Method for producing 2-phenethyl alcohol by utilizing mixed bacteria fermentation
A technology of mixed bacterial fermentation and phenylethanol, applied in the field of microbial fermentation, can solve the problems of heavy load of single bacteria, long metabolic path, low yield, etc., and achieve important application value and cost reduction effect.
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Embodiment 1
[0035] The impact of different sugar supplementation rates on the final 2-phenylethanol output of embodiment 1
[0036] (1) Escherichia coli was inoculated into the activation medium with an inoculation amount of 5 % v / v, and activated at 37°C and 300 rpm for 24 h;
[0037] The activation medium formula is: 10 g / L NaCl, 5 g / L yeast powder, 10 g / L peptone, the solvent is water, and the pH is adjusted to 6.8;
[0038] (2) Inoculate the activated Escherichia coli YLC20 into the fermentation medium with an inoculation amount of 5 % v / v, and ferment for 24 h at 37 °C;
[0039] Fermentation medium formula: 3g / L MgSO 4 , 3 g / L KH 2 PO 3 , 1 g / L NaCl, 5 g / L (NH 4 ) 2 SO 4 , 0.015 g / L CaCl 2 2H 2 O, 0.1125 g / L FeSO 4 ·7H 2 O, 1 g / L trisodium citrate, 10 g / L yeast powder, 0.3 g / L tyrosine, 0.15 g / L VB 1 , 0.5 g / L YNB, 20 g / L glucose, the solvent is water, adjust the pH to 6.0-7.0, and sterilize at 115°C for 20 min; set up three groups of fermentation experiments, in which the...
Embodiment 2
[0044] The impact of the different inoculation times of Pichia pastoris on the final 2-phenylethanol output in embodiment 2
[0045] The method is the same as in Example 1, wherein the rate of sugar supplementation is 4 g / L / h; the difference is that in step (2), 4 groups of experiments are set up, and the fermentation time is 12 h, 24 h, 36 h, and 48 h, respectively.
[0046] During the cultivation process, samples were taken every 12 h and the production of 2-phenylethanol was determined. When Escherichia coli YLC20 was cultured for 24 hours and Pichia mongolia was added, the production of 2-phenylethanol was the highest, reaching 1.75 g / L. In this system, the strain YLC20 can effectively use glucose to grow and obtain about 9 g / L of L-phenylalanine. After that, in the mixed bacteria system, Escherichia coli can still use glucose to grow continuously, and can continuously convert glucose into It is L-phenylalanine, and Pichia pastoris can use the transformed L-phenylalanine ...
Embodiment 3
[0047] Example 3: Pichia pastoris Meyerozyma guilliermondii Effects of different inoculum amounts of YLG18 on the yield of 2-phenylethanol
[0048] The method is the same as in Example 1, wherein the rate of sugar supplementation is 4 g / L / h, and the inoculation time is 24 h. The difference is that 4 groups of experiments are set up in step (3), and the inoculum amount of Pichia monsoonii in each group is 5 % v / v, 10% v / v, 15% v / v, 20% v / v.
[0049] During the cultivation process, samples were taken every 12 h and the production of 2-phenylethanol was determined. When the inoculation amount was 10% v / v, the 2-phenylethanol production was the highest, with 2.0 g / L; when the inoculum amount was 5% v / v, the strain grew slowly, which affected the production of 2-phenylethanol; while the inoculum amount When it is 15%-20% v / v, because too much inoculum will affect the synthesis of L-phenylalanine by Escherichia coli, 10% v / v inoculum is the optimal inoculum amount.
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