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Method of preparing bioactive substance with co-cultivation of plant-based raw material and endophytic bacteria

A technology of biologically active substances and endophytic bacteria, applied in the biological field, can solve the problems of easy degeneration of bacterial strains, and achieve the effects of short production cycle, controllable preparation process and stable level

Inactive Publication Date: 2017-07-04
NANJING BIOFUNCTION CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

As introduced in the article "Discussion on the Degeneration of Cordyceps Militaris", Cordyceps militaris has very similar pharmacological effects to Cordyceps sinensis and can be used as a substitute for Cordyceps sinensis, but its strains are easily degraded during subculture

Method used

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  • Method of preparing bioactive substance with co-cultivation of plant-based raw material and endophytic bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The method for preparing biologically active substances by co-cultivating plant-based raw materials and endophytes specifically includes the following steps:

[0030] (1) Pretreatment of plant-based raw materials: plant-based raw materials select gardenia buds as explants, and obtain sterile gardenia explants by treating with mercuric chloride, alcohol and sterile water, and cultivate gardenia callus The formula of the culture medium of Gardenia jasminoides callus is: adopt MS liquid as basic culture liquid, add hormone 2,4-D 4.0mg / L, 6-BA 2.0mg / L, sucrose concentration 10g / L, agar concentration 1.5 g / L, adjust the pH value to 5.8; set the culture conditions as follows: culture in the dark for 20 days, then transfer to culture under light conditions, culture time is 15 days, light intensity is 1800lx, light time is 18h / d, and the temperature is 24±1°C. After the callus grows, it is transferred to the callus solid proliferation step, and the solid medium formula is: adop...

Embodiment 2

[0036] The method for preparing biologically active substances by co-cultivating plant-based raw materials and endophytes specifically includes the following steps:

[0037] (1) Pretreatment of plant-based raw materials: select yew explants as plant-based raw materials, obtain sterile yew explants by treating with mercuric chloride and sterile water, and cultivate yew callus; yew callus The formula of the tissue culture medium is: use MS liquid as the basic culture medium, add hormone 2,4-D 2.0mg / L, 6-BA 1.0mg / L, sucrose concentration 30g / L, agar concentration 6.0g / L, adjust The pH value is 6.0; the culture conditions are set as follows: culture in the dark for 25 days, then transfer to culture under light conditions, culture time is 13 days, light intensity is 1800lx, light time is 18h / d, and temperature is 24±1°C. After the callus grows, it is transferred to the callus solid proliferation step, and the solid medium formula is: adopt MS liquid as the basic culture solution, a...

Embodiment 3

[0043] The method for preparing biologically active substances by co-cultivating plant-based raw materials and endophytes specifically includes the following steps:

[0044] (1) Pretreatment of plant-based raw materials: select yew leaf extracts as plant-based raw materials, and obtain sterile yew leaf extracts through high-pressure moist heat sterilization;

[0045] (2) Activation of endophytes: the endophytes of Taxus chinensis were inoculated on the microbial medium containing different biological material concentration gradients for pre-cultivation, and the pre-cultivation time was 5 days, so as to obtain activated endophytes;

[0046] (3) Co-cultivation of biological materials and endophytes: the yew leaf extract obtained in step (1) and the endophytes isolated from yew Phomopsis TCN-01 co-culture, the co-culture conditions are: 1) Phomopsis TCN-01 was cultured on ordinary PDA solid medium for 5 days, and the culture temperature was 25°C; 2) Potato dextrose liquid (P...

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Abstract

The invention relates to a method of preparing a bioactive substance with co-cultivation of a plant-based raw material and endophytic bacteria. The method includes the steps of: 1) pretreatment to the plant-based raw material: preparing sterile-cultured propagules, such as living plant cells and tissues, or drying and / or performing pulverization to the plant-based raw material, and performing sterilization pretreatment; 2) activation to the endophytic bacteria: inoculating the endophytic bacteria on a microbial culture medium to perform pre-cultivation for 1-25 days to produce activated endophytic bacteria; 3) co-cultivation of the bio-material and the endophytic bacteria: in a co-cultivation device, inoculating the activated endophytic bacteria in the step 2) and adding a solid or liquid culture medium, which contains the plant-based raw material and is prepared in the step 1), to perform mixed cultivation for 4-90 days to obtain a co-cultivation product; and 4) extraction and preparation of the bioactive substance. Through the co-cultivation of the plant-based raw material and the endophytic bacteria, the bioactive substance is prepared. The method is simple and controllable, achieves stable expression of the product and is short in production period.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for co-cultivating biologically active substances with plant-based raw materials and endophytes. Background technique [0002] Bioactive substances refer to a type of trace or small amount of substances that come from living organisms and have a regulatory effect on life processes. They have the following characteristics: first, low content, and second, high biological activity. Bioactive substances with medicinal uses refer to a class of natural components extracted from biological tissues or biomass that have significant regulatory effects on humans or animals. These natural components especially have a positive effect on improving human health. Such biologically active substances have become the goals pursued by related industries through extraction, modification, modification and transformation. [0003] The biological groups in nature vary widely, and each organism co...

Claims

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Application Information

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IPC IPC(8): C12P1/02C12P1/04C12P1/06
CPCC12P1/02C12P1/04C12P1/06Y02P20/54
Inventor 陈集双陈国晶樊小宽王炜张保钱金磊磊张本厚胡燕花
Owner NANJING BIOFUNCTION CO LTD
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