Bacterial strain for producing acid protease through liquid fermentation and application of bacterial strain

An acid protease and liquid fermentation technology, which is applied in the field of Aspergillus niger strain and its liquid fermentation production of acid protease, and can solve the problems of limiting the wide application of acid protease, high overall cost, limited production output and the like

Active Publication Date: 2021-06-11
安琪酶制剂(宜昌)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The production output is also limited by the solid-state fermentation production site, coupled with insufficient product purity, low yield in the extraction step, and high overall cost, which limits the wide application of acid protease to a certain extent

Method used

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  • Bacterial strain for producing acid protease through liquid fermentation and application of bacterial strain
  • Bacterial strain for producing acid protease through liquid fermentation and application of bacterial strain
  • Bacterial strain for producing acid protease through liquid fermentation and application of bacterial strain

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Experimental program
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Effect test

Embodiment 1

[0062] The acquisition and mutagenesis of embodiment 1 Aspergillus niger ESP1023

[0063] A Aspergillus niger strain isolated from the soil around the fermented bean curd factory in Yichang City, Hubei Province was mutated and domesticated by combining ultraviolet mutagenesis technology and ARTP mutagenesis technology to obtain the new acid-producing protease ability provided by the present invention. Excellent bacterial strains, the specific method is as follows.

[0064] 1. The Aspergillus niger strain isolated from soil is subjected to ultraviolet mutagenesis:

[0065] Bacterial suspension preparation: Wash out the spores of eggplant bottles cultivated to the logarithmic phase with 50mL of normal saline; shake evenly; take 15ml of spore suspension and pour it into a sterile plate;

[0066] In an environment without visible light, place the plate containing the spore liquid vertically under a UV lamp with a distance of 28 cm, an irradiation time of 270 s, and a power of 40 ...

Embodiment 2

[0088] Embodiment 2 Aspergillus niger ESP1023 bacterial strain liquid fermentation produces acid protease

[0089] Strain activation: inoculate the glycerol tube seeds of the Aspergillus niger ESP1023 strain into fresh PDA eggplant bottle medium, and culture at 29°C for 6 days.

[0090] Seed shake flask fermentation: Pick a large ring of mature seed lawn from the eggplant bottle and put it into freshly prepared seed shake flask culture medium. Under the condition of 29°C, 180rpm shake flask culture for 30h, the weight volume ratio of seed shake flask medium (g / ml), the components are as follows: glucose 1.5%, corn steep liquor 2.0%, ammonium sulfate 0.5%, magnesium sulfate 0.1%, potassium dihydrogen phosphate 1.5%, calcium chloride 0.05%, trace element A mother liquor 0.04%; The pH is controlled at 5.5, sterilized at 115°C for 30 minutes, and the trace element A mother solution is made of the following components: in terms of mass volume ratio (g / ml), 0.6% iron citrate, 0.08% ...

Embodiment 3

[0096] Embodiment 3 Aspergillus niger ESP1023 bacterial strain liquid fermentation produces acid protease

[0097] Strain activation: inoculate the glycerol tube seeds of the Aspergillus niger ESP1023 strain into fresh PDA eggplant bottle medium, and culture at 29° C. for 5 days.

[0098] Seed shake flask fermentation: Pick a large ring of mature seed lawn from the eggplant bottle and put it into freshly prepared seed shake flask culture medium. Under the condition of 29°C, 180rpm shake flask culture for 30h, the weight volume ratio of seed shake flask medium (g / ml), the components are as follows: glucose 2.0%, corn steep liquor 2.0%, ammonium sulfate 0.8%, magnesium sulfate 0.1%, potassium dihydrogen phosphate 0.95%, calcium chloride 0.05%, trace element A mother liquor 0.04%; The pH is controlled at 5.0, sterilized at 115°C for 30 minutes, and the trace element A mother solution is made of the following components: in terms of mass volume ratio (g / ml), 0.6% iron citrate, 0.0...

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Abstract

The invention provides an aspergillus niger strain and an application thereof in fermentation production of acid protease. The invention provides an Aspergillus niger ESP1023 strain (Aspergillus niger ESP1023), which is characterized in that the Aspergillus niger ESP1023 strain (Aspergillus niger ESP1023) is preserved in the China Center for Type Culture Collection (CCTCC), and the preservation number is CCTCC NO: M 2018829. By utilizing the Aspergillus niger ESP1023 strain and the method for producing the acid protease through liquid fermentation, the zymoprotein purity in the obtained fermentation liquor is high, and the enzyme activity is high.

Description

technical field [0001] The invention relates to the technical field of microbial fermentation, in particular to an Aspergillus niger strain and a method for producing acid protease by liquid fermentation thereof. Background technique [0002] Acid proteinase is a class of aspartic acid protease with an optimum pH of about 2.5-5.5, which was first discovered by Yoshida in Aspergillus niger in 1945. Acid protease widely exists in molds and basidiomycetes, and its relative molecular weight is about 30KDa-40KDa. The source of acid protease is mainly the secretion of animal viscera and specific microorganisms, including pepsin, rennet and some microbial proteases. [0003] There are many types of acid protease-producing strains. At present, the strains used for the production of acid protease mainly include Aspergillus niger, Aspergillus oryzae, Saccharomyces cerevisiae, Bacillus subtilis, etc., among which Aspergillus niger is the main one. The produced acid proteases all hav...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N9/62C12R1/685
CPCC12N1/14C12N9/62
Inventor 李汉文熊涛李知洪喻晨吴尧胡悦栾春艳余华顺姚鹃
Owner 安琪酶制剂(宜昌)有限公司
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