Bacillus subtilis for highly producing D-ribose and co-producing acetoin
A technology of Bacillus subtilis and acetoin, which is applied to bacteria, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of high cost and low utilization rate of raw materials, and achieves high mutation rate, extensive culture conditions, Light damage effect
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Embodiment 1
[0042] Incline medium: 5g / L D-sorbitol, 10g / L peptone, 2g / L yeast extract, 2g / L NaCl, 20g / L agar, pH 6.9.
[0043] Seed medium: 20g / L glucose, 20g / L corn steep liquor, 3g / LK 2 HPO 4 , 1g / LKH 2 PO 4 , 1g / L yeast extract, pH 6.9.
[0044] Fermentation medium: 200g / L glucose, 8g / L corn steep liquor, 4g / L yeast powder, 7g / L (NH 4 ) 2 SO 4 , 0.05g / L MnSO 4 , 3g / LCaCO 3 , 1.5g / L sorbic acid, pH 6.9; 3L of fermentation broth in a 5L fermenter, sterilized at 121°C for 15 minutes.
[0045] Put two loops of Bacillus subtilis CGMCC No.3720 in the seed medium, cultivate at 36°C for 18 hours to obtain a mature seed liquid, and insert the seed liquid into the cooling medium according to the inoculation amount of 10% (v / v) of the fermentation liquid volume In the final fermentation broth, dissolved oxygen was controlled at 1.1vvm, 600rpm, and fermented at 37°C for 72 hours. The content of D-ribose and acetoin in the finally obtained fermentation broth were 113g / L and 30.5g / L respect...
Embodiment 2
[0051] Fermentation medium: 150g / L glucose, 50g / L xylose, 5g / L corn steep liquor, 6g / L yeast extract, 5g / L (NH 4 ) 2 SO 4 , 0.05g / L MnSO 4 , 2g / LCaCO 3 , 0.4g / L sorbic acid, pH 6.9; 3L of fermentation broth in a 5L fermenter, sterilized at 121°C for 15 minutes.
[0052] Prepare seed culture medium, formula and culture method are with embodiment 1, seed liquid is inserted in the fermentation culture liquid after cooling by the inoculum size of 10% (v / v) of fermented liquid volume, dissolved oxygen control is 1vvm, 500rpm, Under 37 ℃ of fermentation 72h, the D-ribose content in the fermented liquid obtained at last is 98.51g / L, and the acetoin content is 29.1g / L, respectively compared with only using common culture medium (that is, does not add sorbic acid, but other components The same) yield increased by 12.95% and 29.33%.
Embodiment 3
[0054] Fermentation medium: 20g / L glucose, 20g / L molasses, 1g / L corn steep liquor, 0.5g / L peptone, 0.5g / L yeast powder, 0.5g / L (NH 4 ) 2 SO 4 , 0.02g / L MnSO 4 , 0.01g / LMgSO 4 , 0.01g / LCaCO 3 , 3.5g / L sorbic acid, pH 6.9; 3L of fermentation broth in a 5L fermenter, sterilized at 121°C for 15 minutes.
[0055] Prepare seed culture medium, formula and culture method are with embodiment 1, seed liquid is inserted in the fermentation culture liquid after cooling by the inoculum size of 10% (v / v) of fermented liquid volume, dissolved oxygen control is 1vvm, 550rpm, Under 37 ℃ of fermentation 70h, the D-ribose content in the fermented liquid obtained at last is 91.4g / L, and the acetoin content is 25.5g / L, which are respectively compared with only using common culture medium (that is, without adding sorbic acid, but other components The same) yield increased by 4.80% and 13.33%.
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