Table 1.

Screening of extracellular PCX cellulase activity of knockout and original S. cerevisiae BY4741 strains by 4-MUC assay (recombinant/original ratio)

Gene null mutant	ORF namea	96-well screening 4-MUC assay	Condensed sample 4-MUC assay (o/n OD600)
Protein-trafficking genes:
chc1∆::KanMX	YGL206C	2.5	2.4 (2.8)
clc1∆::KanMX	YGR167W	3.8	2.6 (3.1)
nyv1∆::KanMX	YLR093Cf	1.5	1.7 (5.7)
pep12∆::KanMX	YOR036W	1.9	2.6 (2.9)
vam3∆::KanMX	YOR106W	1.6	NA
vps1∆::KanMX	YKR001C	1.2	NA
vps15∆::KanMX	YBR097W	1.9	1.3 (4.2)
vps21∆::KanMX	YOR089Cf	3.3	6.5 (6.1)
vps33∆::KanMX	YLR396C	1.5	0.9 (5.5)
ypt7∆::KanMX	YML001Wb	1.7	1.0 (5.1)
Glycosylation-related genes:
aga2∆::KanMX	YGL032C	0.9	NA
alg3∆::KanMX	YBL082Cc	0.5	NA
alg5∆::KanMX	YPL227Cc	0.7	NA
alg6∆::KanMX	YOR002Wc	0.8	NA
alg8∆::KanMX	YOR067Cc	1.5	NA
alg9∆::KanMX	YNL219Cc	1.5	NA
alg12∆::KanMX	YNR030Wc	1.7	NA
anp1∆::KanMX	YEL036Cc	1.8	NA
csg2∆::KanMX	YBR036C	1.0	NA
cwh41∆::KanMX	YGL027Cc	0.1	NA
cwh43∆::KanMX	YCR017C	1.3	NA
die2∆::KanMX	YGR227Wc	1.5	NA
gda1∆::KanMX	YEL042W	1.6	NA
gnt1∆::KanMX	YOR320C	1.2	NA
gon7∆::KanMX	YJL184W	0.1	NA
gtb1∆::KanMX	YDR221Wd,e	2.7	0.0 (12.6)
hoc1∆::KanMX	YJR075W	0.9	NA
hor7∆::KanMX	YMR251W-A	1.4	NA
ktr1∆::KanMX	YOR099Wd	0.8	NA
ktr2∆::KanMX	YKR061Wd,f	0.9	NA
ktr3∆::KanMX	YBR205Wd	1.0	NA
ktr4∆::KanMX	YBR199Wd	1.0	NA
ktr5∆::KanMX	YNL029Cc	1.0	NA
ktr6∆::KanMX	YPL053Cd	1.3	NA
ktr7∆::KanMX	YIL085Cd	1.0	NA
mnl1∆::KanMX	YHR204Wd	1.2	NA
mnn2∆::KanMX	YBR015Cc	1.3	0.8 (3.8)
mnn5∆::KanMX	YJL186Wf	1.9	3.0 (5.6)
mnn9∆::KanMX	YPL050Cc,f	2.2	3.8 (5.5)
mnn10∆::KanMX	YDR245W	2.5	6.0 (3.8)
mnn11∆::KanMX	YJL183Wc	2.0	4.5 (3.7)
mns1∆::KanMX	YJR131Wd	0.5	NA
mnt2∆::KanMX	YGL257Cd	1.2	NA
mnt3∆::KanMX	YIL014Wd,e	2.5	1.7 (5.2)
mrl1∆::KanMX	YPR079Wf	1.6	NA
ost3∆::KanMX	YOR085Wc	1.1	NA
ost4∆::KanMX	YDL232Wc,f	1.6	NA
ost5∆::KanMX	YGL226C-Ac	0.9	NA
ost6∆::KanMX	YML019Wc,f	1.8	NA
pmt1∆::KanMX	YDL095Wc	3.1	3.2 (4.7)
pmt2∆::KanMX	YAL023Cc	5.3	1.8 (4.7)
pmt3∆::KanMX	YOR321Wc	1.1	1.0 (4.4)
pmt5∆::KanMX	YDL093Wc	2.1	4.3 (6.3)
pmt6∆::KanMX	YGR199Wc	0.8	NA
ynd1∆::KanMX	YER005Wf	1.1	NA
yur1∆::KanMX	YJL139Cf	2.0	2.8(4.5)
yjr061w∆::KanMX	YJR061W	1.0	NA

^aYGL038C, YLR240W, YGL095C knockout strains failed to be transformed with PCX.

^bEnhancement of Ctcel8A endocellulase activity [52].

Enhancement^c and decrease^d of Cel8Aenz endocellulase activity [48].

^eDetectable NpaBGS beta-glucosidase activity as screened by pNPG assay.

^fThe null mutant could grow in SC-URA medium with 1% cellobiose when the NpaBGS beta-glucosidase gene is functional inside.

The 4-MUC activity ratio is the average PCX activity of the three highest expressing recombinants (single-gene-KO strain) to the average original PCX activity (BY4741 strain).