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A halotolerant bacillus producing alkaline protease and its method and application for producing alkaline protease

A technology of bacillus halide and protease, applied in the field of microorganisms, can solve the problems of limited washing efficiency and reduced stability, etc.

Active Publication Date: 2022-03-18
SHANXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Subtilisins currently added to detergents include Savinase™ (Subtilisin 309), Subtilisin Novo (BPN′), Alcalase™ (Subtilisin Carlsberg), Maxacal™ (Novozymes A / S, Denmark), BLAP Sb (Henkel, Germany), and Properase™ (Genecor Int. USA), which are generally stable at elevated temperature and pH conditions, however, most of these proteases are less stable in the presence of liquid detergents and have limited wash efficiency

Method used

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  • A halotolerant bacillus producing alkaline protease and its method and application for producing alkaline protease
  • A halotolerant bacillus producing alkaline protease and its method and application for producing alkaline protease
  • A halotolerant bacillus producing alkaline protease and its method and application for producing alkaline protease

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Experimental program
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Effect test

Embodiment 1

[0025] Embodiment 1: strain screening

[0026] Bacteria were isolated and screened from farmland soil samples in Zuoquan County, Jinzhong City, Shanxi Province (37°07′N, 113°37′E).

[0027] Preliminary screening: Weigh 1g of soil sample and place it in an oven at 80°C for 2h, then use sterile water according to 10 -1 The gradient was serially diluted to 10 -6 , spread on a non-resistant milk plate, culture at 37°C for 12 hours, a clear transparent circle will be formed around the protease-producing colony, and the strain with the largest diameter ratio of the transparent circle to the colony will be selected as the strain for re-screening; primary screening and co-enrichment screening Up to 80 strains were named AS1-AS20, BS1-BS20, CS1-CS20, DS1-DS20 respectively. The milk medium is LB medium containing 2.0% (w / v) skimmed milk powder.

[0028] Re-screening: Inoculate the strains obtained from the primary screening in the fermentation medium for 12 hours at 37°C and centrifu...

Embodiment 2

[0033] Embodiment 2: identification of protease producing bacteria DS5

[0034] Observations on the DS5 morphological characteristics obtained above are as follows: figure 1 As shown, the colonies are milky white, opaque, rough and wrinkled, and round in shape. DS5 is Gram-positive, and the cells are short rod-shaped (0.5-1.0µm×3.0-6.0µm), arranged individually; the strain forms a round, milky white, opaque colony on the skimmed milk powder plate on the surface of LB medium Produces distinct transparent circles.

[0035] Physiological and biochemical identification results of the strain showed that it was Bacillus halotolerant ( Bacillus halotolerans ), the physiological and biochemical identification results are shown in Table 2.

[0036] Table 2: Halotolerant Bacillus ( Bacillus halotolerans ) Physiological and biochemical identification results

[0037]

[0038] The above physiological and biochemical characteristics detection methods are as follows:

[0039] a) G...

Embodiment 3

[0079] Example 3: Fermentation culture of protease-producing bacteria DS5: the fermented strains obtained from the screening were inoculated in the seed medium for cultivation, and then inoculated in the fermentation medium for fermentation. Specifically: inoculate the DS5 strain slant in the seed medium, cultivate at 37°C and 200rpm for 10 hours, then add the seed solution to the fermentation medium with 4% inoculation amount (10 g of dextrin and soluble starch per liter of liquid medium) 20g, yeast extract 10g, sodium chloride 5g, steam sterilization at 121°C for 20min), fermentation culture is 300mL of fermentation medium per 3L shake bottle, culture conditions: 37°C, 200rpm, fermentation culture for 12 hours.

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Abstract

The invention belongs to the technical field of microorganisms, and provides an alkaline protease-producing halal-tolerant Bacillus strain DS5 and a method and application thereof for producing alkaline protease. The alkaline protease-producing Bacillus halobacterium DS5 is Bacillus halotolerant ( Bacillus halotolerans ), deposited in the Guangdong Provincial Microbial Culture Collection and Management Center on August 17, 2020, with the deposit number: GDMCC No.61148; the deposit address is: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangdong City. The strains were isolated and screened from farmland soil samples in mountainous areas of Shanxi. The strain produces alkaline protease, which is resistant to salt and high temperature, and the protease produced is extracellular; Alkali resistance and high temperature resistance characteristics, can be used as a washing additive, with excellent washing effect.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to an alkaline protease-producing halotolerant bacillus and a method and application thereof for producing alkaline protease. The alkaline protease-producing halotolerant bacillus is strain DS5. Background technique [0002] Protease is a kind of enzyme that catalyzes the hydrolysis of protein. It is one of the most widely used enzyme preparations in the world. It can be used in detergent, leather, food, wine making industries, as well as in the production of textiles, pharmaceuticals, cosmetics, etc. Proteases are divided into Animal protease, plant protease and microbial protease. Proteases can also be divided into acid proteases, neutral proteases and alkaline proteases according to the optimum pH of the reaction. Alkaline protease is a serine proteolytic enzyme in endopeptidase, which is mainly used in the enzyme detergent industry and was first found in the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/54C11D3/386C12R1/07
CPCC12N1/20C12N9/54C11D3/38618C12R2001/07C12N1/205
Inventor 石亚伟文阳宣周桂旭
Owner SHANXI UNIV
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