Cloning and characterization of a bacterial cell-bound type B carboxylesterase from Bacillus sp BP-7

A clone producing halos on tributyrin plates was isolated from a genomic li brary of Bacillus sp, BP-7. The insert contained an open reading frame that coded for a protein of 487 amino acids with homology to carboxylesterases. The cloned enzyme showed clear preference for esters of short-chain fatty acids, being classified as an esterase. Maximum activity was found at 45 de greesC and pH 7.5. The enzyme displayed stability in the pH range from 6 to 9.5, and at temperatures from 4 degrees to 45 degreesC. Zymogram analysis of the protein revealed a molecular mass of 53 kDa and a pi of 5.1. The enz yme showed homology to members of the bacterial subclass of type B carboxyl esterases, a set of proteins potentially useful for biotechnological applic ations.