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Deoxidizing method of fluid medium

A technology of liquid medium and volume ratio, which is applied in the field of anaerobic microorganism cultivation and fermentation test, and can solve the problems of slow progress in anaerobic microorganism technology and methods, backward understanding and utilization of anaerobic microorganisms, separation and pure breed of anaerobic microorganisms Problems such as difficulty in cultivating to achieve the effect of avoiding poisoning

Inactive Publication Date: 2018-04-13
ANHUI RUISIWEIER TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the difficulty in the isolation and pure culture of anaerobic microorganisms, and the slow progress in the technology and methods of studying anaerobic microorganisms, especially in some small laboratories, human beings' understanding and utilization of anaerobic microorganisms lag far behind that of human beings. Research work on aerobic and facultative anaerobic microorganisms

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The deoxygenation method of the liquid medium of the present embodiment is to carry out as follows:

[0021] S1, prepare the resazurin aqueous solution as an anaerobic indicator, and the anaerobic indicator is dark blue in an aerobic state, pink in a micro-aerobic state, and colorless in anaerobic state; The mass volume ratio of Azure and double distilled water is 1.00g / L;

[0022] S2. Add L-cysteine ​​hydrochloride, biological buffer MES and the anaerobic indicator obtained in S1 to an anaerobic bottle equipped with 1L liquid medium and mix well; wherein, L-cysteine The mass volume ratio of salt to liquid medium is 0.50g / L, the mass volume ratio of biological buffer MES to liquid medium is 0.50g / L, and the volume ratio of anaerobic indicator to liquid medium is 0.50mL / L ;

[0023] S3, feed nitrogen into the bottom of the anaerobic bottle of S2 for 50 minutes, pretreat part of the dissolved oxygen in the liquid medium, and change the color of the liquid medium from da...

Embodiment 2

[0027] The deoxygenation method of the liquid medium of the present embodiment is to carry out as follows:

[0028] S1, prepare the resazurin aqueous solution as an anaerobic indicator, and the anaerobic indicator is dark blue in an aerobic state, pink in a micro-aerobic state, and colorless in anaerobic state; The mass volume ratio of Azure and double distilled water is 1.50g / L;

[0029] S2. Add L-cysteine ​​hydrochloride, biological buffer MES and the anaerobic indicator obtained in S1 to an anaerobic bottle equipped with 1L liquid medium and mix well; wherein, L-cysteine The mass volume ratio of salt to liquid medium is 1.00g / L, the mass volume ratio of biological buffer MES to liquid medium is 1.00g / L, and the volume ratio of anaerobic indicator to liquid medium is 1.00mL / L ;

[0030] S3, feed nitrogen into the bottom of the anaerobic bottle of S2 for 60 minutes to pretreat part of the dissolved oxygen in the liquid medium, so that the color of the liquid medium changes ...

Embodiment 3

[0034] The deoxygenation method of the liquid medium of the present embodiment is to carry out as follows:

[0035] S1, prepare the resazurin aqueous solution as an anaerobic indicator, and the anaerobic indicator is dark blue in an aerobic state, pink in a micro-aerobic state, and colorless in anaerobic state; The mass volume ratio of Azure and double distilled water is 2.00g / L;

[0036] S2. Add L-cysteine ​​hydrochloride, biological buffer MES and the anaerobic indicator obtained in S1 to an anaerobic bottle equipped with 1L liquid medium and mix well; wherein, L-cysteine The mass volume ratio of salt to liquid medium is 1.50g / L, the mass volume ratio of biological buffer MES to liquid medium is 1.50g / L, and the volume ratio of anaerobic indicator to liquid medium is 1.50mL / L ;

[0037] S3, feed nitrogen into the bottom of the anaerobic bottle of S2 for 70 minutes, pretreat part of the dissolved oxygen in the liquid medium, and change the color of the liquid medium from da...

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Abstract

The invention provides a deoxidizing method of a fluid medium. The method comprises the following steps: S1, preparing a resazurin water solution to serve as an anaerobic indicator which takes on a dark blue color or a light blue color in an oxygen dissolution state, takes on a pink color in a slight oxygen dissolution state, and is colorless in an oxygen dissolution-free state; S2, adding L-cysteine hydrochloride, a biological buffering agent MES (morpholineethanesulfonic acid) and an anaerobic indicator into an anaerobic bottle with 1L of the fluid medium and mixing uniformly; S3, introducing nitrogen gas to the bottom of the anaerobic bottle in the S2 for 50-100min, pretreating a part of dissolved oxygen in the fluid medium to change the color of the fluid medium from the dark blue color to the light blue color or the light pink color; and S4, enabling the fluid medium of which a part of the dissolved oxygen is removed to stand, and consuming the rest of dissolved oxygen in the anaerobic bottle through the added L-cysteine hydrochloride in the S2 to change the color of the fluid medium from the light blue color or the light pink color to an achromatic color, thus completing deoxygenization. The deoxidizing method can be used for removing the dissolved oxygen in the fluid medium, and is conducive to enrichment and cultivation research of anaerobic micro-organisms.

Description

technical field [0001] The invention relates to the technical field of cultivation and fermentation test of anaerobic microorganisms, in particular to the enrichment and cultivation of strict anaerobic microorganisms, and more specifically to a method for deoxygenating a liquid medium. Background technique [0002] Oxygen toxicity mechanism of anaerobic microorganisms - SOD theory: strict anaerobic microorganisms are not killed by gaseous oxygen, but die because they cannot remove the toxicity of certain oxygen metabolites. During the reduction of oxygen to water, certain toxic intermediates can be formed, such as hydrogen peroxide (H 2 o 2 ), superoxide anion (O 2 °—) etc. Superoxide anion is active oxygen, which has the properties of both molecules and ions. It has a strong reaction force and is extremely unstable. It can destroy membranes and important biological macromolecules, and cause poisoning or death to microorganisms. Aerobic microorganisms have enzymes to deg...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/00
CPCC12N1/00
Inventor 王艳丽何宏魁李安军刘国英周庆伍王录刘开放李静心吴翠芳
Owner ANHUI RUISIWEIER TECH
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