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Optimization method of culture conditions of alkaline pectinase gene engineering bacteria

A technology of genetically engineered bacteria and culture conditions, applied in the field of optimization of the culture conditions of alkaline pectinase genetically engineered bacteria, can solve problems such as increasing production costs, achieve great theoretical and application value, and improve market competitiveness.

Inactive Publication Date: 2010-07-21
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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Problems solved by technology

More and more research units have carried out effective attempts on the molecular biology operation of alkaline pectinase under the general trend of genetically engineered bacteria being applied to industrial production. Most of the molecular biology operations mainly focus on the expression system of Escherichia coli, or The Bacillus subtilis expression system is used as the research object, but most of the selection of vectors and hosts show some shortcomings when the purpose of production is for example. increase production cost

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  • Optimization method of culture conditions of alkaline pectinase gene engineering bacteria
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  • Optimization method of culture conditions of alkaline pectinase gene engineering bacteria

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Embodiment Construction

[0031] Below in conjunction with the examples, the present invention is further described, the following examples are illustrative, not limiting, and the protection scope of the present invention cannot be limited by the following examples.

[0032] 1. Experiment description

[0033] 1. The experimental strain is genetically engineered strain WB600 / pWB-pelG 2 521, the strain is Bacillus subtilis (with six protease deficient types), the strain and construction method have been applied for a patent, the patent application number is: 200910068617.6, and the application date is: April 24, 2009. The production strain was cultured at a constant temperature of 37°C for 24 hours on an agar slant. The bacterium had grown completely on the slant and was milky white in appearance.

[0034] 2. Experimental strain WB600 / pWB-pelG 2521 slant medium: peptone 1%, yeast extract 0.5%, NaCl 1%, pH value 6.8-7.0, agar powder 1.5%.

[0035] 3. Experimental strain WB600 / pWB-pelG 2 521 seed mediu...

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Abstract

The invention relates to an optimization method of culture conditions of alkaline pectinase gene engineering bacteria, comprising the following steps: (1) adopting a single-factor experiment to optimize the fermentation medium formation of the alkaline pectinase gene engineering bacteria to obtain a preliminarily optimized fermentation medium; (2) optimizing a target strain medium with a response surface methodology to obtain the target strain medium; and (3) optimizing the fermentation culture conditions to the target strain by a post-optimizing medium to obtain the optimization culture conditions. The invention gropes a set of optimal fermentation culture conditions to ensure that enzymatic activity produced by engineering bacteria fermentation reaches 758.7825U / ml which is improved by 50 times compared with the activity of alkaline pectinase produced by original wide strains and is improved by 3 times compared with the enzymatic activity of alkaline pectinase produced by LB fermentation medium fermentation gene engineering bacteria, so that the invention has an important meaning for improving the market competitiveness of industrial enzyme.

Description

technical field [0001] The invention belongs to the field of microbial engineering, in particular to a method for optimizing the culture conditions of alkaline pectinase genetically engineered bacteria. Background technique [0002] Pectinase is a general term for a class of enzymes that decompose pectin, and is a compound enzyme containing multiple components. Pectinase is widely distributed in higher plants and microorganisms, but the extraction from plants has been limited by resources and technical level and has not been truly industrialized. Therefore, pectinase from microorganisms has become a research hotspot. The research on acid pectinase at home and abroad has a certain basis, but there are few research reports on alkaline pectinase, especially pectate lyase. [0003] Alkaline pectinase refers to the pectinase with the most suitable pH in the alkaline range. It is an emerging content in the field of pectinase research. It is mainly used in the textile industry as ...

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Application Information

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IPC IPC(8): C12N9/26C12R1/125
Inventor 路福平郝育杰刘逸寒王春霞肖静周浩蒋彦洁王珊
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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