Saccharomyces cerevisiae and application thereof in preparing (R)-(+)-beta-hydroxy-benzenepropanoic acid ethylester by microbial transformation
A technology of ethyl carbonyl phenylpropionate and Saccharomyces cerevisiae, which is applied to Saccharomyces cerevisiae and the application field in the preparation of (R)-(+)-β-hydroxyphenylpropionate ethyl ester by microbial transformation, and can solve the problem of product yield and Problems such as unsatisfactory enantioselectivity, to achieve the effects of easy large-scale industrial production, high biotransformation rate, and simple production and operation
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Embodiment 1
[0027] Slant culture: Inoculate CGMCC No.3361 bacteria into the slant medium and culture at 30°C for 4-6 days. The slant medium is prepared as follows: wort juice 10g / L, yeast powder 3g / L, peptone 5g / L, glucose 10g / L, agar 20g / L, natural pH, solvent is water; sterilized at 121°C 20min, after sterilizing, cool to make a slope.
[0028] Seed culture and fermentation: Both the seed and the fermentation medium use liquid medium, which is prepared according to the following composition: glucose 30g / L, yeast powder 3g / L, ammonium sulfate 5g / L, anhydrous MgSO 4 0.25g / L, K 2 HPO 4 ·3H 2 O 1g / L, KH 2 PO 4 1g / L, sterilized at 121°C for 20min. The pH value of the liquid medium was adjusted to 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0 with 1mol / L NaOH or 1mol / L HCl respectively.
[0029] Use an inoculation needle to take a ring of bacteria from the slant culture medium and inoculate it into a 250ml Erlenmeyer flask containing 100ml of liquid medium, and culture it at 30°C and 1...
Embodiment 2
[0035] Slant culture: Inoculate CGMCC No.3361 bacteria into the slant medium and culture at 30°C for 4-6 days. The slant medium is prepared as follows: wort juice 10g / L, yeast powder 3g / L, peptone 5g / L, glucose 10g / L, agar 20g / L, natural pH, solvent is water; sterilized at 121°C 20min, after sterilizing, cool to make a slope.
[0036] Seed culture and fermentation: Both the seed and the fermentation medium use liquid medium, which is prepared according to the following composition: glucose 30g / L, yeast powder 3g / L, ammonium sulfate 5g / L, anhydrous MgSO 4 0.25g / L, K 2 HPO 4 ·3H 2 O 1g / L, KH 2 PO 4 1g / L, sterilized at 121°C for 20min. Adjust the pH of the liquid medium to 7.0.
[0037] Use an inoculation needle to take a ring of bacteria from the slant culture medium and inoculate it into a 250ml Erlenmeyer flask containing 100ml of liquid medium, and culture it at 30°C and 180r / min for 24h to obtain seeds. The seed liquid was inoculated into a 250 ml Erlenmeyer flask ...
Embodiment 3
[0043] Slant culture: Inoculate CGMCC No.3361 bacteria into the slant medium and culture at 30°C for 4-6 days. The slant medium is prepared as follows: wort juice 10g / L, yeast powder 3g / L, peptone 5g / L, glucose 10g / L, agar 20g / L, natural pH, solvent is water; sterilized at 121°C 20min, after sterilizing, cool to make a slope.
[0044] Seed culture and fermentation: Both the seed and the fermentation medium use liquid medium, which is prepared according to the following composition: glucose 30g / L, yeast powder 3g / L, ammonium sulfate 5g / L, anhydrous MgSO 4 0.25g / L, K 2 HPO 4 ·3H 2 O 1g / L, KH 2 PO 4 1g / L, sterilized at 121°C for 20min. Adjust the pH of the liquid medium to 7.0.
[0045] Use an inoculation needle to take a ring of bacteria from the slant culture medium and inoculate it into a 250ml Erlenmeyer flask containing 100ml of liquid medium, and culture it at 30°C and 180r / min for 24h to obtain seeds. The seed liquid was inoculated into a 250 ml Erlenmeyer flask ...
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