Isolation and purification method for high-yield of low-temperature catalase by antarctic marine bacillus n2a

A marine bacillus and catalase technology, applied in the field of marine microbial enzymes, can solve the problems of insufficiency, long steps, poor effect, etc., and achieve the effect of simple process, simple purification, fast effect and good effect

Inactive Publication Date: 2008-07-30
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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Problems solved by technology

Yumoto et al. first reported the properties of the low-temperature catalase (VRC) of the facultative psychrophilic bacteria Vibrio rumoiensis, and Lorentzen et al. studied the low-temperature catalase (VSC) from the fish pathogen Vibrio salmonicida, which made people preliminary Knowing low-temperature catalase, there are many differences between these two low-temperature enzymes and common low-temperature enzymes: the activi

Method used

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  • Isolation and purification method for high-yield of low-temperature catalase by antarctic marine bacillus n2a

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Embodiment 1

[0028] Strains and Culture Conditions

[0029] Strain n2a was isolated from Antarctic seawater and identified as a new species of Bacillus by 16SrRNA gene sequencing (GenBank accession number DQ508485). Growth and fermentation medium: Each 1L distilled water contains 8g peptone, 5gNaCl and 3g yeast extract, pH7.2. 11 shake flasks contain 200mL medium, 20°C, 200rpm shaker for 24h to late exponential growth phase. The cells were collected by centrifugation at 5000g at 4°C.

[0030] purification process

[0031]23g of bacteria were resuspended in 365ml pH7.5 50mmol / LTris.Cl (buffer A), and ultrasonically disrupted 10 times with 75W power (Vibra cell VCF1500, Sonics), each time for 1min with an interval of 2min. Centrifuge at 4°C to get the supernatant, add ammonium sulfate powder to 40% saturation, centrifuge to collect the supernatant, add ammonium sulfate to 70% saturation, and centrifuge at 4°C to collect the precipitate. Dialyze the precipitate against buffer A overnight, ...

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Abstract

The invention relates to a separation and purification method of high yield and low-temperature catalase of Antarctic marine Bacillus sp n2a. The invention can achieve electrophoretic pure low-temperature catalase through the processing of strains shaking culture, the ultrasonication ammonium sulfate precipitation, anion-exchange chromatography and gel filtration. With simple processing art, and fast and good effect, the invention can be used for separating and purifying low-temperature catalase in effective, reliable and economical way.

Description

technical field [0001] The invention relates to the field of marine microbial enzymes, in particular to a method for separating and purifying high-yield low-temperature catalase from Antarctic marine bacillus n2a. Background technique [0002] There are many kinds of low-temperature microorganisms living in the low-temperature environments of high mountains, deep seas and poles on the earth. They have evolved a complete set of adaptation mechanisms for survival and reproduction at around 0°C. Low temperature greatly reduces the rate of chemical reactions. How do low-temperature microorganisms maintain the normal progress of important biochemical reactions? , Enzymes that maintain normal activity at low temperatures are the key to ensuring the normal metabolism of low-temperature microorganisms. Low-temperature enzymes that catalyze important reactions are an important part of the study on the mechanism of low-temperature microorganisms adapting to low temperature. Generally...

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Application Information

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IPC IPC(8): C12N9/08C12N1/20C12R1/07
Inventor 孙谧王跃军郑媛朱重昊
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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