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Novel antimicrobial lipopeptide [delta Cl] Plipastatin and producing strain and application thereof

A technology of antibacterial lipopeptide and antibacterial substances, which is applied in the field of microbial fermentation to produce new antibacterial lipopeptides, which can solve the problems of separation and purification difficulty

Inactive Publication Date: 2016-04-20
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the molecular weight of the complex enzyme is between 100,000 and 1 million Daltons, it is difficult to separate and purify

Method used

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  • Novel antimicrobial lipopeptide [delta Cl] Plipastatin and producing strain and application thereof
  • Novel antimicrobial lipopeptide [delta Cl] Plipastatin and producing strain and application thereof
  • Novel antimicrobial lipopeptide [delta Cl] Plipastatin and producing strain and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0041] Example 1: Construction of the novel antimicrobial lipopeptide [ΔC1] Plipastatin-producing strain B.subtilisPB2-LC1 using Bacillus subtilis B.subtilisPB2-L as the starting strain.

[0042] 1 Materials and methods

[0043] 1.1 Materials

[0044] 1.1.1 Strains, plasmids

[0045] Escherichiacoli DH5α was purchased from Invirtogen Company, B.subtilisPB2-L strain was preserved in China General Microorganism Culture Collection Center (CGMCC), preservation number: CGMCCNO.11723, the complete sequence of temperature-sensitive plasmid pKS2 is shown in SEQ ID NO.1, derived from the following References: Yang Huilin, Wang Kun, et al. Knockout of Bacillus amyloliquefaciens ptsGHI gene and growth characteristics of defective strains [J]. Journal of South China University of Technology, 2012, 40, 95-100.

[0046] 1.1.2 Main reagents and instruments

[0047] Restriction enzymes, T4 DNA ligase and PMD-19T vector were purchased from TaKaRa Company; antibiotics ampicillin (Amp), kanam...

Embodiment 2

[0063] 1 The fermentation and separation and identification methods of [△C1] Plipastatin produced by Bacillus subtilis PB2-LC1 of the present invention are as follows:

[0064] 1.1 Fermentation culture for lipopeptide production

[0065] Inoculate the strain B.subtilisPB2-LC1 on the slant of the test tube into a conical flask equipped with a seed medium (agar removed from the nutrient agar medium), and cultivate it at 37°C and 130-150r / min for 24h, until it reaches the logarithmic growth phase. Concentration is 10 8 -10 9 CFU / L of seed solution. The B. subtilis PB2-LC1 seed solution prepared above was inoculated in the Landy fermentation medium at a concentration of 3-5%, and cultured at 33° C. and 180 r / min for 72 hours to obtain an antibacterial substance fermentation broth.

[0066] Take the fermented liquid prepared above and centrifuge at 5000r / min for 25min to remove bacteria, adjust the pH of the supernatant to 2.0, keep overnight at low temperature, centrifuge at 5000...

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Abstract

The invention discloses novel antimicrobial lipopeptide [delta Cl] Plipastatin and a producing strain and application thereof. The antimicrobial substance [delta Cl] Plipastatin is produced by utilizing bacillus subtilis PB2-LCl, wherein the antimicrobial substance is from B.subtilis PB2-LCl and is a generic term of a series of linear antimicrobial lipopeptide compounds which comprise seven components, and it is identified through LC-FTICR-MS that the molecular weights of the seven components are 980 Da, 994 Da, 1,008 Da, 1,022 Da, 1,036 Da, 1,050 Da and 1,064 Da. The antimicrobial lipopeptide compounds have inhibiting effects on escherichia coli, micrococcus luteus, penicillium notatum and peach bacteria erwinia and can prevent and control various plant diseases.

Description

technical field [0001] The invention belongs to a novel antibacterial lipopeptide produced by microbial fermentation, and relates to a novel antibacterial lipopeptide [△C1]Plipastatin, its producing bacteria and its application. Background technique [0002] The food fermentation industry, agricultural production and pharmaceutical industry have been facing the following problems for a long time: microbial contamination of food, plant diseases and insect pests, and the increasing resistance of pathogenic bacteria. Although traditional solutions (chemical preservatives, chemical pesticides, antibiotics) can temporarily alleviate the contradiction, however, with the advancement of detection technology and the improvement of environmental awareness, people began to pay attention to the potential harm of the above-mentioned chemicals. The development of high-efficiency, safe, non-toxic, stable performance, and broad-spectrum natural preservatives has become a research and applic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/75C12P21/02C07K14/32C07K1/16A01P1/00A01P3/00C12R1/125
CPCC07K14/32C12N15/75C12N2800/101
Inventor 别小妹刘洪霞陆兆新吕凤霞张充赵海珍
Owner NANJING AGRICULTURAL UNIVERSITY
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