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Method for extracting mulberry leaf polysaccharide by utilizing microbial fermentation method

A microbial fermentation method and the technology of mulberry leaf polysaccharide, which are applied in the directions of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of increased production cost, long time consumption, low polysaccharide extraction rate, etc., to promote dissociation, The effect of easy dissolution and improved extraction yield

Active Publication Date: 2021-03-12
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Different extraction processes have their own advantages and disadvantages, and the extraction yield of polysaccharides also varies greatly
The hot water extraction method takes a long time and the extraction rate of polysaccharides is low; the ultrasonic or microwave-assisted extraction method is based on hot water extraction, adding ultrasonic or microwave treatment steps, and the extraction yield is relatively high, so it has relative advantages; The enzyme-assisted extraction method is based on the previous three extraction methods, adding an enzyme treatment step. Although a higher yield of polysaccharides can be obtained, the use of enzymes undoubtedly increases production costs and limits its wide application.

Method used

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  • Method for extracting mulberry leaf polysaccharide by utilizing microbial fermentation method
  • Method for extracting mulberry leaf polysaccharide by utilizing microbial fermentation method
  • Method for extracting mulberry leaf polysaccharide by utilizing microbial fermentation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Screening and identification of embodiment 1 fermentation strain

[0024] 1. Strain screening

[0025] (1) Add about 10 g of mulberry leaf powder to a 250-mL Erlenmeyer flask, then add 25 mL of sterile saline to moisten it, and incubate at a constant temperature of 28°C for 3 days. Dilute the mold-covered enrichment culture 1×10 with sterile saline 8 After doubling, spread it on the PDA plate medium, culture at 28°C for 2 days, pick out mold colonies with different colors and shapes, transfer them to fresh PDA medium, and place them in 28°C for 2 days to obtain 8 purely cultured strains. They were numbered SY1–SY8. Add 5 mL of sterile saline to the fresh plate cultures of the 8 strains, stir with an inoculation loop to suspend the spores, transfer the spore liquid to a sterile test tube, and adjust the concentration of spores with sterile saline to 3×10 8 individual / mL.

[0026] (2) Dry heat sterilize 250-mL flask at 160°C for 2 hours, add 10 g of mulberry leaf powd...

Embodiment 2

[0041] Embodiment 2 mutation breeding

[0042] Carry out mutation breeding to Mucor racemos SY5, and screen the strain with excellent fermentation performance, the specific method is as follows:

[0043] (1) Preparation of spore liquid: After mucormyces racemosa SY5 is activated and cultivated by PDA, add 5 mL of sterile normal saline, stir with an inoculation loop to suspend the spores, then transfer to a triangular flask containing 45 mL of sterile normal saline, at room temperature Shake for 30min. The spore suspension was filtered to remove mycelia (a cotton ball was placed on the neck of the triangular funnel to filter), and the spores in the suspension were counted with a hemocytometer under a microscope, and diluted with sterile saline to adjust the number of spores to 1 ×10 8 A / mL or so.

[0044] (2) Mutagenesis: mutagenesis was performed under red light. Take 2.0 mL of the above spore suspension and a sterile paper clip into a petri dish with a diameter of 6 cm, p...

Embodiment 3

[0047] Example 3 Mucor racemosa SY5-47 is applied to the extraction of mulberry leaf polysaccharides

[0048] (1) Inoculate Mucor raceme SY5-47 (freeze-dried tube or plate colony) preserved at low temperature on fresh PDA plate medium, culture at a constant temperature of 28°C for 2 days, add 5 mL of sterile saline to the Petri dish, and inoculate with Ring agitation to suspend the spores, transfer the spore liquid to a sterile test tube, and adjust the spore concentration to 3×10 with sterile saline. 8 individual / mL. Described PDA plate medium composition and preparation method are the same as embodiment 1.

[0049] (2) 250-mL Erlenmeyer flask was sterilized by dry heat at 160°C for 2 hours, 10 g of mulberry leaf powder was added, 25 mL of sterile saline was added, and 1 mL of the spore liquid of Mucor racemosus SY5-47 prepared in step (1) was inoculated. , stir well. The mouth of the triangular flask was tied with eight layers of gauze, and cultured at 28°C for 3 days to ...

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Abstract

The invention discloses a method for extracting mulberry leaf polysaccharide by using a microbial fermentation method, which comprises the following steps: adding sterile normal saline into mulberry leaf powder, inoculating spores of Mucor racemosum SY5-47, fermenting at 28-30 DEG C for 3-4 days, carrying out ultrasonic water extraction on the fermented product, and concentrating the extract to obtain a concentrate; and carrying out alcohol precipitation and vacuum drying on the concentrate to obtain mulberry leaf crude polysaccharide, wherein a step of the fermentation pretreatment of mucor racemosus SY5-47 is carried out before ultrasonic water treatment of mulberry leaf polysaccharide. According to the technical scheme, the Mucor racemosus SY5-47 grows moderately in mulberry leaf powderto generate various polysaccharide hydrolases to hydrolyze cell walls of mulberry leaves, so that structural tissues of the mulberry leaves are loose, dissociation of combined soluble polysaccharidesis promoted, and the polysaccharide hydrolases are easier to dissolve out during hot water ultrasonic extraction. Compared with an ultrasonic water extraction method without fermentation pretreatment, the extraction yield of the mulberry leaf polysaccharide can be increased by 35% or above.

Description

[0001] (1) Technical field [0002] The invention belongs to the field of biochemical technology, and in particular relates to the application of a microbial fermentation technology in the extraction of mulberry leaf polysaccharides. [0003] (2) Background technology [0004] Mulberry leaf is the leaf of Morus alba L., a plant of the Moraceae family. It is a traditional Chinese medicinal material included in the "Chinese Pharmacopoeia". Dry cough, dizziness, headache, blurry eyes. Modern pharmacological studies have shown that mulberry leaves have pharmacological effects such as anticoagulation, lowering blood pressure, lowering blood sugar, lowering blood fat, anti-aging, and anti-tumor. The chemical components of mulberry leaves are polysaccharides, flavonoids, alkaloids, vitamins, carotene and many amino acids needed by the human body, among which polysaccharides are one of the main active ingredients. [0005] Modern medical research shows that mulberry leaf polysacchari...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P19/04C08B37/00C12R1/785
CPCC12N1/14C12P19/04C08B37/0003
Inventor 梅建凤季军情庄仕航易喻王旭东应国清
Owner ZHEJIANG UNIV OF TECH
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