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
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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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