Highly active gellan gum oligosaccharide producing bacteria and its application

A technology of gellan gum and gum oligosaccharides, which is applied in the direction of bacteria, microorganisms, microorganisms, etc., can solve the problems of difficult control of oxidative degradation methods, difficult separation and purification of oligosaccharide products, and narrow industrial application prospects, etc. High antioxidant activity and strong cracking activity

Active Publication Date: 2021-03-30
ZHEJIANG SCI-TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In the application of gellan gum that has been developed, with the continuous increase of its application fields, the high molecular weight and high viscosity of gellan gum itself limit the further expansion of its application range, and the existing industrial cracking methods include chemical cracking And oxidative cracking, the polysaccharide degraded by chemical degradation method has a wide molecular weight distribution, and it is difficult to separate and purify oligosaccharide products with a certain molecular weight; the oxidative degradation method has the characteristics that the process is difficult to control
As a result, the prospects for industrial application are very narrow

Method used

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  • Highly active gellan gum oligosaccharide producing bacteria and its application
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  • Highly active gellan gum oligosaccharide producing bacteria and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Embodiment 1, Pseudomonas alternata PE2 (highly active gellan gum oligosaccharide producing bacteria PE2):

[0017] The screening method is as follows: configure the improved MM basic screening medium, use gellan gum as the only carbon source and coagulant, dissolve 1 g of seaweed sample in 9 ml of sterile water, spread it on the plate, and culture it in the incubator at 30°C. By decomposing gellan gum, obvious holes will be produced on the plate, and the formed strains produced by screening have the ability to crack and use gellan gum. Among them, the most obvious cavity in the plate is Pseudomonas alternata PE2 (highly active gellan gum oligosaccharides). Sugar producing bacteria PE2).

[0018] The strain PE2 has been preserved, and the preservation name is Pseudoalteromonas sp., preservation unit: General Microbiology Center of China Microbiological Culture Collection Management Committee, preservation address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District,...

Embodiment 2

[0030] Embodiment 2, the growth characteristics of bacterial strain PE2:

[0031] 1. Culture medium:

[0032] 2216E medium (2216E liquid medium): yeast extract 1g, peptone 5g, ferric phosphate 0.01g, sodium chloride 15g, tap water 1000ml, pH7.4.

[0033] Improved MM basal medium: gellan gum 0.5%, sea salt 2.5%, ammonium nitrate 1.0g / L, potassium dihydrogen phosphate 0.5g / L, disodium hydrogen phosphate 1.5g / L, sodium chloride 1.0g / L, seven Magnesium sulfate water 0.2g / L (put last), adjust the pH to 7.2.

[0034] That is, the improved MM basic medium is: 5g gellan gum, 25g sea salt, 1.0g ammonium nitrate, 0.5g potassium dihydrogen phosphate, 1.5g disodium hydrogen phosphate, 1.0g sodium chloride, 1000ml tap water, adjust the pH to 7.2; Finally, 0.2 g of magnesium sulfate heptahydrate was added.

[0035] Composition of PY fermentation medium: 0.2% gellan gum, 0.1% yeast extract, 0.2% peptone; 2.5% sea salt, pH 7.0.

[0036] That is, the PY fermentation medium is: 2 g of gella...

Embodiment 3

[0045] Embodiment 3, utilize Pseudomonas alternata PE2 to produce the method for gellan gum lyase, carry out the following steps successively:

[0046] 1) Transfer an inoculation loop of Pseudomonas alternata PE2 to 100ml 2216E liquid culture based on 200r / min, 30°C for enrichment culture, culture time is 24h; enrichment culture solution is obtained.

[0047] 2), the enriched culture solution obtained in step 1) was connected to the PY fermentation medium at 200r / min, 30°C for activation culture, the culture time was 36h, the enriched culture medium: PY fermentation medium = 1:9 Volume ratio;

[0048] The obtained activated culture solution was centrifuged at 8000 rpm for 15 minutes at 4° C. to obtain the supernatant liquid located in the upper layer and the bacterial cells located in the lower layer respectively.

[0049] 3) Take out the supernatant and keep the lower layer of bacteria, add 0.01M, pH7.0 Tris-HCl buffer 15ml to the bacteria to resuspend, centrifuge at 8000rpm...

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Abstract

The invention relates to a pseudomonas alternata (highly active gellan gum oligosaccharide-producing bacteria) and its application. Specifically, the present invention provides a Pseudomonas alteromonas PE2: Pseudoalteromonas sp. and its preservation number is: CGMCC NO: 16439. The present invention also simultaneously provides the application of the above-mentioned Pseudomonas alternata PE2: producing gellan gum lyase; producing gellan gum oligosaccharides.

Description

technical field [0001] The invention relates to a pseudomonas alternata (highly active gellan gum oligosaccharide-producing bacteria) and its application. Background technique [0002] Polysaccharide lyases are a class of degrading enzymes produced by prokaryotic or eukaryotic organisms that act on polysaccharides. The substrates of most polysaccharide lyases are negatively charged negative polysaccharides, which act on 1,4 glycosidic bonds linked to uronic acid through a β-elimination mechanism, and the final products are unsaturated oligosaccharides. In recent years, scholars at home and abroad have tried to use polysaccharide lyase to degrade large molecular weight polysaccharides into unsaturated oligosaccharides, thereby endowing oligosaccharides with new physiological activities and overcoming many defects of macromolecular polysaccharides. Gellan gum, as a typical negative polysaccharide, can also be degraded into unsaturated gellan gum oligosaccharides through the β...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P19/04C12P19/00C12R1/01
CPCC12P19/00C12P19/04C12N1/205C12R2001/01
Inventor 李欧李昂胡秀芳黄敬瑜高旭骆冠英黄筠骅宋诗雯刘雅琦
Owner ZHEJIANG SCI-TECH UNIV
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