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Bacillus licheniformis CPL618 and screening and application thereof

A technology of Bacillus licheniformis and CPL618, applied in the field of microorganisms, can solve the problems of inability to produce high bacitracin and high cost, and achieve the effects of low cost and increased yield

Active Publication Date: 2021-12-21
HUAIYIN INSTITUTE OF TECHNOLOGY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The fermentation methods described in the current literature and patents are generally mixed fermentations of multiple carbon sources and nitrogen sources, such as the fermentation medium of CN109439581A, CN113234631A, etc., all of which are composed of two or more nitrogen sources, and the cost is relatively high, and Inability to produce high levels of bacitracin A

Method used

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  • Bacillus licheniformis CPL618 and screening and application thereof
  • Bacillus licheniformis CPL618 and screening and application thereof
  • Bacillus licheniformis CPL618 and screening and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Isolation, screening and antibacterial activity determination of Bacillus licheniformis CPL618 in soil:

[0030] It is obtained by separating from wheat field soil in Huaian City, and the separation process is as follows: select about 10g of soil at a depth of 10-20cm, add 100ml of normal saline and mix well. Water bath at 80°C for 1h, then add 4g NaCl, and bathe at 37°C for 2h. Fully soak about 18g of the soil after the water bath in 50mL of sterile water, take 1mL of the soil suspension and dilute it 10 times with sterile water, take 100uL of the diluted test solution and add it dropwise on the LB solid medium, spread evenly, and invert at 37°C Incubate overnight. The morphology of individual colonies on the medium was observed through aseptic operation, including the color, size, edge of the colony and surface roughness of the colony, and 53 strains for primary screening were obtained. The cultured primary screening strain was used as the test bacteria, Staphylococ...

Embodiment 2

[0033] Identification and whole genome sequencing of strain CPL618:

[0034] The screened strain CPL618 was cultivated to the logarithmic growth phase, and its genomic DNA was extracted. The 16srDNA fragment of the gene was amplified by the universal primers of the bacteria, sequenced, and the 16srDNA fragment of the gene was compared by using BLAST analysis method. The full length of the sequence is 1555bp. The sequence of the bacterial strain CPL618 of the present invention is compared with the sequences such as Bacillus licheniformis A4-3 registered by NCBI, and the homology is 100%, and the gene construction phylogenetic tree is as follows: image 3 As shown, combined with the results of physiological and biochemical experiments on the strain CPL618, it was identified as Bacillus licheniformis (Bacillus licheniformis) and named as Bacillus licheniformis CPL618. The deposit number is CCTCC NO: M20211158.

[0035] The screened strain CPL618 was sent to the Beijing Genome I...

Embodiment 3

[0037] Antibacterial effect of Bacillus licheniformis CPL618 on Staphylococcus aureus, Escherichia coli, Penicillium brevidis and yeast:

[0038] Preparation of medium and activation of indicator bacteria: Staphylococcus aureus and Escherichia coli use LB medium as the growth medium; Penicillium brevidis uses PDA medium as the growth medium; yeast uses MRS medium as the growth medium. Single colonies of Staphylococcus aureus, Escherichia coli, Penicillium breviscellus and yeast were cultured to the logarithmic phase in the corresponding liquid medium.

[0039] The antibacterial activity was measured by the coating method, and the specific steps included: taking a single colony of CPL618 strain in LB liquid medium and cultivating it at 37°C until the OD value was 0.8-0.9, then using Staphylococcus aureus, Escherichia coli, Penicillium breviscellus and yeast are indicator bacteria, take 0.2ml indicator bacteria solution and spread it evenly on the corresponding growth plate, div...

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Abstract

The invention discloses a bacillus licheniformis CPL618 as well as screening and application thereof. The strain CPL618 is identified as bacillus licheniformis, the strain CPL618 is preserved in the China Center for Type Microbiological Culture Collection (CCTCC), the preservation time is September 13, 2021, and the preservation number is CCTCC NO: M20211158. The bacillus licheniformis CPL618 disclosed by the invention has broad-spectrum antibacterial activity, and can antagonize the growth of gram-positive bacteria, gram-negative cocci and fungi. The invention provides an optimized industrial culture medium which can improve the yield of bacitracin A at lower cost, and the maximum yield can reach 2.3 g / L. The bacillus licheniformis disclosed by the invention has a wide utilization prospect in the aspects of preparation of antibacterial agents and production of bacitracin A.

Description

technical field [0001] The invention belongs to microorganisms, in particular to a bacillus licheniformis CPL618 and its screening and application. Background technique [0002] Bacillus licheniformis is a Gram-positive thermophilic bacterium, a typical facultative anaerobic microorganism, which can ferment glucose and other available carbon sources to produce overflow metabolites such as acetic acid. The cells are rod-shaped and solitary. When cultured in gravy medium, the colonies appear flat, ragged, white, rough, and wrinkled. Bacillus licheniformis can produce a variety of antibiotics such as peptides and phospholipids, which can have a good inhibitory effect on human pathogenic bacteria. Because Bacillus licheniformis is not pathogenic and can secrete proteins and some enzymes outside the cells, it is widely used in industrial production. [0003] Bacillus licheniformis is a kind of microorganism with important use value and economic value. It is rich in species, wi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02C12P21/02C07K7/58A01N63/22A01P1/00A01P3/00C12R1/10
CPCC12N1/20C12N1/02C12P21/02C07K7/58A01N63/22Y02A50/30
Inventor 罗楚平陈梅琳罗科程李彬陈新尹秀莲王小花田宝霞张金峰
Owner HUAIYIN INSTITUTE OF TECHNOLOGY
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