The invention relates to the technical field of
gene engineering and
enzyme engineering, and particularly provides an alkaline
amylase mutant F178A with improved
enzyme activity as well as a preparation method and application of the alkaline
amylase mutant F178A. The
mutant is obtained by replacing 178th-site
phenylalanine (Phe) of an
amino acid sequence as shown in SEQ ID NO: 1 with
alanine (Ala), and the
amino acid sequence of the mutant is as shown in SEQ ID NO: 3. Compared with wild-type Ano
amylase, the mutant has the advantages that the
enzyme catalysis efficiency is obviously improved, the
specific enzyme activity to soluble
starch, corn raw
starch, wheat raw
starch and cassava raw starch is higher than that of the original enzyme, and meanwhile, the excellent properties such as the optimum temperature (70 DEG C), the optimum pH (8.0) and the
thermal stability of the mutant are completely reserved.
Molecular simulation shows that
mutation increases the volume of an active pocket and is beneficial to substrate binding and product release. The mutant provided by the invention has wide application prospects in the industrial fields of
textile desizing, starch saccharification,
biofuel production and the like.