21880-87-5

530-93-8

2217-42-7

2217-42-7

The generalized procedure for the synthesis of the target compounds from β-tetralone was as follows: the final concentration of the reaction system was 0.25 μL, which was adjusted using 100 mM potassium phosphate (KPi) buffer at pH 7.5, and an equimolar amount of 70 mM of the amino donor was reacted with 70 mM of the ketone substrate in 0.1 mL of the cell free extract (CFE) containing the transcripts for 24 hours at 28°C. Specific operations include: co-warming CFE containing transaminases XP-001209325, AAN21261, and ABN35871, respectively, with benzylacetone and α-methylbenzylamine to form 4-phenyl-2-butylamine and acetophenone; reaction of phenylacetone and α-methylbenzylamine to form α-ethylbenzylamine and acetophenone; reaction of 1-indanone and α-methylbenzylamine to form 1-aminoindan and acetophenone; the reaction of 1-tetralone and α-methylbenzylamine to form 1-aminotetralone and acetophenone; the reaction of 2-tetralone and α-methylbenzylamine to form 2-aminotetralone and acetophenone; the reaction of butanone and α-methylbenzylamine to form 2-amino-butane and acetophenone; and the reaction of 3,3-dimethyl-2-butanone and α-methylbenzylamine to form, respectively, 3,3-dimethyl-2-amino-butane and acetophenone. Reaction termination was achieved by adding 0.75 mL of termination reagent (50% (v/v) aqueous acetonitrile solution containing 0.1% (v/v) formic acid) to 0.25 mL of the reaction system. The product concentrations and enantiomeric excess values (e.e.) were analyzed by HPLC, and the results are detailed in Table 6.In addition, CFE containing transaminase YP-955297 was co-warmed with phenylacetone and α-methylbenzylamine, which similarly produced α-ethylbenzylamine and acetophenone. The reaction was terminated and analyzed as above. After warming at 28°C for 24 h, 0.87 mmol/L of (R)-α-ethylbenzylamine was obtained with a high e.e. value. The above results indicate that transaminases XP-001209325 and YP-955297 are highly selective (R)-transaminases. In addition, the substrate profiles of XP-001209325 and the transaminases AAN21261 and ABN35871 differed significantly: XP-001209325 significantly catalyzed the generation of 4-phenyl-2-butylamine, whereas AAN21261 and ABN35871 did not (Table 6). In addition, XP-001209325 catalyzed the generation of enantiomerically enriched (R)-2-aminobutane from 2-butanone, whereas ABN35871 generated enantiomerically enriched (S)-2-aminobutane (Table 6).