The invention relates to the technical field of
enzyme catalysis, and discloses a
transaminase mutant obtained by high-
throughput screening and application thereof. The parent
transaminase of the
transaminase mutant is derived from
Paracoccus denitrifics PD1222, and the parent transaminase has an
amino acid sequence as shown in SEQ ID NO. 1 (Sequence Identifier Number 1). In order to solve the problem of low catalytic activity of current transaminase, the
mutant is obtained by performing single-point or multi-point combined substitution on the 19th, 85th, 150th and / or 153rd amino acids in the sequence of SEQ ID NO.1. Preferably, the substitutions include, but are not limited to, F19W, F19L, F19M, F19P, F85M, F85L, Y150M, Y150F, V153S, V153C, V153A and the like. Particularly preferred mutants are combined mutants containing triple substitutions of V153S, F85M and F19W, and the catalytic activity of the mutants on a substrate 1-methoxy-2-
acetone is improved by about 12.5 times compared with that of
wild type transaminase. The relative
enzyme activity of the transaminase mutant provided by the invention is remarkably improved compared with that of a
wild type, and the transaminase mutant shows excellent tolerance to a high-concentration substrate.