Synthesis of (R)-styrene glycol by coupling acceleration of (R)-carbonyl reduction enzyme and formic dehydrogenase
A technology of phenylethylene glycol and hydroxyacetophenone, which is applied in the field of biocatalytic asymmetric transformation, and can solve the problems of unstable coenzyme and high price
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Embodiment 1
[0068] For the cell culture of Candida parapsilosis (C.parapsilosis) CCTCC M203011 and Candida boidinii (C.boidinii), the culture medium consists of: glucose 4%, yeast extract 0.5%, (NH4) 2 HPO 4 1.3%, KH 2 PO 4 0.7%, ZnSO 4· 7H 2 O0.03%, NaCl 0.01%, pH7.0.
[0069] The two kinds of yeast were inoculated into 250 mL shake flasks with 20% medium content and cultured at 30° C. and 150 rpm for 48 h with shaking.
Embodiment 2
[0071] Extraction of Candida parapsilosis (C.parapsilosis) and Candida boidinii (C.boidinii) genome: the thalline cultured in Example 1 was centrifuged at 6,000 rpm for 20 min, washed twice with physiological saline, and collected Genome was extracted from cells using Genomic DNA Extraction Miniprep System (VIOGENE Company), a genomic DNA extraction kit.
Embodiment 3
[0073] Codon-optimized primer design for the rcr gene:
[0074] Rcr-F1: 5'-ATCG CATATG TCAATTCCATCAAGCCAGTACGGATTCGTATTCAATAAGCAATCAGGACTTAAGTTGCGT-3' (Nde I),
[0075] Rcr-F2: 5'-GCATTGCGTGAAATTCGTATCTTG-3',
[0076] Rcr-R1: 5'-CAAGATACGAATTTCACGCAATGC-3',
[0077] Rcr-R2: 5'-TGACT CTCGAG CTATGGATTAAAAACAACACGACCTTCATAAGCATTGTTACGCAA-3' (Xho I). When designing mutation points, the CGA or AGA at the 61st-63rd, 829th-831st, 838th-840th, 970th-972th, 991st-993rd were all mutated into CGT.
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