Method for directional accumulation of specific homolog components of antimicrobial lipopeptides of bacilli

An antibacterial lipopeptide and bacillus technology, applied in the field of fermentation, can solve the problems of narrow antibacterial spectrum, single component, single function of lipopeptide products, etc., to achieve the effect of improving efficacy and expanding the scope of use

Inactive Publication Date: 2016-06-22
GUANGDONG OCEAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most studies on the production methods of antimicrobial lipopeptides are mainly carried out on the yield of lipopeptides, and the composition of homologues of lipopeptides is rarely considered, which makes the functions of the lipopeptide products produced relatively single
Under certain fermentation conditions, although the total yield of lipopeptides produced by the strain is high, the components of the lipopeptides produced are relatively single, which makes the antibacterial spectrum of the product relatively narrow, which affects the full play of the lipopeptides.

Method used

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  • Method for directional accumulation of specific homolog components of antimicrobial lipopeptides of bacilli
  • Method for directional accumulation of specific homolog components of antimicrobial lipopeptides of bacilli
  • Method for directional accumulation of specific homolog components of antimicrobial lipopeptides of bacilli

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Experimental program
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Embodiment 1

[0030] The BPY medium was used as the seed medium, and the seed solution was obtained by culturing for 18-24 hours. For liquid fermentation, the seed liquid was added to Landy medium at 5% (v / v), and the volume of the liquid was 80mL / 250mL in a Erlenmeyer flask. At 28°C, when the shaking table rotated at 160r / min, cultured for 48h, and the fermentation liquid was placed in Centrifuge at 10,000rpm for 15min, take the supernatant, adjust the pH to 2 with hydrochloric acid, let stand overnight, centrifuge at 10,000rpm for 20min, take the precipitate, dissolve it with methanol, adjust the pH to 7.0, centrifuge at 10,000rpm for 20min, take the supernatant and use rotary evaporation The instrument was concentrated at 60°C, and extracted with methanol three times. After concentration, the sample was placed in a refrigerator at 4°C for later use to obtain the fermentation extract.

[0031] The yield of lipopeptide obtained after detection was 723.82mg·L -1, the produced Iturin compon...

Embodiment 2

[0033] The inoculum size was the same as in Example 1, using Landy medium, at 30°C, at a shaker speed of 160r / min, cultured for 48h, and the output of lipopeptide was 763.20mg L -1 , the produced Iturin component accounts for 74.32% of the total mass, and Iturin is mainly composed of homologue components with mass-to-core ratios of 1044.30, 1057.20 and 1071.20, and the proportions of each homologue in the Iturin component are 2.40%, 3.67%, 93.93%. The content of Surfactin components accounted for 25.68%, mainly composed of homologues with mass-to-core ratios of 995.20, 1008.20, 1022.30 and 1058.20, and the proportions of each homologue in Surfactin components were 28.57%, 23.53%, 32.05% and 15.85% respectively .

Embodiment 3

[0035] The amount of inoculum was the same as in Example 1, using Landy medium, at 32°C, at a shaker speed of 160r / min, cultured for 48h, and the output of lipopeptide was 872.01mg L -1 , the produced Iturin component accounted for 71.46% of the total mass, and Iturin was mainly composed of homologue components with mass-to-core ratios of 1044.30, 1057.20 and 1071.20, and the proportions of each homologue in the Iturin component were 4.92%, 9.59%, 85.49%. The content of Surfactin component accounts for 28.54%, mainly composed of homologues with mass-to-core ratios of 995.20, 1008.20, 1022.30 and 1058.20, and the proportions of each homologue in Surfactin components are 31.28%, 22.05%, 30.54% and 16.13% respectively .

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Abstract

The invention belongs to the technical field of fermentation and discloses a method for directional accumulation of specific homolog components of antimicrobial lipopeptides of bacilli. Bacillus natto NT-6 strains serve as original strains, Landy serves as a culture medium, culture is performed at 28-32 DEG C for 36-48 h under the condition of 160 r/min, and the specific components comprise Iturins containing homologs with cytoplasmic-nuclear ratios being 1044.30, 1057.20 and 1071.20 and Surfactins containing homologs with cytoplasmic-nuclear ratios being 995.20, 1008.20, 1022.30 and 1058.20. With the adoption of the method, directional accumulation can be performed on the specific homolog components of the antimicrobial lipopeptides of the bacilli by controlling the fermentation culture medium and fermentation conditions, lipopeptide products comprising different specific homolog components and having specific ratios of the specific homolog components can be produced, so that effects of the lipopeptide products are improved, and the use range of the lipopeptide products is expanded.

Description

technical field [0001] The invention relates to the technical field of fermentation, and more specifically relates to a method for directional accumulation of specific homologue components of bacillus antibacterial lipopeptides. Background technique [0002] Antimicrobial lipopeptides are generally metabolites produced by Gram-positive bacillus. The antimicrobial lipopeptides discovered and studied so far mainly focus on surfactin, fengycin, and iturin. The molecules of these lipopeptides are composed of two parts: hydrophilic peptide bonds and lipophilic aliphatic hydrocarbon chains. Due to their special chemical composition and amphiphilic molecular structure, lipopeptides have surface activity, foaming properties, anti-bacteria, Anti-fungal, anti-viral, anti-protozoal, anti-tumor and other important biological activities, it shows broad market development value, and has important applications in the fields of medicine, agriculture, food, cosmetics, biological control, oil...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/00C12R1/07
CPCC12P21/00
Inventor 孙力军孙东方王雅玲邓旗徐德峰刘颖
Owner GUANGDONG OCEAN UNIVERSITY
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