Method for preparing phosphoglyceric oxidase using inducing culture substrate chain coccus

A technology of phosphoglycerol oxidase and inducing medium, which is applied in the field of producing phosphoglycerol oxidase by using inducing medium

Inactive Publication Date: 2007-11-14
王腾 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 1997, Feng Yongmei, Institute of Microbiology, Chinese Academy of Sciences, etc. reported the research on the strain se

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] One, make culture medium (weight percentage)

[0018] Slant induction medium (%): yeast extract 2.5, peptone 2.0, glucose 0.5, dipotassium hydrogen phosphate 0.8, glycerol 0.5, agar 3.0, magnesium sulfate 4, manganese chloride 1, sodium chloride 20, pH 6.8 0.4 ml of saline solution;

[0019] Seed medium (%): yeast extract 2.5, peptone 2.0, glucose 0.8, dipotassium hydrogen phosphate 0.8, glycerol 0.5, magnesium sulfate 4, manganese chloride 1, sodium chloride 20, pH 6.8 mixed salt solution 0.4 ml ;

[0020] Fermentation medium (%): yeast extract 2.5, peptone 2.0, dipotassium hydrogen phosphate 2.0, glycerol 0.5, magnesium sulfate 4, manganese chloride 1, sodium chloride 20, 0.4 ml of mixed salt solution of pH 6.8;

[0021] 2. Cultivate strains

[0022] After the strain was cultured on a slant medium at 32°C for 28 hours, it was inserted into the seed medium, and cultured with shaking at 32°C and 200 rpm for 24 hours. The seed solution was added to the fermentation m...

Embodiment 2

[0030] 1. Production medium (%):

[0031] Slant induction medium: yeast extract 0.5, peptone 1.0, glucose 0.05, dipotassium hydrogen phosphate 0.2, glycerin 0.1, agar 1.5, magnesium sulfate 4, manganese chloride 1, sodium chloride 20, pH 6.8

[0032] Mixed salt solution 0.1 ml;

[0033] Seed medium: yeast extract 0.5, peptone 1.0, glucose 0.1, dipotassium hydrogen phosphate 0.2, glycerin 0.1, magnesium sulfate 4, manganese chloride 1, sodium chloride 20, 0.1 ml of mixed salt solution with pH 6.8;

[0034] Fermentation medium: 0.5 yeast extract, 1.0 peptone, 0.5 dipotassium hydrogen phosphate, 0.1 glycerol, 4 magnesium sulfate, 1 manganese chloride, 20 sodium chloride, 0.1 ml of mixed salt solution with pH 6.8;

[0035] 2. Cultivate strains

[0036] After the strain was cultured on the slant medium at 25°C for 18 hours, it was inserted into the seed medium, and cultured with shaking at 25°C and 200 rpm for 18 hours. The seed solution was added to the fermentation medium at a...

Embodiment 3

[0039] 1. Production medium (%)

[0040] Slant induction medium: yeast extract 1.0, peptone 1.5, glucose 0.25, dipotassium hydrogen phosphate 0.5, glycerin 0.12, agar 2.0, magnesium sulfate 4, manganese chloride 1, sodium chloride 20, pH 6.8 mixed salt solution 0.2 ml;

[0041] Seed medium: 0.5 yeast extract, 1.0 peptone, 0.5 glucose, 0.5 dipotassium hydrogen phosphate, 0.1 glycerol, 4 magnesium sulfate, 1 manganese chloride, 20 sodium chloride, 0.2 ml of mixed salt solution with pH 6.8;

[0042] Fermentation medium: yeast extract 0.5, peptone 1.0, dipotassium hydrogen phosphate 2.0, glycerin 0.1, magnesium sulfate 4, manganese chloride 1, sodium chloride 20, 0.2 ml of mixed salt solution with pH 6.8;

[0043] 2. Cultivate strains

[0044] After the strain was cultured on the slant medium at 30°C for 22 hours, it was inserted into the seed medium, and cultured at 30°C at 200 rpm for 20 hours. The seed solution was added to the fermentation medium at a ratio of 10%, and cultur...

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Abstract

This invention is with the guide hammer bacterium of culture medium preparation Lin the method of sour glycerine oxidize enzyme. The group of culture medium divides and cultivates condition: Bevel guides culture medium, draw thing by yeast , egg white Dong , glucose , Lin the mixture salt solution of 2 sour hydrogen potassiums, glycerines, agars, the magnesiums of sulphuric acid, chlorination manganese, common salts and pH6.8s forms; Seed culture medium draws thing and egg white by yeast Dong , glucose , Lin the mixture salt solution of 2 sour hydrogen potassiums, glycerines, the magnesiums of sulphuric acid, chlorination manganeses, common salts and pH6.8s forms; Seed culture medium draws thing and egg white by yeast Dong , glucose , Lin the mixture salt solution of 2 sour hydrogen potassiums, glycerines, the magnesiums of sulphuric acid, chlorination manganeses, common salts and pH6.8s forms; Ferment culture medium draws thing and egg white by yeast Dong , Lin the mixture salt solution of 2 sour hydrogen potassiums, glycerines, the magnesiums of sulphuric acid, chlorination manganeses, common salts and pH6.8s forms. Bacterial strain is cultivated by bevel, seed is cultivated , revolving shaking table ferments to train , enlarging again collect bacterium body after training , adoption assembles glycol - the two water of ammonium sulphate appearance extraction and the layer of two steps Xi purification gets Lin sour glycerine oxidize enzyme.

Description

technical field [0001] The invention relates to a method for producing oxidase, in particular to a method for producing phosphoglycerol oxidase with an induction medium. Background technique [0002] The content of triglycerides in serum has been recognized as an important index in the clinical diagnosis of hyperlipidemia and heart disease. The glycerol phosphate oxidase method for the determination of triglyceride content has the advantages of high specificity, small amount, simplicity, rapidity, and automatic instrument determination. It was once recommended by the Chinese Medical Association as a routine method for the determination of serum triglycerides in clinical laboratories. [1，2] . Glycerol phosphate oxidase (L-a-Glycerophosphate Oxidase, GPO, EC 1.1.3.21), also known as glycerol phosphate oxidase, is the last enzyme in the determination reaction of the triglyceride kit, which requires higher specificity and enzyme activity and specific activity. At present, the...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/02C12R1/46
Inventor 王腾孟延发
Owner 王腾
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