E. Gelhaye et al., EFFECTS OF CELLOBIOSE ON CELLULOSE COLONIZATION BY A MESOPHILIC, CELLULOLYTIC CLOSTRIDIUM (STRAIN-C401), Journal of General Microbiology, 139, 1993, pp. 2819-2824
When cultured on a mixture of cellobiose and cellulose, Clostridium C4
01 did not initially attach to cellulose but remained in the liquid ph
ase. After cellobiose exhaustion, bacterial cells grew in association
with the insoluble cellulose. Carboxymethylcellulase (CMCase) producti
on on Avicel cellulose was four- to fivefold greater than on cellobios
e, and cellulose-grown cells adhered to filter paper with an initial a
dhesion rate about four- to fivefold greater than did cellobiose-grown
cells. Using tritiated thymidine incorporation as a measure of growth
, it appeared that transfer of strain C401 from cellobiose to cellulos
e required an adaptation phase. An extracellular cellulase complex was
isolated by affinity chromatography. This enzyme system is a multicom
ponent aggregate (molecular mass above 5 MDa), and yielded two major p
olypeptide bands by SDS-PAGE having molecular masses of 130 and 70 kDa
. Cellobiose strongly inhibited Avicelase activity and slightly inhibi
ted p-nitrophenylcellobiose hydrolysis (pNPCbase), but had no effect o
n the CMCase activity of the cellulase complex. In addition, polyclona
l antibodies, raised against the purified 130 kDa protein inhibited Av
icelase activity, but not CMCase and pNPCbase activities.