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A hydrogel-immobilized microbial preservation method

A technology of microorganisms and hydrogels, which is applied in the field of hydrogel immobilized microorganisms preservation, can solve the problems of insufficient research on detection indicators, type control of antifreeze protection agents, inability to be widely used in production, limited number of control experiments, etc., and achieve the goal of maintaining microorganisms Vitality, prevention of cell damage, and ease of operation

Active Publication Date: 2022-12-02
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Our country started relatively late in the field of strain preservation technology research, and has not yet formed a systematic operating procedure. The main problems are: the number of controlled tests is limited, the types of tests are small, and the research technical means need to be further updated. Insufficient research on the control of bacterial age, size of inoculated mycelium block, medium type, speed of cooling and freezing and warming and recovery, detection indicators, type of antifreeze protectant, etc.)
However, there are still problems. For example, even for the same species of Cordyceps militaris, different researchers use similar methods and draw different conclusions.
At the same time, liquid nitrogen cryopreservation and vacuum freeze-drying storage methods require good equipment and technology, and the investment is too high to be widely used in production; although the latter two methods are simple and cheap, they occupy a large storage space, and due to Too many tube transfers will increase the possibility of variation, which will easily cause strain degradation and bring losses to production

Method used

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  • A hydrogel-immobilized microbial preservation method
  • A hydrogel-immobilized microbial preservation method
  • A hydrogel-immobilized microbial preservation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Preparation of a composite gel ball:

[0034] S1: Add the chitosan powder to the acetate solution (concentration of 2.1 %, w / w), stir until completely dissolved; add CACO under the magnetic mixing 3 (The mass ratio of the chitosan powder is 0.79: 1), stir for 12min, and then the ultrasonic 15min; then add liquid paraffin (the solid liquid to the alpine glycogen powder and the paraffin is 1g: 156ml), stir 5min violently; 43 ℃ water bath water bath; Under the conditions, add the SPAN-80 (the solid ratio of the chitosan powder and the SPAN-80 is 1g: 2.1ml), stir for 30 minutes; keep the water bath stirring, slowly add the pentalhyde solution (concentration is 24.5 %, W W, W W / w) (The solid-liquid ratio of chitosan powder and pentaldehyde solution is 1g: 2.2ml), and the cross-linked response is 65 minutes; finally add N- (1,8-diemimimazazole and [1,2-a ] 喹 4 4-4-base) -1,2-ethyleine (the mass ratio of the chitosan powder is 1.87: 1), after the stirring reaction is 70min, add 5...

Embodiment 2

[0037] The difference between the preparation and embodiment 1 of a composite gel ball is:

[0038] CACO in Step S1 3 The mass ratio of the chitosan powder is 0.76: 1; the solid-liquid ratio of the chitosan powder and the pentalaldal solution is 1 g: 2ml; n- (1,8-two-gymiimazole and [1,2-a]基-4 -4-Bidth) -1,2-The mass ratio of ethyltramia and chitosan powder is 1.80: 1;

[0039] Step S2 The mass ratio of the middle product M and sodium alginate is 1: 0.8 to 1.2; the solid -liquid ratio of sodium alginate and calcium chloride solution is 0.1g: 18.4ml.

Embodiment 3

[0041] The difference between the preparation and embodiment 1 of a composite gel ball is:

[0042] CACO in Step S1 3 The mass ratio of the chitosan powder is 0.81: 1; the solid-liquid ratio of chitosan powder and pentalaldal solution is 1 g: 2.3ml; n- (1,8-diemimidazole and [1,2-a a ] 喹 4 啉 -4-base) -1,2-the mass ratio of ethyltramia and chitosan powder is 2.01: 1;

[0043] The quality ratio of the middle product M and sodium alginate in the step S2 is 1: 1.16; the solid -liquid ratio of sodium alginate and calcium chloride solution is 0.1g: 23.6ml.

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Abstract

The invention discloses a hydrogel-immobilized microbial preservation method, which relates to the technical field of immobilization, and comprises: immobilizing microorganisms in a hydrogel immobilized carrier through an embedding method, and storing them at normal temperature. Wherein, the carrier is a composite gel ball, including: embedding filler, including N‑(1,8‑dimethylimidazo[1,2‑a]quinoxaline‑4‑yl)‑1,2‑ethanedi Amine-modified chitosan microspheres; skeleton materials, including sodium alginate, form an interpenetrating or semi-interpenetrating network structure with embedding fillers. The microbial preservation method provided by the present invention uses hydrogel as a carrier, and is embedded in the carrier through immobilization technology, which can realize long-term storage and transportation at room temperature; and the prepared composite gel ball carrier has good compressive strength, High porosity, good stability, more conducive to microbial attachment.

Description

Technical field [0001] The present invention is a fixed biotechnology field, which involves a microbial preservation method with fixed hydrogel. Background technique [0002] A variety of bacteria is an important biological resource for the country, and it is also the basic material for production, teaching and scientific research.In recent years, more and more microbial bacteria have been studied and used in various fields such as food, medicine, and agriculture. The bacteria, as the source of production, are directly related to the production and quality of the product.In the field of microorganisms, whether it is basic scientific research work or biotechnology application research, the correct method and technique of storage of bacteria requires the quality and vitality of the bacteria.my country has started in the field of bacterial storage technology, and has not yet formed systemic operating procedures. There are mainly the following problems: the number of tests is limited...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/10C12N11/04C12N1/04C08B37/08B01J13/02
CPCC12N11/10C12N11/04C12N1/04C08B37/003B01J13/02
Inventor 陈庆国刘雨薇刘梅汪涛竺柏康
Owner ZHEJIANG OCEAN UNIV
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