Method for producing polysaccharide through liquid state fermentation of rice bran-wheat bran complete culture medium by virtue of grifola frondosa mutagenesis strain

A technology of liquid fermentation and Grifola frondosa, applied in the direction of microorganism-based methods, biochemical equipment and methods, fermentation, etc., can solve the problems of mutagenesis effect evaluation, unresolved, unsatisfactory fermentation effect, etc., and achieve advanced methods , less dosage, high economic benefits

Inactive Publication Date: 2013-10-09
JIANGSU UNIV
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Problems solved by technology

Although Guo Chunmei's master's thesis initially involved the conversion and utilization of two strains of Grifola sp.CGMCC4179 and Grifola sp.CCTCC M2011113 to the whole liquid medium of rice bran bran added with cellulase, the conclusion of the research was that the fermentation effect was not ideal , this problem has not been solved (see p57 of Guo Chunmei's master's thesis)
[0007] In 2011, Li Yongzhuan, a postgraduate student under the guidance of Liu Weimin, tried to solve the problem of liquid fermentation and utilization of rice bran and bran. He experimented with ultraviolet mutagenesis starting from Grifola sp. The rice bran and bran whole material culture medium (Master's thesis p17 rice bran and bran screening medium added agar) was solidified with agar. During the test, the mutagenic effect of the strains was not significant, so the whole rice bran and bran material could not be used. Liquid shake flask fermentation was used to evaluate the mutagenic effect, and the fermentation medium was used to filter rice bran bran juice for fermentation evaluation (Master's thesis p17 fermentation medium, similar to the experiments of Zhang Jian and Guo Chunmei), and the rice bran bran juice was taken used old road
This new strain is used in the liquid culture medium of whole material of rice bran to form a new utilization method of rice bran and realize the goal of high-yield Grifola frondosa polysaccharide. There is no literature report that can reach the production level of Grifola frondosa polysaccharide of the present invention

Method used

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  • Method for producing polysaccharide through liquid state fermentation of rice bran-wheat bran complete culture medium by virtue of grifola frondosa mutagenesis strain

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Experimental program
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Effect test

Embodiment 1

[0025] Grifola frondosa strain adopts Grifola sp.CCTCC M2013286 strain obtained by mutagenesis. The amount of rice bran used is 6.0g / 100mL medium, the amount of bran used is 2.0g / 100mL medium, and KH 2 PO 4 0.09g / 100mL, MgSO 4 0.16g / 100mL, pH is natural, add water to the required volume, sterilize at 121°C and cook for 30min, use it as a medium for fermentation, the sample volume of the shake flask is 40%, the inoculum volume is 10%, the culture temperature is 30°C, the rotation speed 180r / min, culture time 5d. The mycelium containing a small amount of rice bran bran obtained from liquid culture is centrifuged and washed with water. After the mycelium is broken, it is extracted with hot water at 70°C for 3 hours, and the clear liquid is obtained by filtration. The centrifugal separation liquid of fermented product or mycelia extracted by hot water, after deproteinization, alcohol precipitation, centrifugal separation and vacuum drying, exopolysaccharide and hyphae polysacch...

Embodiment 2

[0027] Grifola frondosa strain adopts Grifola sp.CCTCC M2013286 strain obtained by mutagenesis. The amount of rice bran used is 1.0g / 100mL medium, the amount of bran used is 1.0g / 100mL medium, and KH 2 PO 4 0.36g / 100mL, MgSO 4 0.08g / 100mL, pH is natural, add water to the required volume, sterilize at 121°C and cook for 30 minutes, use it as a medium for fermentation, the sample volume of the shake flask is 40%, the inoculum volume is 10%, the culture temperature is 25°C, the rotation speed 150r / min, culture time 7d. The mycelium containing a small amount of rice bran bran obtained from liquid culture is centrifuged and washed with water, the mycelium is broken and extracted with hot water at 70°C for 3 hours, and the clear liquid is obtained by filtration. The centrifugal separation liquid of fermented product or mycelia extracted by hot water, after deproteinization, alcohol precipitation, centrifugal separation and vacuum drying, exopolysaccharide and hyphae polysaccharid...

Embodiment 3

[0029] Grifola frondosa strain adopts Grifola sp.CCTCC M2013286 strain obtained by mutagenesis. The amount of rice bran used is 3.0g / 100mL medium, the amount of bran used is 1.0g / 100mL medium, and KH 2 PO 4 0.09g / 100mL, MgSO 4 0.16g / 100mL, natural pH, add water to the required volume, sterilize at 121°C and cook for 30 minutes, use it as a medium for fermentation, the sample volume of the shake flask is 40%, the inoculum volume is 10%, the culture temperature is 26°C, the rotation speed 150r / min, culture time 4d. The mycelium containing a small amount of rice bran bran obtained from liquid culture is centrifuged and washed with water, the mycelium is broken and extracted with hot water at 70°C for 3 hours, and the clear liquid is obtained by filtration. The centrifugal separation liquid of fermented product or mycelia extracted by hot water, after deproteinization, alcohol precipitation, centrifugal separation and vacuum drying, exopolysaccharide and hyphae polysaccharide w...

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Abstract

The invention discloses a production method of grifola frondosa polysaccharide and relates to the technical field of food microorganism application. The method comprises the following steps of: weighing raw materials, wherein rice bran is 1-6g / 100mL, wheat bran is 1-3g / 100mL, monopotassium phosphate is 0.09-0.36g / 100mL, and magnesium sulfate is 0.04-0.16g / 100mL; adding a needed amount of water into the raw materials, loading the raw materials without regulating the pH (Potential of Hydrogen), and sterilizing and boiling at 121 DEG C for 30min; after cooling, inoculating the grifola frondosa liquid seed (CCTCC (China Center For Type Culture Collection) M2013286) which is also disclosed in the invention, wherein the inoculation amount is 8-10%, the fermentation temperature is 25-30 DEG C, and the fermentation time is 4-7d; and carrying out deproteinization, alcohol precipitation, centrifugal separation and vacuum drying on the centrifugal separation liquid of a fermented product or the centrifugal separation liquid which is obtained after the mycelia which has wall broken is extracted with hot water, so as to respectively obtain exopolysaccharide and mycelia polysaccharide. By adopting the method disclosed by the invention, the purpose of efficiently converting the rice bran and the wheat bran to the grifola frondosa polysaccharide with a high value through liquid state fermentation can be achieved better.

Description

technical field [0001] The invention relates to the technical field of food microorganism application, in particular to a method for using Grifola frondosa to induce high-yield Grifola frondosa polysaccharides in liquid-state fermentation of a new strain of rice bran bran whole material medium. Background technique [0002] Since Chihara et al. (references: Chihara G, MaedaY, Hamuro J, et al. Inhibition of mouse sarcoma 180 by polysaccharides from Lentinus edodes (Berk.) sing [J]. Nature, 1969, 222 (5194): 687-688 .) published a paper on "Nature" in 1969, revealing that lentinan has anti-tumor activity, more than 200 kinds of medicinal fungi with anti-tumor activity have been found, some of which are edible fungi, such as Grifola frondosa, monkey Head fungus, Ganoderma lucidum, etc. The mycelium, fruiting bodies, sclerotia or spores of these edible fungi can produce various active ingredients such as amino acids, proteins, vitamins, polysaccharides, glycosides, flavonoids a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/04C12N13/00C12N15/01C12R1/645
Inventor 刘伟民沈国栋周文彪李亚楠江雨婷赵杰文马海乐
Owner JIANGSU UNIV
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