Hydrogel immobilized microorganism preservation method

A technology of microorganisms and hydrogels, which is applied in the direction of preservation of microorganisms, immobilization on/in organic carriers, and preparation of microspheres. Widely used in production and other issues to achieve the effect of maintaining microbial activity, easy operation, and preventing cell damage

Active Publication Date: 2021-07-27
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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  • Hydrogel immobilized microorganism preservation method
  • Hydrogel immobilized microorganism preservation method
  • Hydrogel immobilized microorganism preservation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] A kind of preparation of composite gel ball:

[0034] S1: Add chitosan powder to aqueous acetic acid solution (2.1%, w / w), stir until completely dissolved; add CaCO under magnetic stirring 3 (mass ratio of chitosan powder to 0.79:1), stirring for 12min, then ultrasonic for 15min; then add liquid paraffin (solid-liquid ratio of chitosan powder and paraffin is 1g:156mL), vigorously stir for 5min; 43 ℃ water bath Under the condition, add span-80 dropwise (the solid-liquid ratio of chitosan powder and span-80 is 1g: 2.1mL), stir 30min; / w) (the solid-to-liquid ratio of chitosan powder and glutaraldehyde solution is 1g: 2.2mL), carry out the crosslinking reaction for 65min; finally add N-(1,8-dimethylimidazo[1,2-a ]quinoxalin-4-yl)-1,2-ethylenediamine (mass ratio to chitosan powder is 1.87:1), after stirring for 70min, add 5M NaOH solution to adjust pH to 10.2, and adjust reaction temperature to Continue to stir and react at 74°C for 110 minutes; the reaction product is ri...

Embodiment 2

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

[0038] CaCO in step S1 3 The mass ratio of chitosan powder to chitosan powder is 0.76:1; the solid-liquid ratio of chitosan powder to glutaraldehyde solution is 1g:2mL; N-(1,8-dimethylimidazo[1,2-a] The mass ratio of quinoxalin-4-yl)-1,2-ethylenediamine to chitosan powder is 1.80:1;

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

Embodiment 3

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

[0042] CaCO in step S1 3 The mass ratio of chitosan powder to chitosan powder is 0.81:1; the solid-liquid ratio of chitosan powder to glutaraldehyde solution is 1g:2.3mL; N-(1,8-dimethylimidazo[1,2-a The mass ratio of ]quinoxalin-4-yl)-1,2-ethylenediamine to chitosan powder is 2.01:1;

[0043] Step S2 The mass ratio of intermediate product M to sodium alginate is 1:1.16; the solid-to-liquid ratio of sodium alginate to calcium chloride solution is 0.1 g:23.6 mL.

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Abstract

The invention discloses a hydrogel immobilized microorganism preservation method, and relates to the technical field of immobilized biologication. The hydrogel immobilized microorganism preservation method comprises the steps that microorganisms are immobilized in a hydrogel immobilized carrier through an embedding method, and storage and preservation are conducted at the normal temperature. The carrier is a composite gel ball, and comprises: an embedding filler, which comprises N-(1, 8-dimethylimidazo [1, 2-a] quinoxaline-4-yl)-1, 2-ethidene diamine modified chitosan microspheres, and a framework material which comprises sodium alginate, and the framework material and the embedding filler form an interpenetrating or semi-interpenetrating network structure. According to the microorganism preservation method provided by the invention, the hydrogel is used as a carrier and is embedded in the carrier through an immobilization technology, so that normal-temperature long-time storage, preservation and transportation can be realized; and the prepared composite gel ball carrier has good compressive strength, high porosity and good stability, and is more beneficial to microbial attachment.

Description

technical field [0001] The invention belongs to the technical field of immobilized biology, and in particular relates to a method for preserving microorganisms immobilized by hydrogel. Background technique [0002] Bacterial strains are important biological resources of the country and basic materials for production, teaching and scientific research. In recent years, more and more microbial strains have been studied and applied to various fields such as food, medicine, and agriculture. As the source of production, the quality of strains is directly related to the output and quality of products. In the field of microbiology, whether it is basic scientific research or applied research on biotechnology, correct strain preservation methods and technologies are required to ensure the quality and vitality of strains. Our country started relatively late in the field of strain preservation technology research, and has not yet formed a systematic operating procedure. The main proble...

Claims

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Application Information

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Patent Type & Authority Applications(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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