Method for producing sophorolipid through fermentation by taking cottonseed molasses and cottonseed oil as substrates
A technology of cottonseed molasses and microbial fermentation biosurfactant is applied in the field of microbial fermentation biosurfactant production to achieve the effects of improving utilization rate, simple process and reducing energy consumption
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Embodiment 1
[0032] (1) Replacement of the first carbon source and the second carbon source: prepare four sets of fermentation media (g / l): a glucose 80.0, oleic acid 6% (v / v); b glucose 80.0, cottonseed oil 6% (v / v); c cottonseed molasses 133.33 (sugar 80.0), oleic acid 6% (v / v), d cottonseed molasses 133.33 (sugar 80.0) cottonseed oil 6% (v / v), KH is added to each group 2 PO 4 1.0, Na 2 HPO 4 1.0, MgSO 4 ·7H 2 O0.50, yeast powder 3.0, natural pH.
[0033] (2) Fermentation to produce sophorolipids: Sterilize the fermentation medium of the fourth group in (1) at 115°C for 30 minutes and then cool. The activated Wickerhamierlla domercqiae var. sophorolipid CGMCC No. 1576 was inoculated into the above four fermentation media at an inoculum of 2% by volume at 25°C, 200 rpm, and shaker fermentation 7 Day, the fermentation broth of sophorolipid was obtained.
[0034] (3) Crude extraction of sophorolipid: take the fermentation broth containing sophorolipid and add 2 times its volume of ethyl acetat...
Embodiment 2
[0038] (1) Nitrogen source gradient design: prepare five groups of fermentation media (g / l): the addition of yeast powder is 0, 1.0g / l, 2.0g / l, 3.0g / l, 4.0g / l. Gradient, and add cottonseed molasses 133.33 (sugar 80.0), KH 2 PO 4 1.0, Na 2 HPO 4 1.0, MgSO 4 ·7H 2 O0.50, cottonseed oil 6% (v / v), natural pH.
[0039] (2) Fermentation to produce sophorolipids: Sterilize the five groups of fermentation medium (1) at 115°C for 30 minutes, and then cool. The activated Wickerhamierlla domercqiae var. sophorolipid CGMCC No. 1576 was inoculated into the above fermentation medium at an inoculum of 2% by volume at 25°C, 200 rpm, and shaker for 7 days. The fermentation broth of sophorolipid was obtained.
[0040] (3) Crude extraction of sophorolipid: take the fermentation broth containing sophorolipid and add 2 times its volume of ethyl acetate for extraction, centrifuge at 6000 rpm for 15 minutes, collect the ethyl acetate layer, concentrate under reduced pressure and dry to obtain the lacton...
Embodiment 3
[0044] (1) Influence of inorganic salt: prepare two sets of fermentation medium (g / l): one set is added with KH 2 PO 4 1.0, Na 2 HPO 4 1.0, MgSO 4 ·7H 2 O0.50, the other group does not add inorganic salt ions, both groups add cottonseed molasses 133.33 (sugar 80.0), cottonseed oil 6% (v / v), natural pH.
[0045] (2) Sophorolipid production by fermentation: The two sets of fermentation medium in (1) were sterilized at 115°C for 30 minutes and then cooled. The activated Wickerhamierlla domercqiae var. sophorolipid CGMCC No. 1576 was inoculated into the above fermentation medium at an inoculum of 2% by volume at 25°C, 200 rpm, and shaker for 7 days. The fermentation broth of sophorolipid was obtained.
[0046] (3) Crude extraction of sophorolipid: take the fermentation broth containing sophorolipid and add 2 times its volume of ethyl acetate for extraction, centrifuge at 6000 rpm for 15 minutes, collect the ethyl acetate layer, concentrate under reduced pressure and dry to obtain the l...
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