Cloning and characterization of cel8A gene from Rhizobium leguminosarum bv. trifolii 1536*

Abstract

Aims: To isolate the cellulase gene from Rhizobium leguminosarum bv. trifolii 1536. Methods and Results: By the shot-gun method a clone (cel8A) harbouring 3AE1 kb genomic DNA fragment from R. leguminosarum bv. trifolii 1536 was obtained. The cel8A gene coded 348 amino acids and it belongs to the glycosyl hydrolase family 8. The molecular mass of Cel8A protein induced from Escherichia coli DH5a, appeared to be 35 kDa. The optimum pH and optimum temperature was 7AE0, and about 30°C for its enzymatic activity respectively. Conclusions: R. leguminosarum bv. trifolii 1536 had cel8A gene having an open reading frame of 1047 bp coded for the activity of hydrolyzation of carboxymethyl cellulose. Significance and Impact of the Study: The production of celluloytic enzyme by R. leguminosarum bv. trifolii was confirmed, which would play specific roles in rhizobia. Future study should focus on its role in the infection and nodulation phenomena.

Electron microscopy analysis of carboxymethylcellulase in rhizobia

“…trifolii 1536 was obtained. The cel8A gene encodes a glycosyl hydrolase family 8 member of 348 amino acids that exhibits a molecular mass of 35 kDa when induced from Escherichia coli DH5a (An et al, 2004).…”