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Separating and purifying method for low-temperature alkaline lipase

A technology for separation and purification of lipase, which is applied in the direction of hydrolytic enzymes, etc., can solve the problems of separation and purification of lipase, limit the development of marine lipase, etc., and achieve the effect of high purity and simplified purification process

Inactive Publication Date: 2005-05-11
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the further development of marine lipase in the preparation and application is limited due to the problems of seawater culture and lipase separation and purification in marine microorganisms.

Method used

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  • Separating and purifying method for low-temperature alkaline lipase
  • Separating and purifying method for low-temperature alkaline lipase

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Embodiment 1

[0039] The fermentation medium is 4% (Wt) of bean cake powder, 4% (Wt) of rice bran, 4% (Wt) of peanut oil, 0.5% (Wt) of crude peanut oil, MgSO 4 0.05% (Wt), KH2PO 4 0.2% (Wt), PH5.0, culture temperature 26 ± 1 ℃, culture time 20-26 hours, inoculate the cold-adapted marine yeast fungus BOhai-Sea-9145 shock culture of the present invention, then high-yield in the culture medium Centrifuge the culture solution of marine microbial low-temperature alkaline lipase at 4°C at 1000 rpm / min to obtain a supernatant containing marine microbial low-temperature alkaline lipase of the present invention, and the specific activity of the lipase in the supernatant is about 12652U / ml.

[0040]Add chloroform to the above supernatant with a volume ratio of chloroform to supernatant of 5:100 for thorough stirring, let stand at 4° C. for 30 minutes, and centrifuge at 4000 rpm for 40 minutes to remove impurities and collect the supernatant. The obtained supernatant was ultrafiltered through a 10...

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Abstract

The invention was involved in the new purification method of low-temperature alkalinous lipase. The purification was performed with a four-step procedure consisting of extracting the fermentation upper liquid with methenyl chloride, ultrafiltrating with 100kda hollow fibre column, contacting with 10kda hollow fibre column and using CM Sephanose FF positive ion exchange chromatography and finally achieved low-temperature alkalinous lipase. The method got rid of the influence of inducer to enzyme purifying, simplify purification method with high purity and can be used as the new industrialized purification method.

Description

technical field [0001] The present invention relates to a method for separation and purification of marine microorganisms, in particular to a method for separation and purification of novel low-temperature alkaline lipase Background technique [0002] In recent years, with the development of biotechnology, searching for enzymes with new properties has become a hot spot in international enzyme research. Because marine organisms grow in the unique environment of the ocean, especially marine organisms and microalgae living in extreme environments, they can produce abundant extreme enzymes. Compared with enzymes produced by terrestrial microorganisms, marine enzymes have more unique enzymatic properties, so they have attracted great attention from researchers at home and abroad. Lipase (glyceride hydrolase, EC3.1.3) is a special class of ester bond hydrolase, which can catalyze the hydrolysis of natural substrate oil (triglyceride) to produce fatty acids and glycerol. Therefor...

Claims

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Application Information

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IPC IPC(8): C12N9/20
Inventor 王跃军孙谧王海英阎晓玲
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI