Regulation of Expression of Cellulosomal Cellulase and Hemicellulase Genes in Clostridium cellulovorans

Abstract

ABSTRACT The regulation of expression of the genes encoding the cellulases and hemicellulases of Clostridium cellulovorans was studied at the mRNA level with cells grown under various culture conditions. A basic pattern of gene expression and of relative expression levels was obtained from cells grown in media containing poly-, di- or monomeric sugars. The cellulase ( cbpA and engE ) and hemicellulase ( xynA ) genes were coordinately expressed in medium containing cellobiose or cellulose. Growth in the presence of cellulose, xylan, and pectin gave rise to abundant expression of most genes ( cbpA - exgS , engH , hbpA , manA , engM , engE , xynA , and/or pelA ) studied. Moderate expression of cbpA , engH , manA , engE , and xynA was observed when cellobiose or fructose was used as the carbon source. Low levels of mRNA from cbpA , manA , engE , and xynA were observed with cells grown in lactose, mannose, and locust bean gum, and very little or no expression of cbpA , engH , manA , engE , and xynA was detected in glucose-, galactose-, maltose-, and sucrose-grown cells. The cbpA - exgS and engE genes were most frequently expressed under all conditions studied, whereas expression of xynA and pelA was more specifically induced at higher levels in xylan- or pectin-containing medium, respectively. Expression of the genes ( cbpA , hbpA , manA , engM , and engE ) was not observed in the presence of most soluble di- or monosaccharides such as glucose. These results support the hypotheses that there is coordinate expression of some cellulases and hemicellulases, that a catabolite repression type of mechanism regulates cellulase expression in rapidly growing cells, and that the presence of hemicelluloses has an effect on cellulose utilization by the cell.