Rhizopus oryzae CH5 and application thereof in extracting ganoderma sinensis polysaccharide

A technology of Rhizopus oryzae and Zizhi powder is applied in the field of bioengineering, which can solve the problems of reduced physiological activity and damage of polysaccharide structure, and achieves the effect of promoting dissociation and improving extraction yield.

Active Publication Date: 2020-06-26
ZHEJIANG SHUREN COLLEGE ZHEJIANG SHUREN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although some advanced technologies have greatly improved the yield of polysaccharides, so

Method used

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  • Rhizopus oryzae CH5 and application thereof in extracting ganoderma sinensis polysaccharide
  • Rhizopus oryzae CH5 and application thereof in extracting ganoderma sinensis polysaccharide
  • Rhizopus oryzae CH5 and application thereof in extracting ganoderma sinensis polysaccharide

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Isolation, screening and identification of embodiment 1 fermentation strain

[0028] 1. Isolation and screening of strain CH5

[0029] (1) Add 20g of Zizhi powder crushed through a 40-mesh sieve into a 250-mL Erlenmeyer flask, then add 30mL of sterile water to moisten it, and incubate at 28°C for 4 days. Dilute the enriched culture covered with mold with sterile water 1 × 10 6 After doubling, spread it on the PDA plate medium, and culture it at 28°C for 3 days, pick out mold colonies with different colors and shapes and transfer them to fresh PDA medium, and place them in 28°C constant temperature culture for 3 days to obtain 7 purely cultured strains, respectively Numbered CH1–CH7. Add 5 mL of sterile water to the fresh plate cultures of the 7 strains, stir with an inoculation loop to suspend the spores, transfer the spore liquid to a sterile test tube, and adjust the spore concentration to 3× with sterile water. 10 8 individual / mL.

[0030] The plate medium is pot...

Embodiment 2

[0042] Embodiment 2 Rhizopus oryzae CH5 is applied to the extraction of Zizhi polysaccharide

[0043] (1) Preparation of Rhizopus oryzae CH5 spore liquid: Plate colonies of Rhizopus oryzae CH5 preserved in a refrigerator at 4°C were inoculated on fresh PDA plate medium, cultivated at a constant temperature of 28°C for 3 days, and added 5 mL of sterile water to the plate culture, Stir with an inoculation loop to suspend the spores, transfer the spore liquid to a sterile test tube, and adjust the spore concentration to 4×10 with sterile water. 8 Individual / mL; Described PDA plate medium composition and preparation method are with embodiment 1;

[0044] (2) 500-mL triangular flask was sterilized by dry heat at 160°C for 2 hours, added 20 g of Zizhi powder crushed through a 40-mesh sieve, added 30 mL of sterile water, and then inoculated with 2 mL of Rhizopus oryzae CH5 spore suspension prepared in step (1). liquid, stir well. The mouth of the triangular flask was tied with eigh...

Embodiment 3

[0049] Embodiment 3 Rhizopus oryzae CH5 is applied to the extraction of Zizhi polysaccharide

[0050] (1) Preparation of Rhizopus oryzae CH5 spore liquid: cryopreserved Rhizopus oryzae CH5 (freeze-dried tube or plate colony) was inoculated on fresh PDA plate medium, cultured at 28°C for 3 days, and 5 mL of sterile water was added to the plate In the culture, agitate with an inoculation loop to suspend the spores, transfer the spore liquid to a sterile test tube, and adjust the spore concentration to 5×10 with sterile water. 8 Individual / mL; Described PDA plate medium composition and preparation method are with embodiment 1;

[0051] (2) The 2-L triangular flask was sterilized by dry heat at 160°C for 2 hours, added 50g of Zizhi powder crushed through a 40-mesh sieve, added 100mL of sterile water, and then inoculated with 5mL of Rhizopus oryzae CH5 spore suspension liquid, stir well. The mouth of the triangular flask was tied with eight layers of gauze, cultured at 30°C for 2...

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Abstract

The invention discloses rhizopus oryzae CH5 and application thereof in extracting ganoderma sinensis polysaccharide. The application method comprises the following steps of: inoculating rhizopus oryzae CH5 spores into ganoderma sinensis powder containing sterile water, fermenting at 28-30 DEG C for 2-3 days, and drying the fermented product to obtain fermented ganoderma sinensis powder; decolorizing the fermented ganoderma sinensis powder to obtain decolorized ganoderma sinensis powder; performing water extraction and concentration on the decolorized ganoderma sinensis powder to obtain a concentrated solution; and performing alcohol precipitation and vacuum drying on the concentrated solution to obtain ganoderma sinensis crude polysaccharide. The rhizopus oryzae CH5 is separated from a microbial enrichment culture of ganoderma sinensis, and a screened excellent strain moderately grows in the ganoderma sinensis powder to generate polysaccharide hydrolase, so that hydrolysis of insolublepolysaccharide structural tissues of ganoderma sinensis is realized, dissociation of combined soluble polysaccharide is promoted, and the extraction yield of ganoderma sinensis polysaccharide is remarkably increased. Under the optimal process conditions, the extraction yield of ganoderma sinensis polysaccharide is 5.92 percent, which is increased by over 35 percent compared with that of an unfermented extraction method.

Description

[0001] (1) Technical field [0002] The invention belongs to the technical field of bioengineering, and in particular relates to a method for improving the extraction yield of polysaccharides from Zizhi lucidum by adopting a microbial fermentation method. [0003] (2) Background technology [0004] Ganoderma sinense (Ganoderma sinense), also known as Heizhi and Xuanzhi, also known as "Chinese Ganoderma lucidum" and "Ganoderma sinense", is a fungus of the genus Ganoderma lucidum belonging to the family Polyporaceae. Zizhi is a kind of famous and precious traditional Chinese medicine Ganoderma lucidum in my country. It is recorded in "Compendium of Materia Medica": Zizhi is a woody fungus, with a sweet, warm and non-toxic smell; Take it for a long time, lighten your body and prolong your life. Ganoderma lucidum, which is recorded in the first part of the Chinese Pharmacopoeia in the 2015 edition, is a kind of Ganoderma lucidum. It has the functions of invigorating qi and calming...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P19/04C08B37/00C12R1/845
CPCC12P19/04C08B37/0003C12R2001/845C12N1/145
Inventor 陈虹张建芬雷超谢广发
Owner ZHEJIANG SHUREN COLLEGE ZHEJIANG SHUREN UNIV
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