Method for producing Wulingshen exopolysaccharide by deep liquid fermentation

A technology of submerged liquid fermentation and exopolysaccharides, applied in the field of microorganisms, can solve the limitations, no research to solve the problems of medium formula selection, fermentation operation parameters control polysaccharide extraction conditions, etc., and achieve the effect of solving technical problems

Active Publication Date: 2016-06-15
JIANGSU SHENHUA PHARMA
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Problems solved by technology

At present, there are only a few colleges and universities in China to carry out research on the production of Wulingshen by deep liquid fermentation, and most of them are limited to the level of small experiments. They are all researched with the main goal of obtaining the mycelium of Wulingshen. There are almost no relevant reports on the production of functional component polysaccharides of Wuling ginseng by liquid fermentation, and there is no research to solve technical problems such as the selection of medium formula, control of fermentation operating parameters, and research on polysaccharide extraction conditions encountered in the process of enlarging from small-scale grafting to industrial production. , so at present there is no real industrialized liquid fermentation production of Wuling ginseng polysaccharide in China

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  • Method for producing Wulingshen exopolysaccharide by deep liquid fermentation

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Experimental program
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Effect test

Embodiment 1

[0033] The method for producing exopolysaccharide of Wulingshen by industrialized submerged liquid fermentation of the present invention is carried out according to the following steps:

[0034]1. Preparation of Wulingshen fermentation broth: Inoculate the activated Wulingshen shake flask strain into seed medium (300L) according to the inoculum size 1%, wherein the seed medium consists of 2% glucose, 0.2% peptone, soybean cake Powder 1%, potassium dihydrogen phosphate 0.3%, magnesium sulfate heptahydrate 0.15%, zinc sulfate 0.10%, soybean oil 0.02%, adjust the pH value to 5.5, ventilation ratio 1:0.3vvm, tank pressure 0.03Mpa, oscillate at 26-28°C Cultivate for 4 days to obtain the first-grade seed liquid of Wulingshen; insert the first-grade seed liquid into the secondary seed medium (3000L) according to the inoculum size of 15%, wherein the secondary seed medium consists of 2% glucose, 0.2% peptone, Soybean cake powder 1%, potassium dihydrogen phosphate 0.3%, magnesium sulfa...

Embodiment 2

[0039] 1. Preparation of Wulingshen fermentation broth: Inoculate the activated Wulingshen shake flask strain into seed medium (300L) according to the inoculum size 1.5%, wherein the seed medium consists of glucose 2%, peptone 0.2%, yeast powder 1%, potassium dihydrogen phosphate 0.3%, magnesium sulfate heptahydrate 0.15%, zinc sulfate 0.10%, rapeseed oil 0.02%, adjust the pH value to 5.5, the ventilation ratio is 1:0.3vvm, the tank pressure is 0.03Mpa, 26~28℃ Shake culture for 3 days to obtain the first-class seed liquid of Wulingshen; the first-class seed liquid was inserted into the second-level seed medium (3000L) according to the inoculation amount of 15%, and the second-level seed medium was composed of 2% glucose and 0.2% peptone , soybean cake powder 1%, potassium dihydrogen phosphate 0.3%, magnesium sulfate heptahydrate 0.15%, zinc sulfate 0.10%, defoamer 0.02%, adjust the pH value to 5.5, the ventilation ratio is 1:0.3vvm, the tank pressure is 0.03Mpa, Cultivate at 2...

Embodiment 3

[0044] In order to better verify the superiority of the composite prescription of short-acting glucose and long-acting starch in the industrial production of Wulingshen polysaccharide in the present invention, several comparative experiments were done. Only adjusted the consumption of carbon source glucose or (and) starch in the test in the liquid fermentation medium, do not contain starch in the comparative example 1, and glucose concentration is 1.5%; Do not contain glucose in the comparative example 2, and the starch concentration is 2.5%, to In ratio 3, the glucose content is 2.5%, and the starch content is 0.5%; all the other conditions are the same as the present invention, and the test results are shown in Table 1:

[0045] Table 1

[0046]

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Abstract

The invention discloses a method for producing Wulingshen exopolysaccharide by submerged liquid fermentation. The method is to expand the activated Wuling ginseng strain by inserting it into the seed tank, and then insert the expanded liquid seed into a special polysaccharide-producing liquid medium for fermentation, and then ferment the fermentation liquid. Separation and extraction of exopolysaccharides. The present invention is aimed at the technical difficulties encountered in the scale-up of the small scale test to the industrialized production. On the basis of continuing to optimize the fermentation medium (especially the carbon source) and increasing the biomass and exopolysaccharides in the small test and pilot scale , solved the problems of fermentation process parameter control, extraction equipment selection and optimization encountered in the enlargement process, and finally successfully grafted into a 10-20t fermenter for industrial production, and successfully produced high-quality Wulingshen exopolysaccharide , truly realized the continuous, automatic and industrialized production.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a method for producing Wulingshen exopolysaccharide by submerged liquid fermentation. Background technique [0002] Wulingshen belongs to the class Ascomycetes, and is a traditional medicinal fungus. It grows on the abandoned nests of black-winged earth-dwelling termites deep underground. It is regarded as a rare and valuable medicinal material by the research and application fields of medicinal fungi in the world, and has extensive medicinal value. [0003] Wuling ginseng grows on sunny and warm hill slopes or termite waste fungi on river embankment soil slopes. It is densely covered with mycelia of C. nigriscens on the fungus garden. In the later stage of its growth, part of the sclerotium is densely formed. , commonly known as "Wuling Ginseng", weighing about 5-30 grams, spherical, spherical or irregular flat strips, 2-12 cm in diameter, brown or black in a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/04C08B37/00C12R1/645
Inventor 王文风龚劲松徐国华王英燕张芙蓉杨亚威高岩
Owner JIANGSU SHENHUA PHARMA
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