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Curdlan producing strain and application thereof

A technology of bacterial strain and natural rubber, applied in the application, bacteria, microorganism and other directions, can solve the problems of limited wide application, high production cost and poor rubber production performance.

Inactive Publication Date: 2018-12-18
EAST CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In my country, Shandong Zhongke Biotechnology Co., Ltd. has also realized the industrial production of curdlan gum, but the performance of curdlan gum produced is far behind that of Japan's Takeda company.
Due to the low yield and poor rubber production performance of domestic strains used for curdlan production, the production cost is high and the application effect is poor, which greatly limits its wide application in the market.

Method used

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  • Curdlan producing strain and application thereof
  • Curdlan producing strain and application thereof
  • Curdlan producing strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] The screening of embodiment 1 bacterial strain HS-5

[0087] Using Alcaligenes faecalis var.myxogenes 10C3 (IFO13714) as the starting strain, through ultraviolet mutagenesis, nitrosoguanidine mutagenesis, sodium nitrite mutagenesis and ARTP mutagenesis, the strain HS was obtained by plate primary screening and shake flask re-screening -5. The conditions of ultraviolet mutagenesis are: action time 45s, mutagenesis distance 30cm, dark treatment 1h. The conditions for mutagenesis of nitrosoguanidine are: concentration 1mg / ml, action time 30min, action pH 8.5. NIT mutagenesis conditions are: 1 mg / ml, action time 30 min, action pH 5.5. The ARTP mutagenesis conditions were as follows: the action time was 30s, the action power was 100W, and the action distance was 2mm.

[0088] Genetic stability verification: The high-yield strain HS-5 was continuously passaged on the slant medium for 10 times, each passage was respectively inserted into the fermentation medium, 200r / min, 3...

Embodiment 2

[0089] Identification of embodiment 2 bacterial strain HS-5

[0090] Physiological and biochemical identification: The screened high-yielding strain HS-5 was subjected to physiological and biochemical identification. The strain was Gram-negative bacteria, rod-shaped, and the colony shape on LB solid medium was round, smooth, slightly raised, and neatly edged. . Other identification results are listed in Table 1.

[0091] 16S rDNA gene amplification: Extract the strain HS-5 genome and perform 16S rDNA gene amplification. The primers for PCR amplification of 16S rDNA gene were 27f and 1492r, and their gene sequences were 27f, 5′-AGAGTTTGATCCTGGCTCAG-3′, 1492r, 5′-CGGTTACCTTGTTACGACTT-3′, respectively. The PCR amplification system is DNA template 2ul, Premix Ex Taq enzyme 25ul, Forward Primer (27f) 2ul, Downstream Primer (1492r) 2ul, ddH 2 O 19ul. The program of PCR amplification was pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30 s, annealing at 55°C for 30 s...

Embodiment 3

[0093] Fermentation and extraction of curdlan produced by embodiment 3 Agrobacterium.sp strain HS-5

[0094] Scrape an appropriate amount of HS-5 cells from the slant culture medium with an inoculation loop and insert it into the seed culture medium, place it in a shaker, 30°C, 200r / min, and cultivate for 18 hours to obtain a seed liquid, with an inoculum size of 8%. Insert it into the fermentation medium, place it in a shaker, 30° C., 200 r / min, and cultivate for 96 hours to obtain a fermentation broth.

[0095] The seed culture medium component is: glucose 20g / L, diammonium hydrogen phosphate 3g / L, potassium dihydrogen phosphate 1.5g / L, magnesium sulfate heptahydrate 1g / L, corn steep liquor 1g / L, calcium carbonate 2g / L, cultivate Base pH = 7.0.

[0096] The components of the fermentation medium are: glucose 70g / L, diammonium hydrogen phosphate 1.8g / L, potassium dihydrogen phosphate 1.5g / L, magnesium sulfate heptahydrate 1g / L, corn steep liquor 1g / L, calcium carbonate 2g / L, ...

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Abstract

The invention discloses a curdlan producing strain and an application thereof. The curdlan polysaccharide producing strain HS-5 is Agrobacterium sp., and a preservation number is CGMCC NO. 15477. TheAgrobacterium sp. strain HS-5 is subjected to fermentation and extraction to obtain the curdlan having excellent properties such as high yield, small molecular weight and good gel brittleness, and theindustrial production and market demand of the curdlan can be better satisfied.

Description

technical field [0001] The invention belongs to the technical field of microbial breeding, and in particular relates to a curdlan producing bacterial strain and application thereof. Background technique [0002] Curdlan gum is a new type of biopolymer polymer produced by Rhizobium sp. or Agrobacterium sp., which is composed of glucose structural units linked by β-1,3-D-glucosidic bonds. It is the third exopolysaccharide produced by microbial fermentation after xanthan gum and gellan gum approved by the US FDA for use in the food industry. It has unique thermal gelation, frost resistance, and dehydration resistance. And other excellent properties are used in food as stabilizer, coagulant and thickener, etc., which can obviously improve the quality of foods including soybean products, flour products, meat products, etc., and are deeply loved by the public. As early as 1968, Takeda Corporation of Japan conducted research on the production of curdlan gum and its application in ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P19/04A23L29/269A23L29/30C12R1/01
CPCA23V2002/00A23L29/271A23L29/30C12P19/04C12N1/205C12R2001/01A23V2200/202A23V2200/242A23V2200/23A23V2250/504
Inventor 金明飞陈璐高红亮常忠义
Owner EAST CHINA NORMAL UNIVERSITY
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