USE OF A NEW MEMBRANE-REACTOR SACCHARIFICATION ASSAY TO EVALUATE THE PERFORMANCE OF CELLULASES UNDER SIMULATED SSF CONDITIONS - EFFECT ON ENZYME QUALITY OF GROWING TRICHODERMA-REESEI IN THE PRESENCE OF TARGETED LIGNOCELLULOSIC SUBSTRATE
Jo. Baker et al., USE OF A NEW MEMBRANE-REACTOR SACCHARIFICATION ASSAY TO EVALUATE THE PERFORMANCE OF CELLULASES UNDER SIMULATED SSF CONDITIONS - EFFECT ON ENZYME QUALITY OF GROWING TRICHODERMA-REESEI IN THE PRESENCE OF TARGETED LIGNOCELLULOSIC SUBSTRATE, Applied biochemistry and biotechnology, 63-5, 1997, pp. 585-595
A new saccharification assay has been devised; in which a continuously
buffer-swept membrane reactor is used to remove the solubilized sacch
arification products, thus allowing high extents of substrate conversi
on without significant inhibitory effects from the buildup of either c
ellobiose or glucose. This diafiltration saccharification assay (DSA)
can, therefore, be used to obtain direct measurements of the performan
ce of combinations of cellulase and substrate under simulated SSF cond
itions, without the saccharification results being complicated by fact
ors that may influence the subsequent fermentation step. This assay ha
s been used to compare the effectiveness of commercial and special in-
house produced Trichoderma reesei cellulase preparations in the saccha
rification of a standardized microcrystalline (Sigmacell) substrate an
d a dilute-acid pretreated lignocellulosic substrate. Initial results
strongly suggest that enzyme preparations produced in the presence of
the targeted lignocellulosic substrate will saccharify that substrate
more effectively. These results call into question the widespread use
of the ''filter paper assay'' as a reliable predictor of enzyme perfor
mance in the extensive hydrolysis of substrates that are quite differe
nt from filter paper in both physical properties and chemical composit
ion.