Improvements of the productivity and saccharification efficiency of the cellulolytic β-glucosidase D2-BGL in Pichia pastoris via directed evolution

Table 1 Enzyme activity in culture supernatant and specific activity for P. pastoris-expressed wild-type and mutant D2-BGL

Pichia pastoris strain	Mutation	Activity^{a, b} (U/mL)	Relative activity^c (%)	Specific activity^b (U/mg)
WT D2		2.31 ± 0.20	100 ± 9	213 ± 8
Mut A	D358G + S496P	2.24 ± 0.04	97 ± 2	218 ± 1
D358G	D358G	1.77 ± 0.21	77 ± 9	164 ± 10
S496P	S496P	1.94 ± 0.12	84 ± 5	182 ± 8
Mut B	D224G + S592T	4.29 ± 0.42	186 ± 18	229 ± 19
D224G	D224G	4.08 ± 0.36	177 ± 16	202 ± 19
S592T	S592T	1.72 ± 0.51	75 ± 22	195 ± 13
Mut C	F256M + Y260D	4.09 ± 0.52	177 ± 23	253 ± 29
F256M	F256M	3.01 ± 0.59	130 ± 25	301 ± 26
Y260D	Y260D	1.92 ± 0.38	83 ± 17	189 ± 11
D224G F256M	D224G + F256M	5.70 ± 0.26	247 ± 11	236 ± 14
D224G Y260D	D224G + Y260D	4.63 ± 0.70	200 ± 30	192 ± 4
Mut M	D224G + F256M + Y260D	7.5 ± 0.75	325 ± 33	261 ± 23

ISSN: 2731-3654