Structural characterization of the bglH gene encoding a beta-glucosidase-like enzyme in an endophytic Bacillus pumilus strain

Andréa C. Bogas, Maria Angelica E. Watanabe, Aneli Barbosa, Laurival A. Vilas-Boas, Ana C. Bonatto, Robert Dekker, Emanuel M. Souza, Maria Helena P. Fungaro
2007 Genetics and Molecular Biology  
A beta-glucosidase-like enzyme-encoding gene (bglH) of an endophytic Bacillus pumilus strain (CL16) was cloned using a shotgun genomic library constructed in Escherichia coli. The nucleotide sequence of the entire cloned fragment (2484 bp) was determined and characterized. An incomplete open reading frame (ORF) of 534 bp (ORF1) designated bglP and a complete ORF of 1419 bp (ORF2) designated bglH, located in the fragment, are organized in an operon. The protein deduced from 1419 bp (ORF2) had
more » ... 19 bp (ORF2) had 472 amino acid residues without a characteristic signal peptide sequence, suggesting that the enzyme is localized in the cytoplasm. The amino acid sequence deduced from bglH gene had high similarity with β-glucosidases from the glycosyl hydrolase family 1. Over-expression of the B. pumilus bglH gene in E. coli showed a 54 kDa protein whose identity was confirmed by mass spectrometry (MALDI-TOF). Using the degenerated primers DEG1F (5'-ATRACC TACTgNAARTTRgg-3') and DEG1R (5'gCRAANCCY AgHTARACggT-3') designed based on the amino acid regions conserved amongst β-glucosidases reported for
doi:10.1590/s1415-47572007000100018 fatcat:xcz7sggqvfe27cd76mesywa5n4