Gellan gum lyase-producing bacteria and uses thereof

A technology of gellan gum and lyase, which is applied in the field of Pseudomonas, can solve the problems of slow cleavage rate and uncontrollable cleavage products, and achieve the effect of less environmental pollution

Active Publication Date: 2021-03-26
ZHEJIANG SCI-TECH UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, due to the high viscosity and high molecular weight of gellan gum, the further expansion of its application range is limited. The currently available industrial cracking methods for gellan gum are irradiation cracking and acid cracking, which reduce the gellan gum by corresponding physical or chemical means. Molecular weight, but both methods have defects such as slow cracking rate and uncontrollable cracking products, which limit the further development of its industrial application

Method used

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  • Gellan gum lyase-producing bacteria and uses thereof
  • Gellan gum lyase-producing bacteria and uses thereof
  • Gellan gum lyase-producing bacteria and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Embodiment 1: gellan gum lyase producing bacteria PE5 (Pseudomonas PE5)

[0017] The screening method is: configure the improved MM basic screening medium, use gellan gum as the only carbon source and coagulant, dissolve 1g of soil sample in 9ml 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 this screening have the ability to crack and use gellan gum, and the most obvious cavity in the plate is the gellan gum lyase producing bacteria PE5 (Pseudomonas PE5).

[0018] The strain PE5 was preserved under the preservation name of Pseudomonas hunanensis, preservation unit: General Microbiology Center of China Committee for the Collection of Microbial Strains, preservation address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, preservation number: CGMCC NO.16440 , the preservation time was September 07, 2018.

[0019] ...

Embodiment 2

[0030] Embodiment 2, the growth characteristics of bacterial strain PE5:

[0031] 1. Culture medium:

[0032] LB medium (LB liquid medium): 5 g of yeast extract, 10 g of peptone, 10 g of sodium chloride, 1000 ml of tap water, pH 7.4.

[0033] Improved MM basal medium: gellan gum 0.5%, ammonium nitrate 1.0g / L, potassium dihydrogen phosphate 0.5g / L, disodium hydrogen phosphate 1.5g / L, sodium chloride 1.0g / L, magnesium sulfate heptahydrate 0.2 g / L (put at the end), adjust the pH to 7.2.

[0034] That is, the improved MM basal medium is: 5g gellan gum, 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 add seven Magnesium sulfate water 0.2g.

[0035] Composition of PY fermentation medium: 0.2% gellan gum, 0.1% yeast extract, 0.2% peptone; pH7.0.

[0036] That is, the PY fermentation medium is: 2 g of gellan gum, 1 g of yeast extract, 2 g of peptone, 1000 ml of tap w...

Embodiment 3

[0045] Embodiment 3: utilize pseudomonas PE5 to produce the method for gellan gum lyase, carry out following steps successively:

[0046] 1) Transfer an inoculation loop of Pseudomonas PE5 to 100ml LB 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, and the supernatant (located on the upper layer) was collected.

[0049] 3) Add all the supernatant to a 10KDa pore size ultrafiltration tube at 7000rpm for 20min, use a pipette to absorb 5ml of Tris-HCl buffer solution pre-cooled at 4°C, 0.01M, pH7.0 to wash the ultrafiltration membrane, back and forth Aspirate se...

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Abstract

The present invention relates to a kind of Pseudomonas and its application. Specifically, the present invention discloses a gellan gum lyase producing bacterium PE5 (Pseudomonas PE5), which is Pseudomonas hunanensis, and its preservation number is: CGMCC NO :16440. The present invention also simultaneously discloses the use of the gellan gum lyase producing bacteria PE5: producing gellan gum lyase.

Description

technical field [0001] The invention relates to a pseudomonas (gellan gum lyase-producing bacteria) and its application. Background technique [0002] As a fermentation industrial product, microbial polysaccharides not only have excellent performance but also have many special functions that other similar polysaccharides lack. They are more widely used than animal and plant polysaccharides, and their production is not affected by factors such as geographical environment, climate, and natural disasters, and the production cycle is short. , the output and quality are stable, and the cost performance is high; to a large extent, it can meet people's demand for natural and pollution-free food. Gellan gum is a new type of microbial polysaccharide, which was discovered in 1978. It is a linear anionic heteropolysaccharide produced by the fermentation of Sphingomonas elodea under aerobic conditions. It has been approved as a food additive to be used in various foods in Japan, the Un...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/88C12R1/38
CPCC12N9/88C12N1/205C12R2001/38
Inventor 李昂李欧胡秀芳黄敬瑜胡婷婷骆航琦孟冯斌孟圆
Owner ZHEJIANG SCI-TECH UNIV
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