Endophytic fungus originated from pigeon pea and capable of efficiently converting ginsenoside Rb1 into ginsenoside Rd and application thereof

A technology of ginsenosides and endophytic fungi, which is applied in the field of microbial technology applications, can solve problems such as high energy consumption, low content, and environmental pollution, and achieve high application value and overcome the effect of low content

Active Publication Date: 2017-11-03
NORTHEAST FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The invention overcomes the disadvantages of low content, high energy consumption, and environmental po...

Method used

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  • Endophytic fungus originated from pigeon pea and capable of efficiently converting ginsenoside Rb1 into ginsenoside Rd and application thereof
  • Endophytic fungus originated from pigeon pea and capable of efficiently converting ginsenoside Rb1 into ginsenoside Rd and application thereof
  • Endophytic fungus originated from pigeon pea and capable of efficiently converting ginsenoside Rb1 into ginsenoside Rd and application thereof

Examples

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Embodiment 1

[0015] Example 1: Isolation of Fusarium proliferatum G11-7 in the present invention.

[0016] In this embodiment, Fusarium proliferatum G11-7 is isolated from healthy pigeonpea roots. The pigeonpea roots are collected from the botanical garden of Northeast Forestry University, and the plant is 3 months old. The samples are currently stored in Northeast Forestry University. Key Laboratory of Forest Plant Ecology Ministry of Education; it is isolated and cultivated according to the following steps:

[0017] (1) Preparation of Potato Dextrose Agar Medium (PDA): Select mature, smooth surface, no buds and no ulcerated potatoes, wash and peel them, weigh 200g of potatoes and cut them into pieces, boil them in boiling water for 30min, and filter them through 4 layers of gauze to obtain Add 20g of glucose and 20g of agar powder to the potato liquid, add distilled water to make it up to 1L, keep the pH natural, and sterilize under high temperature and high pressure at 121°C for 15 minu...

Embodiment 2

[0023] Example 2: Screening of Fusarium proliferatum G11-7 strain directed transformation of notoginseng ginsenoside Rb1 to Rd in the present invention.

[0024] (1) Preparation of screening medium: cellulose-Congo red solid medium contains 0.2% sodium carboxymethylcellulose, 0.2% (NH 4 ) 2 S0 4 , 0.05% MgSO 4 ·7H 2 O, 0.1% KH 2 P0 4 , 1.5% agar, adjust the pH to 7.0, and sterilize under high temperature and high pressure at 121°C for 15 minutes;

[0025] (2) Inoculate the pigeon pea endophytic fungus strain into the cellulose-Congo red solid medium, cultivate until regular colonies grow, and cover the colonies with a Congo red solution with a mass concentration of 1 mg / mL for 10 to 15 minutes Finally, pour off Congo red solution, add the NaCl solution that the amount concentration of substance is 1mol / L, pour off NaCl solution after 15min, at this moment, there will be transparent circle around the bacterium colony that produces cellulase, the diameter of transparent ci...

Embodiment 3

[0027] Example 3: Direct transformation of Panax notoginsenoside Rb1 into Rd by Fusarium proliferatum G11-7 strain of the present invention.

[0028] (1) Strain activation: the pigeonpea endophytic fungus Fusarium proliferatum (Fusarium proliferatum) G11-7 was inoculated on the PDA solid slant medium and cultivated for 7-10 days;

[0029] (2) Preparation of transformation medium: select mature, smooth surface, no buds and no ulcerated potatoes, wash and peel them, weigh 200g potatoes and cut them into pieces, boil them in boiling water for 30min, and dilute the potato liquid obtained after filtering through 4 layers of gauze to 1L, pH natural;

[0030] (3) Pretreatment of the raw powder of the notoginseng medicinal material: crush the raw material of the notoginseng medicinal material and pass through a 40-mesh sieve to obtain the raw medicinal material powder of the notoginseng medicinal material. Bacteria 40min;

[0031] (4) Fermentation transformation: take the slant stra...

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Abstract

The invention discloses an endophytic fungus strain, i.e., Fusarium proliferatum G11-7, originated from a pigeon pea and capable of efficiently converting ginsenoside Rb1 into ginsenoside Rd and a separation method thereof, and application of the strain to fermentation conversion of ginsenoside. The strain G11-7 is originated from a pigeon pea plant and prepared through solid culture, separation and purification. The strain can efficiently and specifically convert ginsenoside Rb1 into ginsenoside Rd, and the content of ginsenoside Rd after conversion is about 4 times the content of ginsenoside Rd before conversion. The Fusarium proliferatum G11-7 provided by the invention is an endophytic fungus acquired from the pigeon pea for the first time. When the Fusarium proliferatum G11-7 is applied to conversion of ginsenoside Rb1 into ginsenoside Rd, conversion time is short, conversion rate is high, no pollution is produced, industrial production can be easily realized, and good application prospects are obtained.

Description

technical field [0001] The invention relates to an endophytic fungus capable of efficiently transforming ginsenoside Rb1 into Rd and its application, belonging to the field of microbial technology applications. technical background [0002] Endophytic fungi refer to fungi that live in plant tissues at a certain stage of their life history, but do not cause obvious plant disease symptoms. Through "co-evolution" with medicinal plants, some endophytic fungi have the ability to produce the same or similar biologically active substances as the host plant (Chen Lijun et al., 2006). Endophytic fungi have rich species diversity, and their secondary metabolites are also very rich. A series of enzymes can be produced through the vegetative culture and fermentation culture of endophytic fungi, and these microbial-derived enzymes can interact with the substrate to change its structure. Therefore, microbial transformation of endophytic fungi has the ability to It has the characteristic...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P33/20C12R1/77
CPCC12P33/20C12N1/145C12R2001/77
Inventor 付玉杰牛丽丽盖庆岩焦骄郭娜王希清王微
Owner NORTHEAST FORESTRY UNIVERSITY
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