Magnetic immobilization of yeast cells and its application in the preparation of ethyl (r)-2-hydroxy-4-phenylbutyrate
A technology of ethyl phenylbutyrate and immobilized yeast, which is applied in the direction of immobilized on or in the inorganic carrier, fermentation, oxides of ferrous iron, etc., can solve the problem of low resolution yield of chloramphenicol intermediates, The sample has to be diluted, the loss of carrier recovery is large, etc., to achieve the effect of improving biotransformation efficiency, enhancing tolerance, and good catalytic activity
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Embodiment 1
[0027] Embodiment 1: magnetic nanometer iron tetroxide (Fe 3 o 4 ) preparation
[0028] Weigh 12.675g FeCl 2 Dissolved in 1000mL deionized water, configured as 0.1M FeCl 2 solution. Weigh 16.220g FeCl 3 Dissolved in 1000mL deionized water, configured as 0.1M FeCl 3 solution. FeCl 2 Solution 30mL, FeCl 3 Mix 60mL of the solution into a 250mL three-neck flask, add 20mL of deionized water, and stir at a high speed of 1000 rpm, keep a water bath at 30-40°C, slowly add ammonia water dropwise to adjust the pH to 10, and the solution turns black and shiny. Raise the temperature to 60°C and react for 1h. Then raise the temperature to 80°C and ripen for 1h. After the reaction was finished, it was cooled to room temperature, and it was repeatedly washed with deionized water until it was colorless and transparent, and the pH of the supernatant was neutral, and collected by an external magnetic field to obtain a black magnetic particle precipitate (Fe 3 o 4 ), pre-frozen in -8...
Embodiment 2
[0029] Embodiment 2: the magnetic nano ferric oxide (Fe) of amino acid modification 3 o 4 - Preparation of AA)
[0030] Accurately weigh 0.2g each of the four amino acids (lysine, arginine, glutamic acid, and aspartic acid) and dissolve them in 200mL of deionized water. After shaking well, add 0.2g of magnetic Fe 3 o 4 After the nanoparticles were shaken properly, they reacted under the action of 40KHz ultrasonic wave for half an hour, took out the Erlenmeyer flask, collected the black precipitate with an external magnetic field, washed it with deionized water for 3 times, and then put it in a -80°C refrigerator for 8 hours. into a freeze dryer for 12 hours to obtain amino acid-modified Fe 3 o 4 Nanoparticles (denoted as Fe 3 o 4 -AA) 0.2-0.4 g. Among them, the lysine-modified magnetic nano-ferric iron tetroxide (Fe 3 o 4 -Lys) 0.35g, Fe 3 o 4 -Arg 0.27g, Fe 3 o 4 -Glu 0.23g, Fe 3 o 4 -Asp 0.21g. Identification of Fe by Fourier Transform Infrared Transform Spec...
Embodiment 3
[0031] Embodiment 3: the screening of active dry yeast
[0032] Weigh 3g of the purchased 11 kinds of active dry yeast and add them to 11 Erlenmeyer flasks filled with 20mL phosphate buffer (pH 7), shake well, and after fully dissolved, add them to the 11 kinds of 20mL yeast cell solutions respectively. 1mL of 0.2mmol / mL OPBE ethanol solution was placed in a shaker at 30°C and 180r / min for 24h. After the reaction was completed, the transformation solution was taken out and centrifuged at 8000rpm for 10min. The supernatant was collected and washed with an equal volume of Extracted with ethyl acetate, naturally volatilized and concentrated to a volume of 3 mL to make a sample to be tested by gas phase, the results are shown in Table 1, and No. 7 active yeast was selected for subsequent tests, that is, Baizuan high-activity dry yeast of Angel Yeast Co., Ltd. ( high sugar tolerance).
[0033] Gas chromatography (GC) detection conditions: chiral chromatographic column CP7502 (25m×...
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