Aeromonas and application of aeromonas in biotransformation of (R)-alpha-hydroxyphenylacetic acid
A hydroxyphenylacetic acid, Aeromonas guinea pig technology, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of unavailability of S enantiomer, low yield, environmental pollution, etc. Low production cost, high efficiency and less pollution
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[0039]The application of Aeromonas caviae B18 in the asymmetric conversion of (R)-alpha-hydroxyphenylacetic acid is to prepare (R)-alpha-hydroxyphenylacetic acid, and the preparation method step is to transform Aeromonas caviae Caviae B18 is transferred to the transformation medium with an inoculum of 4% by volume, and the substrate acetophenone is added to the transformation medium, and the transformation culture is based on the conditions of 30-36°C and the shaker speed is 200r / min. For 19 hours, the pH of the transformation medium was 7.2, and the bacterial cells were removed by centrifugation to obtain (R)-α-hydroxyphenylacetic acid fermentation broth. Under the conditions of microwave vacuum drying for 1 h, the purity of (R)-α-hydroxyphenylacetic acid was 99.5%.
[0040] The preparation method of (R)-α-hydroxyphenylacetic acid transformation medium is as follows: starch 12.81g, casein peptone 12.74g, KH 2 PO 4 2.97g, K 2 HPO 4 1.2g, MgSO 4 0.15g, CaCl 2 0.125g, MnSO...
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[0044] Screening of strains
[0045] Suspend 1g of pond bottom mud in 10mL of sterile water, carry out gradient dilution, and take 0.2mL of the diluted solution and spread it on a selective plate (including medium: KH 2 PO 4 1g, NaCl1g, K 2 HPO 4 1g, MgSO 4 0.15g, CaCl 2 0.125g, MnSO 4 0.015g, ZnSO 4 0.01g, substrate acetophenone acid 5mM, dilute to 1L with tap water, pH7.2). Add 50U / mL nystatin to the selective plate, incubate at 32°C for 2-3 days, observe the growth of the bacteria, and separate the bacteria group by streaking. Use a sterile toothpick to pick colonies of different shapes, and insert them into 2 mL of fermentation medium (starch 12.81 g, casein peptone 12.74 g, KH 2 PO 4 2.97g, K 2 HPO 4 1.2g, MgSO 4 0.15g, CaCl 2 0.125g, MnSO 4 0.015g, ZnSO 4 0.01g, pH7.2) in a test tube, cultured at 32°C, 200r / min shaker for 19h, added substrate acetophenone 5mM, transformed for 20h, and centrifuged the transformed fermentation broth at 10,000rpm for 10min, c...
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