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

Abstract

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 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).

Production of bacterial amylases and cellulases using sweet potato (Ipomoea batatas. (L.) Lam.) peels

“…The prevalence of B. pumulis and B. subtilis isolated in this work conforms to the findings of Lorena et al (2001) and Madigan et al (2005) which states that these two organisms are natural inhabitant of soil. In this study, B. pumilus produced the highest concentration of amylase (5.4 U/mL) at 55°C and pH 6 which was also reported by Andrea et al (2007) which states that B. pumilus produced amylase between the pH of 5.8 and 7.5 and at a temperature of 55°C. Effect of temperature on amylase production was observed by varying growth temperature of isolates and optimum temperature was found to be 55°C.…”

Brevibacillus Thermoruber 9X-GLC, Bacteria Isolated from Hot Compost, Producer of a Beta-Glucosidase Resistant to Glucose Inhibition

“…Enzymatic degradation of cellulose is an important step in bioethanol production from plant biomass, it requires synergistic action of multiple enzymes, mainly endo-β-1,4glucanase, cellobiohydrolase (exo-glucanase) and β-glucosidase (Fujita et al, 2004). Beta-glucosidases are present in bacteria, fungi and plants and show hydrolytic activity on cellobiose and aromatic compounds such as arbutin and silicin present in some plant tissues (Tajima et al, 2001;Spiridonov and Wilson, 2001;Park et al, 2002;Bogas et al, 2007). Enzymatic systems lacking β-glucosidase or those producing it in small amounts show incomplete saccharification of plant cellulose and usually show product inhibition (Gusakov and Sinitsyn, 1992).…”