Bacillus velezensis LfF-1 strain and application thereof in protease production

A technology of Bacillus velesi and Bacillus, applied in the field of microorganisms, can solve the problems of low activity of a single protease, a single type of bacteria, and a low conversion rate of polypeptides, and achieves wide application prospects, the method is simple and feasible, and the production efficiency is improved. Effect

Active Publication Date: 2019-12-10
LINGNAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The technical problem to be solved by the present invention is to overcome the relatively single types of strains used for protease fermentation production, the low activity of a single protease, the low hydrolysis efficiency and recovery rate of substrate proteins, the low conversion rate of polypeptides, and the existing The peptide bond selectivity of protease preparation tends to be homogeneous and other defects and deficiencies, providing a strain of Bacillus Velez LfF-1 and its application in the production of protease

Method used

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  • Bacillus velezensis LfF-1 strain and application thereof in protease production
  • Bacillus velezensis LfF-1 strain and application thereof in protease production
  • Bacillus velezensis LfF-1 strain and application thereof in protease production

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] The isolation and identification of embodiment 1 Veles bacillus LfF-1 bacterial strain

[0060] The medium formula used in this embodiment is as follows:

[0061] Liquid enrichment medium: yeast powder 0.5%, peptone 1.0%, NaCl 0.5%, agar 1.5%-2.0%, pH 7.0-7.2, sterilized at 121°C for 20 minutes;

[0062] Primary screening medium: 4.0% skimmed milk powder, 2.0% sucrose, 1.5%-2.0% agar, natural pH, sterilized at 105°C for 20min.

[0063] Liquid seed medium: yeast powder 0.5%, peptone 1.0%, NaCl 0.5%, pH 7.0-7.2, sterilized at 121°C for 20 minutes;

[0064] Re-screening (fermentation) medium: 4.0% sucrose, 2.0% peptone, 0.3% dipotassium hydrogen phosphate, 0.3% calcium carbonate, 0.1% Tween-80, pH 7.5, sterilized at 121°C for 20 minutes;

[0065] Slant medium: 0.5% yeast powder, 1.0% peptone, 0.5% NaCl, 1.5%-2% agar, pH 7.0-7.2, sterilized at 121°C for 20 minutes.

[0066] 1. Strain isolation and screening

[0067] 1. Enrichment culture

[0068] Take 10 g (or 10 mL) o...

Embodiment 2

[0105] The optimization of embodiment 2 Veles bacillus LfF-1 bacterial strain fermentation production protease process conditions

[0106] The medium formula used in this embodiment is as follows:

[0107] Liquid seed medium: tryptone 1%, yeast extract 0.5%, sodium chloride 1%, pH 7.2, sterilized at 121°C for 20 minutes;

[0108] Basic fermentation medium: 4.0% sucrose, 2.0% peptone, 0.3% dipotassium hydrogen phosphate, 0.3% calcium carbonate, pH 7.5, sterilized at 121°C for 20 minutes.

[0109] 1. Seed cultivation

[0110] Pick 1 loop of LfF-1 strain from the slant of the test tube, transfer it to 50mL seed medium, and then place it on a shaking table at 35°C and 200r / min for 24h to obtain LfF-1 liquid seeds.

[0111] 2. Liquid fermentation

[0112] Prepare the fermentation medium according to the test design, draw 2mL of LfF-1 liquid seeds under sterile conditions and transfer it to a conical flask containing 50mL of fermentation medium, and then place the conical flask o...

Embodiment 3

[0191] The preparation of embodiment 3 Veles bacillus LfF-1 bacterial strain protease preparation

[0192] 1. Preparation of protease solution

[0193] Adopt fed-batch fermentation process, 5 liters of full-automatic fermenters to prepare protease liquid, concrete experimental method is as follows:

[0194] Fermentation medium: 4% lactose, 2% beef extract, 0.5% KH 2 PO 4 , 0.09% Tween-80, 3% bran, pH 7.5.

[0195] Fermentation condition setting: initial stage of fermentation (0~24h), temperature is 33°C, sterile air flow rate is 125L / h, stirring speed is 180r / min, the foam level in the fermenter is controlled by automatic dropwise addition of Great Defoamer;

[0196] In the middle and late stage of fermentation (24h~60h), the temperature is 27°C, the sterile air flow rate is 210L / h, and the stirring speed is 350r / min. The flow rate adds 2L of sterilized fermentation medium to the fermenter (filtering removes the bran residue of the original medium).

[0197] After the fer...

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Abstract

The invention discloses a Bacillus velezensis LfF-1 strain and application thereof in protease production. The Bacillus velezensis LfF-1 strain is separated for a first time, the strain is preserved in the Guangdong Microbiological Culture Collection Center in August 16, 2019, and the preservation number is GDMCC NO: 60741. The strain has a remarkable capability for producing protease through fermentation, and types of cultures produced through protease fermentation can be enriched; a protease preparation can be produced by using the strain through a fermentation tank replenishing on-batch fermentation process, the activity of the protease is as high as 17.84 ten thousand U/g, the enzyme activity recovery rate is as high as 96.8%, types of conventional commercial protease can be greatly enriched, and the production efficiency of the protease preparation can be improved; and in addition, the method for producing the protease from the strain through fermentation is simple and feasible, and has wide application prospects in producing protein foods, functional peptide products or feed additives.

Description

technical field [0001] The invention belongs to the technical field of microorganisms. More specifically, it relates to a strain of Bacillus Velez LfF-1 and its application in the production of protease. Background technique [0002] Protease is a general term for a class of hydrolytic enzymes that can hydrolyze proteins and convert them into peptides and amino acids. They are widely used in daily chemicals, food processing, feed production and breeding. Compared with the early protein acid hydrolysis process, the use of protease to realize proteolysis is a clean and green production process with more development potential. In recent years, as the physiological and health functions of peptides have been continuously recognized by people, peptides have been widely used as raw materials in industries such as health food, clinical food and feed additives. [0003] Protease is an important digestive enzyme in various organisms, and it participates in a series of physiological ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/54C12R1/07
CPCC12N9/54C12N1/205C12R2001/07
Inventor 潘进权周鲜娇
Owner LINGNAN NORMAL UNIV
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