Phosphate → ethyl N-Boc-2-amino-3-cyclohexyl-3-hydroxypropionate

関与物質

反応手順

The reducing activity against ethyl N-Boc-2-amino-3-cyclohexyl-3-oxopropionate was calculated in the following manner. First, an appropriate amount of a crude enzyme solution and 100 mM phosphate buffer (pH 6.5) were added to a test tube to make a total volume of 0.5 ml. Further, 0.25 mg of ethyl N-Boc-2-amino-3-cyclohexyl-3-oxopropionate and 1 mg of NADH were added, and the reaction was allowed to proceed at 30° C. for 2 hours with shaking. After the reaction, 1 ml of ethyl acetate was added and extraction was carried out. The quantitation of the product ethyl N-Boc-2-amino-3-cyclohexyl-3-hydroxypropionate was carried by capillary gas chromatography (column: GL Sciences InertCAP 5 (ID 0.25 mm×30 m), column temperature: 200° C.; carrier gas: helium (70 kPa), detection: FID). The enzyme activity was calculated from the amount of the ethyl N-Boc-2-amino-3-cyclohexyl-3-hydroxypropionate formed. The activity such that 1 μmol of ethyl N-Boc-2-amino-3-cyclohexyl-3-hydroxypropionate is formed in 1 minute under the reaction conditions mentioned above is defined as 1 unit.

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