A preparation method of glycerol kinase

A technology of glycerol kinase and glycerol, which is applied in the direction of biochemical equipment and methods, enzymes, transferases, etc., can solve the problems of unstable enzyme production characteristics and low activity, achieve rapid preparation, improve preparation efficiency, and shorten the preparation process.

Inactive Publication Date: 2019-01-15
WUHAN HANHAI NEW ENZYMES BIOLOGICAL TECH CO LTD
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the deficiencies of the prior art, the present invention provides a method for preparing glycerol kinase, which solves the problems of unstable enzyme production characteristics and low activity in existing natural bacteria

Method used

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Embodiment Construction

[0017] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some of the embodiments of the present invention, but not all of them. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

[0018] The embodiment of the present invention provides a technical solution: a preparation method of glycerol kinase, which specifically includes the following steps:

[0019] Next steps:

[0020] S1. Fermentation tank culture: first inoculate the glycerol-preserved bacteria in the LB medium of the test tube, and culture it on a shaker at 37°C and 200r / min for 10h-12h;

[0021] S2, re-inoculate in the shake flask fermentation medium according to 2% inoculum amount, and cultivate for 10h-12h under the same conditions as the seed st...

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Abstract

The invention discloses a preparation method of glycerol kinase, in particular comprising the following steps: S1, culturing in a fermentor: firstly, inoculating glycerol-preserving bacteria in a testtube LB culture medium, culturing in a shaking table at 37 DEG C and 200r / min for 10h; and secondly, culturing the glycerol-preserving bacteria in a shaking table at 37 DEG C and 200r / min for 10h. 12h; S2, inoculated in shake flask fermentation medium according to 2% inoculum, cultured in the same condition for 10h-12 hour as seed strain; S3, fermentor cultured until glucose is consumed completely, adding 6.0 g / L lactose to induce fermentation. The invention relates to the technical field of medical reagents. The preparation method of glycerol kinase can make the enzyme-producing characteristics of natural bacteria more stable and greatly enhance the activity of glycerol kinase, and realizes the expression of the fermentation and culture conditions of recombinant glycerol kinase r-GK engineering strain, the establishment of purification process and the quality control detection of the enzyme, these research results will be a solid foundation for the development and utilization of theenzyme.

Description

technical field [0001] The invention relates to the technical field of medical reagents, in particular to a preparation method of glycerol kinase. Background technique [0002] The content of triglycerides in serum is an important indicator for clinical diagnosis of cardiovascular system diseases. In view of the many shortcomings of chemical method determination, enzyme-linked kits with high specificity, high sensitivity, rapidity, automation and other advantages have been successively used. Instead, glycerol kinase (E.C 2.7.1.30) is one of the key enzymes in the enzyme-linked kit for determining triglycerides, but the enzyme-linked kit requires high purity for the enzymes involved in the reaction. Interfering enzymes such as NADH oxidase and catalase are also required to be controlled within a very low range, otherwise it will affect the assay results of the kit. We have studied glycerol kinase, and the purpose is to put this enzyme into Practical application, but due to t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/12
CPCC12N9/1205C12Y207/0103
Inventor 谢渊罗漫杰杨广宇
Owner WUHAN HANHAI NEW ENZYMES BIOLOGICAL TECH CO LTD
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