] Glucuronic acid can be used as an intermediate to synthesize many important medicines such as D-calcium glucarate and L-ascorbic acid. In addition, glucuronic acid is also commonly used as an additive in functional beverages, weight-loss drugs, cosmetics, e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P19/02C12R1/01
CPCC12P19/02C12N1/205C12R2001/01
Inventor 方芳刘晓慧仇钰莹陈坚
Owner JIANGNAN UNIV
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