American Journal of Microbiological Research
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American Journal of Microbiological Research. 2018, 6(4), 173-180
DOI: 10.12691/ajmr-6-4-5
Open AccessArticle

Characterization of Thermophilic β-Glucosidase of Rhizospheric Bacterial Strain (LSKB15) Isolated from Cholistan Desert, Pakistan

Amer Ahmed1, , Faiz ul Hassan Nasim2, Kashfa Batool2, Aasia Bibi3 and Sarwat Rafi2

1Department of Biochemistry and Biotechnology, Faculty of Science, The Islamia University of Bahawalpur, Bahawalpur, Pakistan

2Department of Chemistry, Biochemistry Section, Faculty of Science, The Islamia University of Bahawalpur, Bahawalpur, Pakistan

3Department of Biochemical Sciences, School of Molecular Biology and Medicine, La Sapienza University of Rome, Rome, Italy

Pub. Date: November 09, 2018

Cite this paper:
Amer Ahmed, Faiz ul Hassan Nasim, Kashfa Batool, Aasia Bibi and Sarwat Rafi. Characterization of Thermophilic β-Glucosidase of Rhizospheric Bacterial Strain (LSKB15) Isolated from Cholistan Desert, Pakistan. American Journal of Microbiological Research. 2018; 6(4):173-180. doi: 10.12691/ajmr-6-4-5

Abstract

Fifty thermophilic bacterial strains isolated from rhizospheric soil of Cholistan desert, Pakistan, and designated as LSKB01-LSKB50 were screened for β-glucosidase gene (bgl) belonging to glycoside hydrolase family 1 (GH 1) using PCR technique. Subsequently, the same strains were screened for extracellular β-glucosidase production using esculin as substrate. All fifty strains were shown to be amplified for conserved region of bgl gene and to secrete extracellular β-glucosidase. One strain (LSKB15) secreted relative high amount of this enzyme as indicating by size of ferric-esculetin precipitate. This strain was further cultivated on cellulose containing media and β-glucosidase was purified by ammonium sulfate, dialysis and gel filtration chromatography. The purified enzyme showed an optimal temperature of 60°C and an optimal pH of 7. It also showed excellent temperature and pH stability retaining > 90% activity after incubation for 2 h at pH 5-8 and 40-60°C. Finally, the purified enzyme was run on Native-PAGE and subsequently incubated in phosphate buffer containing 5 mM of 4-methylumbelliferyl-β-D-glucoside (4-MUG) for 15 min at 50°C and visualized by UV light as white band. We concluded that thermophilic LSKB15 β- glucosidase may work with other cellulase to degrade available cellulose synthesized by plant and the properties exhibited by it such as high temperature and pH stability pointed out its potential industrial importance.

Keywords:
Screening cellulose cellulase β-glucosidase thermophile PCR biofuel

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