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(Sclerotium rolfsii)WSH-G01 and method for producing scleroglucan by fermenting (Sclerotium rolfsii)WSH-G01

A technology of Sclerotinia and glucan, applied in the field of microorganisms, can solve the problems of high production cost, waste of resources, low substrate utilization rate, etc., and achieve the effect of increasing substrate utilization rate and reducing cost significantly

Active Publication Date: 2018-08-24
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the carbon source for scleroglucan production is mainly sucrose. Although the yield reaches 26g / L, the substrate utilization rate is low, the production cost is high, and resources are seriously wasted.
The use of cornstarch and corn yellow pulp to produce scleroglucan is not suitable for all regions, and the subsequent processing of the product is cumbersome

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] The isolation and identification of embodiment 1 bacterial strain

[0024] The strains isolated earlier were inoculated on PDA medium, the genomic DNA of the strains was extracted, and the gene sequence of 18S rDNA was amplified (the universal primers for PCR amplification were NS1F and NS8R). The PCR product was identified by 1% agarose gel electrophoresis, and the result was good. The PCR amplification product was sent to Sangon Bioengineering (Shanghai) Co., Ltd. for sequencing, and the sequencing results were compared with BLAST sequences on the NCBI website, and it was determined to be Sclerotium rolfsii.

[0025] The key substantive difference between the Sclerotinia provided by the present invention and the existing Sclerotinia: Compared with the existing Sclerotinia, the Sclerotinia provided by the present invention is easier to use glucose to produce scleroglucan, and it is easier to produce scleroglucan when it contains the same concentration of sucrose In th...

Embodiment 2

[0026] The cultivation of embodiment 2 Sclerotin and the fermentation of scleroglucan

[0027] Fermentation medium (g / L): KH 2 PO 4 1.0; KCL 0.5; citric acid 1.5; MgSO 4 ·7H 2 O 0.5g / L; pH 4.0; add different amounts of glucose (45, 50, 55), NaNO 3 (2, 2.25, 2.5), yeast extract powder (0.2, 0.37, 0.5).

[0028] Shake flask culture: inoculate an appropriate amount of mycelium into a 250ml shake flask containing 50ml of seed medium, culture at 28°C and 220r / min for 72h, and transfer 5% of the inoculum to a 25ml glucose fermentation medium. In a 250ml shake flask, culture at 30°C and 220r / min for 96h.

[0029] Adjust the part components (as shown in Table 1) of fermentation medium respectively, all the other components remain unchanged:

[0030] Table 1 Composition adjustment and corresponding yield of fermentation medium

[0031] Numbering

[0032] The results show that the culture medium formula (g / L): glucose 55; NaNO 3 2.25; yeast extract powder 0.5; KH 2 P...

Embodiment 3

[0033] The impact of embodiment 3 different carbon source concentrations on the production of scleroglucan

[0034]The specific embodiment is the same as Example 2, the difference is that the fermentation medium formula is: sodium nitrate 2.2g / L, yeast extract 0.5g / L, KH 2 PO 4 1.0g / L, MgSO 4 ·7H 2 O 0.5g / L, citric acid 1.5g / L, KCl 0.5g / L, pH 4.5; different glucose additions (55, 75, 95g / L) were used to explore the effect of carbon source on the production of scleroglucan.

[0035] Batch culture in a 3L fermenter: Transfer the above seed liquid cultured at 28°C and 220r / min for 72h to a 500ml shake flask with 100ml of seed liquid at a 5% inoculum size, and transfer it to a 500ml shaker flask with 100ml of seed liquid at 28°C and 220r / min Cultivate under conditions for 48 hours, and then inoculate 5% inoculum in a 3L fermenter with 1.5L fermentation medium for 96 hours, set the temperature at 30°C, and stir at a speed of 350r / min.

[0036] Fermentation results such as fig...

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Abstract

The invention discloses (Sclerotium rolfsii)WSH-G01 and a method for producing scleroglucan by fermenting the (Sclerotium rolfsii)WSH-G01, and belongs to the technical field of microorganisms. The method has the advantages that the strain CCTCC NO:M2017646 can be fermented for 56h at the glucose carbon source concentration of 75 g / L, and realize the output of the scleroglucan of about 20 g / L, indicating that the output effect is obviously better than the output effect of the existing international method that the output of scleroglucan reaches 26 g / L after culturing for 72h under the culture condition of 150 g / L saccharose; the utilization rate of a substrate is increased by 58%, and the cost is obviously reduced; the production technology is better than the existing glucan production technology, and the industrialized application prospect is important.

Description

technical field [0001] The invention relates to a sclerotinum and a method for fermenting and producing scleroglucan, belonging to the technical field of microorganisms. Background technique [0002] Scleroglucan, also known as Sclerotin, is a high molecular weight, non-ionic branched glucan. It is a dextran with β-D-(1-3) glucopyranosyl as the main chain, and every three glucose is β-D-(1-6) glucopyranosyl as the side chain. Scleroglucan can be obtained by pure culture and fermentation of filamentous fungi of the genus Sclerotinia. [0003] Scleroglucan has high temperature resistance, various electrolytes (5% NaCl, 5% Na 2 SO 4 , 20% CaCl 2 etc.) and a wide range of properties such as pH. It is widely used in the food industry as a thickener, gel or stabilizer, and in the pharmaceutical industry as tablet coating, eye drops, etc. [0004] At present, microorganisms have been used to produce scleroglucan. However, because scleroglucan is usually fermented by fungi, th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P19/08A23L29/269A61K47/36C12R1/645
CPCA23V2002/00A61K47/36A23L29/269C12P19/08C12N1/145C12R2001/645A23V2200/242A23V2200/228A23V2250/5042
Inventor 周景文陈坚曾伟主高荣伟堵国成
Owner JIANGNAN UNIV
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