A kind of candida protease preparation and preparation method thereof

A technology of protease preparation and Candida, which is applied in the field of bioengineering, can solve the problems of low protease production, low protease activity, and few types of production strains, and achieve low operating costs, low impurity content, and large environmental advantages. Effect

Inactive Publication Date: 2018-08-14
SHIHEZI DEV ZONE SHIDA HEXU TECH DEV CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Though protease kind is many in the market at present, the kind of production bacterial strain used is less, be limited to the microorganism that food processing is used traditionally and confirm harmless bacterial strain; And the enzyme activity of the protease produced is not high

Method used

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  • A kind of candida protease preparation and preparation method thereof

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preparation example Construction

[0040] Specifically, the preparation method of a kind of Candida protease preparation that the embodiment of the present invention proposes, comprises the following steps:

[0041] 1) Making Candida astratus fermentation liquid from Candida astratus. This step specifically includes:

[0042] 11) Perform strain mutagenesis treatment on Candida astratus to obtain a mutagenized strain.

[0043] Specifically, pick a single colony of Candida stellata (Candidastellata) from the solid plate for cultivating Candida thallus, and inoculate the single colony of the selected Candida star on the yeast extract powder peptone glucose medium (Specifically, inoculate the single bacterium colony of Candida star in a 250-milliliter triangular flask containing 20-30 milliliters of yeast extract powder peptone glucose medium), at a temperature of 25-37 ° C at 100-300 rpm Cultivate on a shaker / min for 24-48 hours to obtain a fermentation broth. Place the fermentation broth in a centrifuge, centr...

Embodiment 1

[0075] The present embodiment prepares a kind of Candida protein preparation, and concrete steps are as follows:

[0076] 1) strain mutagenesis

[0077] 11) Using ultraviolet light to mutagenize Candida: pick a single colony of Candida stellata from the solid plate of Candida thallus, inoculate it in a culture medium containing 20 ml of yeast extract powder peptone glucose In a 250-ml Erlenmeyer flask with base, cultivate it on a shaking table at 200 rpm at a temperature of 30°C for 36 hours to obtain a fermentation broth; place the fermentation broth in a centrifuge and centrifuge at a speed of 5000 rpm for 6 minutes Finally, discard the supernatant, and resuspend the cells in the lower layer with sterile water to obtain the bacterial liquid; make the cell concentration in the bacterial liquid reach 2×10 8 cells / ml, draw 3 ml of bacterial solution and place it in a 4°C refrigerator as a control; draw 5 ml of the remaining bacterial solution and add it to a sterile plate, and...

Embodiment 2

[0094] The present embodiment prepares a kind of Candida protein preparation, and concrete steps are as follows:

[0095] 1) strain mutagenesis

[0096] 11) Ultraviolet mutagenesis of Candida: Pick a single colony of Candida stellata from the solid plate of Candida thallus, inoculate it in 25 ml of yeast extract powder peptone glucose medium In a 250-ml Erlenmeyer flask, cultivate it on a shaker at 100 rpm at a temperature of 35°C for 40 hours to obtain a fermentation broth; place the fermentation broth in a centrifuge and centrifuge at a speed of 4000 rpm for 8 minutes Discard the supernatant, and resuspend the lower layer of cells with sterile water to obtain the bacterial liquid; make the cell concentration of the bacterial liquid reach 2×10 8 cells / ml, draw 3 ml and place it in a 4°C refrigerator as a control; draw 5 ml of the remaining bacterial solution and add it to a sterile plate, and divide 2×10 5 J / mm 2 The metering treatment of the aseptic plate, the aseptic pla...

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Abstract

The invention discloses a candida stellatta protease preparation and a preparation method thereof, develops a novel strain for preparing the candida stellatta protease preparation, and belongs to the field of bio-engineering. According to the main technical scheme of the invention, the preparation method of the candida stellatta protease preparation comprises the following steps of: preparing a candida stellatta fermentation liquor by using candida stellatta, and adding a salt into the candida stellatta fermentation liquor to separate out candida stellatta protease; dissolving the candida stellatta protease in a buffer solution, dialyzing, and purifying to obtain a candida stellatta protease liquor; ultra-filtrating and concentrating the candida stellatta protease liquor to obtain a candida stellatta protease concentrate; adding an inorganic salt and an auxiliary material (carrier) into the candida stellatta protease concentrate, and mixing fully to obtain a mixture; and freezing and drying the mixture to obtain the candida stellatta protease preparation. The preparation method provided by the invention is low in cost, is environment-friendly, is suitable for large-scale industrial production of acid protease preparations, and the candida stellatta protease preparation prepared by adopting the preparation method has high activity and good stability.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a method for producing a candida protease preparation by using a microbial liquid fermentation method. Background technique [0002] Protease is a kind of enzyme that catalyzes the hydrolysis of protein. Protease is widely used in detergent, feed, food industry, wine brewing, leather industry and pharmaceutical fields; in addition, protease is also one of the most widely used enzyme preparations. Used in the production of detergents, tanning, fur, protein hydrolyzate, brewing, soy sauce, textiles, pharmaceuticals, cosmetics, etc. and the extraction of biological materials. The wide application of protease not only simplifies the production process of the industry, saves costs, improves product yield, but also makes positive contributions to improving the environment and reducing carbon dioxide emissions. In recent years, with the enhancement of people's awareness of envir...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/60C12R1/72
CPCC12N9/60
Inventor 史学伟徐亚男王军程卫东
Owner SHIHEZI DEV ZONE SHIDA HEXU TECH DEV CO LTD
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