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Production method of bio-surfactin

A biological surface and production method technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., to achieve the effects of avoiding the reduction of biosurfactant content, high product purity, and good extraction effect

Active Publication Date: 2016-06-22
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The object of the present invention is to provide a kind of production method of biosurfactant in order to solve above-mentioned deficiencies in the prior art, can realize industrialized large-scale production of high-purity biosurfactin, make up the deficiency that current biosurfactant cannot be produced on a large scale

Method used

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  • Production method of bio-surfactin
  • Production method of bio-surfactin
  • Production method of bio-surfactin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1: the influence of soya bean flour concentration on surfactin output

[0028] Cultivate Bacillus amyloliquefaciens fmb50 (preservation number CGMCCNo.6249) on PDA slant medium at 37°C for 24h, then inoculate a loop on BPY seed medium, culture at 37°C, 180rpm for 18h, inoculate at 3v / v% inoculum In the following medium with different carbon sources, culture at 33° C., 180 rpm for 36 hours.

[0029] Basic components: glucose 20g / L, K 2 HPO 4 ·3H 2 O9g / L, MgSO 4 ·7H 2 O0.6g / L, CaCl 2 0.1g / L, FeSO 4 ·7H 2 O0.01g / L, MnSO 4 ·H 2 O0.05g / L, urea 1g / L, pH7.0, where glucose is sterilized alone. Add one of 40g / L, 60g / L, 80g / L, 100g / L soy flour which is sterilized separately to the above composition. Measure the content of surfactin by HPLC (its standard liquid chromatogram sees figure 1 ), and use the external standard method to calculate the amount of surfactin produced by fermentation of soybean flour with different contents (surfactin standard curve see ...

Embodiment 2

[0032] Embodiment 2: the impact of carbon source on surfactin output

[0033] Cultivate Bacillus amyloliquefaciens fmb50 (preservation number CGMCCNo.6249) on PDA slant medium at 37°C for 24h, then inoculate a loop on BPY seed medium, culture at 37°C, 180rpm for 18h, inoculate at 3v / v% inoculum In the following medium with different carbon sources, culture at 33° C., 180 rpm for 36 hours.

[0034] Basic components: soybean powder 80g / L, K 2 HPO 4 ·3H 2 O9g / L, MgSO 4 ·7H 2 O0.6g / L, CaCl 2 0.1g / L, FeSO 4 ·7H 2O0.01g / L, MnSO 4 ·H 2 O0.05g / L, urea 1g / L, pH7.0.

[0035] To the above composition was added one of the following carbon sources which were sterilized separately.

[0036] 10g / L, 20g / L, 30g / L of molasses, glucose, soluble starch, maltose syrup, maltodextrin and combinations thereof.

[0037] The culture solution was centrifuged, and surfactin in the supernatant was quantified by the following HPLC method.

[0038] The amount of surfactin produced by fermentati...

Embodiment 3

[0041] Embodiment 3: the production of surfactin in 10L fermenter

[0042] Cultivate Bacillus amyloliquefaciens fmb50 (preservation number CGMCCNo.6249) on PDA slant medium at 37°C for 24h, then inoculate a loop on BPY seed medium, culture at 37°C, 180rpm for 16h, inoculate at 1.5v / v% inoculum In the fermentation medium, cultured at 33°C for 40h, the formulation of the fermentation medium is: soybean powder 80g / L, maltodextrin 20g / L, K 2 HPO 4 ·3H 2 O9g / L, MgSO 4 ·7H 2 O0.6g / L, CaCl 2 0.1g / L, FeSO 4 ·7H 2 O0.01g / L, MnSO 4 ·H 2 O0.05g / L, urea 1g / L, emulsified silicone oil 0.5mL / L, pH7.0, among which maltodextrin was added after sterilization alone. After inoculation, adjust the air flux to 1VVM, adjust the dissolved oxygen to 100% when the speed is 180rpm, maintain the air flux to 1VVM, maintain the speed between 180 and 600rpm and keep DO>20%, during which the reduction in the fermentation broth is measured by DNS method Sugar content, when the fermentation culture r...

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Abstract

The invention discloses a production method of bio-surfactin. The method comprises the following steps of: sequentially inoculating bacillus amyloliquefaciens fmb50 to a PDA agar slant culture medium for culture, then inoculating the bacillus amyloliquefaciens fmb50 to a BPY seed culture medium for further culture, and finally inoculating the bacillus amyloliquefaciens fmb50 to a fermentation culture medium, thereby obtaining a culture solution containing bio-surfactin; then diluting the culture solution with water, then regulating the pH, further adding chitosan, sodium alga acid and kieselguhr, performing flocculation, and then filtering; finally dissolving and extracting a filter cake with low alcohols, then filtering, and performing reduced-pressure distillation on the filter liquor, thereby obtaining a bio-surfactin crude product. The production method of bio-surfactin, provided by the invention, is capable of realizing industrial preparation of bio-surfactin; the prepared bio-surfactin is high in purity, so that the method is capable of overcoming the defect that large-scale production of bio-surfactin can not be realized in the prior art.

Description

technical field [0001] The invention relates to a method for fermenting and isolating surfactin with microorganisms belonging to the genus Bacillus, in particular to a production method for biological surfactin. Background technique [0002] Surfactin is a cyclic peptide synthesized by Bacillus amyloliquefaciens. Due to the presence of hydrophilic peptide bonds and hydrophobic fatty acid chains in its molecular structure, it has extremely strong surface activity. Surfactin can interact with the cell membrane, causing the perforation of the cell membrane and the leakage of cytoplasm, thereby interfering with the normal cell structure. In addition to its high surface activity, Surfactin has also been proven to have anti-tumor, anti-mycoplasma and anti-virus effects, so it is widely used in the fields of medicine, food, agricultural biocontrol and breeding. [0003] During the fermentation process of Surfactin, due to its strong surface activity, the culture solution will foam...

Claims

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Application Information

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IPC IPC(8): C12P21/04C12R1/07
CPCC07K7/50
Inventor 陆兆新唐伟吕凤霞别小妹赵海珍张充
Owner NANJING AGRICULTURAL UNIVERSITY
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