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Low cost preparation method of deep sea elastase
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An elastase, deep-sea bacteria technology, applied in microorganism-based methods, biochemical equipment and methods, enzymes, etc., can solve the problems of high fermentation cost, affecting development and application, etc., and achieve the improvement of extracellular enzyme activity, good environmental benefits and Social benefits, the effect of reducing fermentation costs
Active Publication Date: 2014-07-23
SHANDONG UNIV
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Problems solved by technology
However, the fermentation cost of this enzyme using elastin is relatively high, which seriously affects the development and application of this enzyme.
Method used
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Embodiment 1
[0037] A low-cost preparation method of deep-sea bacterial elastase, the steps are as follows:
[0038] (1) Deep-sea bacteria (Pseudoalteromonas sp.) CF6-2 strain was activated and cultured in solid medium at 20°C for 1 day, then inoculated in liquid seed medium, and shaken at 15°C for 12 hours. Obtain seed culture fluid;
[0039] The components of the liquid seed culture medium are as follows, all in parts by weight:
[0041] (2) Inoculate the seed culture solution prepared in step (1) on the fermentation medium at a volume percentage of 1%, ferment and cultivate at a temperature of 18°C for 26 hours, and centrifuge at 10,000g for 10 minutes at 5°C to take the supernatant , to obtain a fermented liquid;
[0042] The fermentation medium components are as follows, all in parts by weight:
[0048] A low-cost preparation method of deep-sea bacterial elastase, the steps are as follows:
[0049] (1) Deep-sea bacteria (Pseudoalteromonas sp.) CF6-2 strain was activated and cultured in solid medium at a temperature of 15°C for 2 days, and shaken at 18°C for 15 hours to obtain a seed culture solution;
[0050] The components of the liquid seed culture medium are as follows, all in parts by weight:
[0051] 1.0 parts of peptone, 0.75 parts of yeast powder, 100 parts of artificial seawater, pH 8.0;
[0052] (2) Inoculate the seed culture solution prepared in step (1) on the fermentation medium at a volume percentage of 5%, ferment and cultivate at a temperature of 20°C for 30 hours, and centrifuge at 10,000g for 10 minutes at 5°C to take the supernatant , to obtain a fermented liquid;
[0053] The fermentation medium components are as follows, all in parts by weight:
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Abstract
The invention relates to a low cost preparation method of deep seaelastase, and the steps are as following: (1) inoculating deep seabacteria into a liquid seed medium for culture to obtain a seed nutrient solution; inoculating the seed nutrient solution into a fermentation medium for fermentation culture, then taking the supernatant to make a broth; (3) performing overnight dialysis, centrifugation and DEAE-SepharoseFast Flowchromatography and then gradient washing out to the broth, and taking the eluate of the first eluting peak to prepare the deep seabacteriaelastase. In the invention, elastinlaminin is replaced by scalded sorta as a byproduct of a slaughterhouse and is used for enzyme production fermentation of bacterial strain CF6-2 extracellularelastase, therefore the fermentation cost is reduced by more than 70%, and the method has good environment benefit and social benefit.
Description
technical field [0001] The invention relates to a low-cost preparation method of deep-sea elastase, in particular to a method for preparing deep-sea bacterial elastase by bovine heart tube fermentation, and belongs to the technical field of biotechnology. Background technique [0002] Elastase is a broad-spectrum proteolytic enzyme characterized by decomposing insoluble elastin. Since elastin is mainly distributed in connective tissues such as blood vessels, lungs and skin, elastase can be used as a treatment for hyperlipidemia, prevention and treatment. Biochemical drugs for atherosclerosis, elimination of skineschar, debris, etc., accurate treatment, safe and reliable; also can be used in food industry and cosmetic industry. Due to the similarity of elastin and gram-positive bacterial peptidoglycanpeptide partial sequence, elastase can sometimes be used as lysozyme with lytic activity. Lysozyme can be used as a natural foodpreservative. It has been widely used in aquat...
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