A plasmid pET28aM01A82W
containing the M01A82W variant (R47L/A82W/F87V/L188Q/E267V) was used
as a template for the mutation.34 (link) Specifically,
under the action of Fast Mutagenesis System (TransGen Biotech, Beijing,
China), M01A82W was mutated into another BM3 variant M13 (R47L/L86I/F87V/L188Q),
thereby generating another plasmid pET28aM13.19 (link) Next, pET28aM13 was used as the template to yield diverse pET-28a
(+) derived plasmids containing M13 variants using Fast Mutagenesis
System. The primers used for the mutation are listed inTable S1 . The presence of the desired mutations
in M13 was confirmed by DNA sequencing.
E. coli strains Trans1-T1 and Transetta
(DE3) (TransGen Biotech, Beijing, China) served as hosts for recombinant
plasmid amplification and enzyme expression, respectively.
Genistein
was purchased from Yuanye (Shanghai yuanye Bio-Technology
Co., Ltd., Shanghai, China).
containing the M01A82W variant (R47L/A82W/F87V/L188Q/E267V) was used
as a template for the mutation.34 (link) Specifically,
under the action of Fast Mutagenesis System (TransGen Biotech, Beijing,
China), M01A82W was mutated into another BM3 variant M13 (R47L/L86I/F87V/L188Q),
thereby generating another plasmid pET28aM13.19 (link) Next, pET28aM13 was used as the template to yield diverse pET-28a
(+) derived plasmids containing M13 variants using Fast Mutagenesis
System. The primers used for the mutation are listed in
in M13 was confirmed by DNA sequencing.
E. coli strains Trans1-T1 and Transetta
(DE3) (TransGen Biotech, Beijing, China) served as hosts for recombinant
plasmid amplification and enzyme expression, respectively.
Genistein
was purchased from Yuanye (Shanghai yuanye Bio-Technology
Co., Ltd., Shanghai, China).
