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Mucor racemosus SY5-47 and application thereof in extraction of flavones from mulberry leaves

A technology of Mucor racemosa and mulberry leaf powder, which is applied to medical preparations containing active ingredients, microorganism-based methods, plant raw materials, etc., can solve the problems of high cost, high equipment requirements, long time consumption, etc. , the effect of improving the extraction yield

Active Publication Date: 2021-03-30
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 yield of flavonoids also varies greatly
Such as organic solvent extraction takes a long time and the extraction yield of flavonoids is low; CO 2 Supercritical extraction and ultra-high pressure extraction can obtain higher extraction yields, but require relatively high equipment; enzyme-assisted extraction has mild extraction conditions and active substances are not easily inactivated, but the cost of enzymes is relatively high

Method used

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  • Mucor racemosus SY5-47 and application thereof in extraction of flavones from mulberry leaves
  • Mucor racemosus SY5-47 and application thereof in extraction of flavones from mulberry leaves
  • Mucor racemosus SY5-47 and application thereof in extraction of flavones from mulberry leaves

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Screening and Identification of Example 1 Bacterial Strains

[0026] 1. Screening of bacterial strains

[0027] (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 3 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.

[0028] (2) Dry heat sterilize 250-mL flask at 160°C for 2 hours, add 10 g of mulberry ...

Embodiment 2

[0042] Embodiment 2 mutagenesis selection

[0043] Carry out mutation breeding to Mucor racemos SY5, and screen the strain with excellent fermentation performance, the specific method is as follows:

[0044] (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.

[0045] (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 c...

Embodiment 3

[0048] Example 3 Mucor racemosa SY5-47 is applied to the extraction of mulberry leaf flavonoids

[0049] Mucor racemosa SY5-47 is applied to the extraction of flavonoids from mulberry leaves, and the following steps can be followed:

[0050] (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.

[0051] (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 ...

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Abstract

The invention relates to amucor racemosus SY5-47 and an application thereof in extraction of flavones from mulberry leaves. The application method comprises the following steps: adding sterile normalsaline into mulberry leaf powder, inoculating spores of the mucor racemosus SY5-47, fermenting at 28-30 DEG C for 3-4 days, carrying out ultrasonic ethanol extraction on the fermented product, concentrating to obtain a concentrate, and carrying out vacuum drying to obtain the mulberry leaf flavone extract. Before flavone is extracted from mulberry leaves by ultrasonic ethanol, fermentation pretreatment of mucor racemosus SY5-47 is added. The mucor racemosus SY5-47 is an excellent strain obtained by screening from a mulberry leaf microorganism enrichment culture and mutation breeding, polysaccharide hydrolase is generated by moderate growth in mulberry leaf powder, mulberry leaf cell walls are hydrolyzed, so that mulberry leaf structure tissues are loose, flavonoid compounds are easier to dissolve out during ultrasonic ethanol extraction, compared with an ultrasonic ethanol extraction method without fermentation pretreatment, the method has the advantage that the extraction yield of themulberry leaf flavone can be increased by nearly 50%.

Description

[0001] (1) Technical field [0002] The invention belongs to the technical field of biochemical industry, and in particular relates to the application of a microbial fermentation technology in the extraction of flavonoids from mulberry leaves. [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 and headache, red eyes and blurred vision. 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 flavonoids are one of the main active ingredients. [0005] Morus leaf flavonoids have significan...

Claims

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Application Information

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IPC IPC(8): C12N1/14A61K36/605A61K127/00C12R1/785
CPCA61K36/605A61K2236/19A61K2236/333A61K2236/51
Inventor 梅建凤庄仕航季军情易喻王旭东应国清
Owner ZHEJIANG UNIV OF TECH
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