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Method for extracting folium cortex eucommiae polysaccharide by utilizing microbial fermentation method

A microbial fermentation method and the technology of Eucommia leaves, applied in the field of microbial fermentation technology in the extraction of polysaccharides from Eucommia leaves, can solve the problems of increasing production costs and limiting industrial applications, and achieve the promotion of dissociation, easy dissolution, and improved extraction yield Effect

Pending Publication Date: 2021-10-29
ZHEJIANG SHUREN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although enzyme-assisted extraction can increase the yield of polysaccharides, the use of commercial pure enzymes undoubtedly increases production costs and limits its industrial application

Method used

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  • Method for extracting folium cortex eucommiae polysaccharide by utilizing microbial fermentation method
  • Method for extracting folium cortex eucommiae polysaccharide by utilizing microbial fermentation method
  • Method for extracting folium cortex eucommiae polysaccharide by utilizing microbial fermentation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Isolation and screening of fermentation strains

[0021] The microbial strains fermenting Eucommia leaves were isolated and screened according to the following steps:

[0022] (1) About 5 g of eucommia leaf powder was added to a 250-mL Erlenmeyer flask, and then 5 mL of sterile saline was added to moisten it, and incubated at a constant temperature of 28°C for 4 days. The enriched cultures covered with mold were diluted 1×10 with sterile saline -6 , 1×10 -7 , 1×10 -8 After doubling, draw 0.1mL of the diluted solution and spread it on the potato dextrose agar plate medium (PDA), culture at a constant temperature of 28°C for 2 days, pick mold colonies with different colors and shapes and transfer them to fresh PDA plate medium, and place them at 28°C. After culturing at constant temperature at ℃ for 3 days, 14 purely cultured strains were obtained, and the numbers of each strain are shown in Table 1.

[0023] (2) Add 5 mL of sterile saline to the fresh plat...

Embodiment 2

[0034] Embodiment 2: the mutagenesis selection of fermentation strain DZ-9

[0035] Carry out mutagenesis breeding on strain DZ-9, and screen strains with excellent fermentation performance, the specific method is as follows:

[0036] (1) Preparation of spore liquid: After the bacterial strain DZ-9 was activated and cultured on PDA plate medium for 2 days, 5 mL of sterile normal saline was added, stirred with an inoculation loop to suspend the spores, and then transferred to a triangular flask containing 45 mL of sterile normal saline Medium (with 20–30 glass beads added), shake at room temperature for 20 minutes. The spore suspension was filtered to remove mycelium (triangular funnel pad with 2 layers of lens-cleaning paper), counted the spores in the suspension with a hemocytometer under a microscope, and diluted it with sterile saline to adjust the number of spores to 1× 10 8 individual / mL.

[0037] (2) Mutagenesis: under red light illumination, take 2.0mL of the above s...

Embodiment 3

[0039] Example 3: Classification and Identification of Bacterial Strain DZ-9-67

[0040] Strain DZ-9-67 was inoculated on PDA plate medium and cultured at 28°C for 3 days. The colonies were grayish white and short fluffy at the beginning, and then gradually turned green to olive green. Sporulation. The top of the aerial hyphae produces conidiophores, with 1–4 branches at the top of the stalk, and the terminal cells of each branch divide into clusters of conidia, forming a typical brush-shaped conidia spike; the conidia are spherical or Nearly spherical, 2.5–3.0 μm in diameter, green. See the photos of the colony of Penicillium papaya DZ-9-67 cultured on PDA plate medium at 28°C for 3 days figure 1 .

[0041] The strain DZ-9-67 was handed over to Sangon Bioengineering (Shanghai) Co., Ltd. and the rDNA nucleotide sequence of its ribosomal ITS region was measured as SEQ ID NO.1, which is listed in NCBI (National Center for BiotechnologyInformation, https : / / www.ncbi.nlm.nih.g...

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Abstract

The invention discloses a method for extracting folium cortex eucommiae polysaccharide by using a microbial fermentation method, which comprises the following steps: adding a sterile cane sugar aqueous solution into folium cortex eucommiae powder, inoculating penicillium velutinum DZ-9-67 spores, fermenting at 28-30 DEG C for 2-3 days, adding deionized water into the fermented product to extract at 25-30 DEG C, then carrying out ultrasonic-assisted extraction, concentrating to 1 / 10-1 / 7.5 of the original volume to obtain a concentrate, adding an ethanol aqueous solution into the concentrate, precipitating at 4 DEG C, centrifuging, and drying the precipitate in vacuum to obtain a crude extract of the folium cortex eucommiae polysaccharide. the fermentation pretreatment of the penicillium velutinum DZ-9-67 is added in advance of ultrasonic water extraction of the folium cortex eucommiae polysaccharide, and compared with an ultrasonic water extraction method without fermentation pretreatment, the method has the advantages that the extraction yield of the folium cortex eucommiae polysaccharide can be increased by 33.7%.

Description

(1) Technical field [0001] 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 eucommia leaf polysaccharide. (2) Background technology [0002] Eucommia ulmoides Oliv., also known as Bakelite, is a plant of the genus Eucommia in the family Eucommia. Eucommia has been cultivated in my country for more than two thousand years. The first medicinal book in my country, "Shen Nong's Materia Medica" in the Han Dynasty, recorded the efficacy of Eucommia bark, and listed Eucommia as top grade. Li Shizhen's book "Compendium of Materia Medica" in the Ming Dynasty described the origin and efficacy of Eucommia in more detail. Eucommiae Cortex and Eucommiae Folium are included in the 2020 edition of the Chinese Pharmacopoeia, a medicinal material and decoction pieces. The pharmacopoeia records the functions and indications of Eucommia ulmoides as: invigorating live...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/04C12P1/02C12R1/80
CPCC12P19/04C12P1/02
Inventor 陈虹张建芬雷超柯薇陆胤
Owner ZHEJIANG SHUREN UNIV
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