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A kind of separation method of bacillus amyloliquefaciens q-426 and lipopeptide thereof

A technology for the separation of amyloliquefaciens spores and methods, which is applied in the field of separation of Bacillus amyloliquefaciens Q-426 and its lipopeptides, which can solve problems such as inability to separate various lipopeptide components, and achieve broad-spectrum antifungal activity and nutritional requirements Not high, improve the effect of solvent utilization

Active Publication Date: 2020-04-24
DALIAN NATIONALITIES UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Generally speaking, for the separation and purification of lipopeptides, most of them only stay at the crude extraction at the laboratory level. The purification methods and means are generally a simple combination of acid precipitation and macroporous adsorption resin chromatography, and cannot simultaneously treat multiple lipids. Peptide fractions are separated

Method used

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  • A kind of separation method of bacillus amyloliquefaciens q-426 and lipopeptide thereof
  • A kind of separation method of bacillus amyloliquefaciens q-426 and lipopeptide thereof
  • A kind of separation method of bacillus amyloliquefaciens q-426 and lipopeptide thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] (1) Bacillus amyloliquefaciens Q-426 from China Type Culture Collection Center (CCTCC NO.M 2010237) was sequentially activated, first-level seed fermentation, and fermentation culture to obtain a fermentation liquid;

[0057] (2) Acid precipitation: Centrifuge 100 mL of the fermentation broth to remove bacteria, take the supernatant, adjust the pH of the supernatant to 2.5 for acid precipitation, and centrifuge (6000 rpm, 15 min) to remove the supernatant to obtain a precipitate;

[0058] (3) Cleaning: Use 20 mL of ultrapure water with a pH of 1.8 to wash the precipitate obtained in step (1), and wash twice;

[0059] (4) Extraction: use 40mL 100wt% methanol to resuspend the precipitate washed in step (2), shake it for 3h, centrifuge (12000rpm, 5min) to leave the supernatant, repeat the vibration of the precipitate 3 times with 100wt% methanol, and combine centrifuged supernatant after 3 extractions;

[0060] (5) Concentration: Concentrate the supernatant obtained in st...

Embodiment 2

[0065] (1) Bacillus amyloliquefaciens Q-426 from China Type Culture Collection Center (CCTCC NO.M 2010237) was sequentially activated, first-level seed fermentation, and fermentation culture to obtain a fermentation liquid;

[0066] (2) Acid precipitation: Centrifuge 100 mL of the fermentation broth to remove bacteria, take the supernatant, adjust the pH of the supernatant to 3.0 for acid precipitation, and centrifuge (6000 rpm, 15 min) to remove the supernatant to obtain a precipitate;

[0067] (3) Cleaning: Use 20 mL of ultrapure water with a pH of 1.8 to wash the precipitate obtained in step (1), and wash twice;

[0068] (4) Extraction: use 40mL 100wt% methanol to resuspend the precipitate washed in step (2), shake it for 3h, centrifuge (12000rpm, 5min) to leave the supernatant, repeat the vibration of the precipitate 3 times with 100wt% methanol, and combine centrifuged supernatant after 3 extractions;

[0069] (5) Concentration: Concentrate the supernatant obtained in st...

Embodiment 3

[0074] (1) Bacillus amyloliquefaciens Q-426 from China Type Culture Collection Center (CCTCC NO.M 2010237) was sequentially activated, first-level seed fermentation, and fermentation culture to obtain a fermentation liquid;

[0075] (2) Acid precipitation: Centrifuge 100 mL of the fermentation broth to remove bacteria, take the supernatant, adjust the pH of the supernatant to 2.0 for acid precipitation, and centrifuge (6000 rpm, 15 min) to remove the supernatant to obtain a precipitate;

[0076] (3) Cleaning: Use 20 mL of ultrapure water with a pH of 1.8 to wash the precipitate obtained in step (1), and wash twice;

[0077] (4) Extraction: use 40mL 100wt% methanol to resuspend the precipitate washed in step (2), shake it for 3h, centrifuge (12000rpm, 5min) to leave the supernatant, repeat the vibration of the precipitate 3 times with 100wt% methanol, and combine centrifuged supernatant after 3 extractions;

[0078] (5) Concentration: Concentrate the supernatant obtained in st...

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Abstract

The invention relates to the technical field of microbial metabolism products, in particular to bacillus amyloliquefaciens Q-426 and a lipopeptid separation method thereof. The bacillus amyloliquefaciens Q-426 can be cultivated easily and is low in nutritional requirement, capable of well growing within a wide temperature and pH value range, capable of achieving broad-spectrum antifungal activity and high in inhibiting effect on various tumor cells. The lipopeptid separation method mainly comprises the eight steps of fermental cultivation, acidic precipitation, cleaning, extraction, concentration, cation exchange resin adsorption, chromatographic analysis resin adsorption and desorption and semi-preparation type high performance liquid chromatograph (HPLC) collection. A set of method capable of separating various lipopeptid components synchronously is established, and the method has remarkable advantages compared with a previous active carbon adsorption method, a macroporous resin adsorption method and the column chromatography; the single components can be obtained, both the purity and the yield are high, and time needed by purification is shortened obviously.

Description

technical field [0001] The invention relates to a method for separating bacillus amyloliquefaciens Q-426 and lipopeptide, belonging to the technical field of microbial metabolites. Background technique [0002] Bacillus amyloliquefaciens is a species of Bacillus, which has a strong ability to produce secondary metabolites and can produce a variety of metabolites with biological activity. Lipopeptide is one of the important metabolites, which is composed of short peptides and Amphiphilic molecules composed of fatty acid chains: peptide chains composed of multiple amino acids form a hydrophilic group, and aliphatic hydrocarbon chains form a lipophilic group. Due to its special chemical structure, lipopeptide substances have various biological activities such as antibacterial, anti-inflammatory, anti-tumor, and anti-viral, and have the advantages of heat resistance, acid resistance, protease stability, low toxicity, and low drug resistance. [0003] In addition to its antibact...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C07K1/30C07K1/18C07K1/16C12R1/07
Inventor 权春善金黎明郑维周伟刘静范圣第
Owner DALIAN NATIONALITIES UNIVERSITY
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