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A strain of Sclerotinia sclerotiorum and its application in fermentative production of scleroglucan

A technology of neat Sclerotinia and scleroglucan, applied in the direction of fermentation, microbial-based methods, microorganisms, etc., can solve the problems of complex process control, insufficient stability, long fermentation time, etc., and achieve high viscosity, low price, Simple operation effect

Active Publication Date: 2017-05-03
SHANDONG FOOD & FERMENT IND RES & DESIGN INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the currently reported studies on the fermentation and production of scleroglucan focus on the optimization of the fermentation medium, but it still has the defects of long fermentation time (more than 96h), low yield (about 20g / l), and high production cost.
Although the published domestic and foreign patents CN201110006769.0, CN201310283916.8 and EP 1417325A2 on improving the fermentative production of scleroglucan have a maximum yield of 32g / L, the process control is complex and the stability is insufficient

Method used

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  • A strain of Sclerotinia sclerotiorum and its application in fermentative production of scleroglucan
  • A strain of Sclerotinia sclerotiorum and its application in fermentative production of scleroglucan
  • A strain of Sclerotinia sclerotiorum and its application in fermentative production of scleroglucan

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Embodiment 1: the selection and breeding of neat small sclerotium (Sclerotium rolfsii) SCL2010

[0056] Collect the sclerotium sclerotia from potato plants and tubers infected with white silk disease in Shouguang City, Gaomi City and Linqu County, Weifang City, Shandong Province, use the sclerotium as the separation material, and inoculate the sclerotia on the PDA solid medium under sterile conditions After culturing, after the sclerotia germinates and mycelium grows out, it is separated and purified, and then transferred to PDA slant medium for storage.

[0057] Transfer the hyphae separated and purified on the PDA medium to new PDA plates, culture at 28°C for continuous observation, and wait until the mycelium grows to cover the plate and form sclerotium until the sclerotia changes color and matures, record Mycelial growth, tuberculosis and sclerotia changes, combined with microscope observation to determine the Sclerotinia strain and number the strain; transfer the s...

Embodiment 2

[0062] Embodiment 2: Gene sequence sequencing of 18S rDNA of Sclerotium rolfsii (Sclerotium rolfsii) SCL2010 strain

[0063] The Sclerotium rolfsii SCL2010 strain obtained in Example 1, that is, the CCTCC NO.M2014325 strain, was entrusted to BGI for sequencing.

[0064] experimental method:

[0065] (1) Wash a certain amount of new sclerotia into the seed medium with physiological saline, culture at 28°C for 36 hours, after the sclerotia germinate and produce a large number of mycelia, centrifuge at 8000r / min to obtain mycelium;

[0066] (2) Using the Takara9765 Fungal DNA Extraction Kit to extract the genome from the obtained mycelia and send it to a sequencing company for sequencing.

[0067] Sequencing results: the length of the 18S rDNA gene sequence of Sclerotium rolfsii SCL2010 strain is 1694bp, and the specific nucleotide sequence is shown in SEQ ID NO.1.

[0068] By using the BLASTN program of the National Center for Biotechnology Information (NCBI), it was found tha...

Embodiment 3

[0070] Embodiment 3: the method for fermenting and producing scleroglucan of Sclerotium rolfsii (Sclerotium rolfsii) SCL2010 bacterial strain

[0071] (1) Inoculate the mature sclerotia of Sclerotinia sclerotiorum CCTCC No.M2014325 on the slanted solid medium of an eggplant-shaped bottle, and culture it statically at 30-34° C. for 5-7 days to obtain a large number of mature sclerotia for seed cultivation.

[0072] (2) Wash the mature sclerotia of Sclerotinia sclerotiorum CCTCC No.M2014325 with sterile physiological saline, inoculate an appropriate amount in the seed medium, and culture with aeration and stirring at 34° C. for 30 h.

[0073] (3) Scleroglucan fermentation production process:

[0074] Inoculate an appropriate amount of Sclerotinia sclerotinum CCTCC No.M2014325 matured through seed culture into the fermentation medium as a fermentation strain, and aerate and stir to control fermentation to produce scleroglucan.

[0075] Preferred fermentation control conditions: ...

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Abstract

The invention discloses a strain of Sclerotium rolfsii suitable for industrial production of scleroglucan. The strain is named Sclerotium rolfsii SCL2010, and the strain has been preserved in the Chinese Type Culture Collection on July 6, 2014. Center, its deposit number is CCTCC NO.M2014325. The invention also discloses the application of the sclerotin in the fermentative production of scleroglucan. By adopting the process of controlling the fermentation conditions in stages, the present invention realizes the perfect combination of rapid bacterial growth and high-efficiency sugar production, ensuring high fermentation The stable operation of the process makes the fermentation yield and production intensity reach 35g / L and 0.44g / L / h respectively, and the yield is significantly higher than that of the existing technology. The prepared product has high viscosity, good stability, and small viscosity change. Has good application performance.

Description

technical field [0001] The invention relates to a microbial polysaccharide scleroglucan production strain, in particular to a Sclerotinia sclerotiorum and its application in fermentative production of scleroglucan. Background technique [0002] Scleroglucan (also known as sclerotin or sclerotin) is a nonionic water-soluble homopolysaccharide produced by filamentous fungi. Its molecule is composed of a linear chain of β-D-(1,3)-glucose residues with side chains of β-D-(1,6)-glucose residues. The outstanding characteristics of scleroglucan are good rheology and stability, and it has good stability in a wide range of pH (1-12), salinity (0-200,000ppm) and temperature (130°C). [0003] Scleroglucan has good application value in food, medicine, cosmetics, petroleum and other industries. At present, in the international market, scleroglucan is widely used in cosmetics as moisturizing, anti-inflammatory and thickening ingredients, and is often used by internationally renowned cos...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12P19/08C12R1/645
CPCC12P19/08C12N1/145C12R2001/645
Inventor 董学前张永刚武琳吉武科刘建军王伟
Owner SHANDONG FOOD & FERMENT IND RES & DESIGN INST