Cloning, characterization and molecular docking of a highly thermostable β-1,4-glucosidase from Thermotoga petrophila.

Ikram Ul Haq, Mahmood Ali Khan, Bushra Muneer, Zahid Hussain, Sumra Afzal, Sana Majeed, Naeem Rashid, Muhammad Mohsin Javed, Ishtiaq Ahmad

Institute of Industrial Biotechnology, GC University, Lahore 54000, Pakistan. chemistrymak@yahoo.com

Biotechnology letters 2012 Sep

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A genomic DNA fragment, encoding a thermotolerant β-glucosidase, of the obligate anaerobe Thermotoga petrophila RKU-1 was cloned after PCR amplification into Escherichia coli strain BL21 CodonPlus. The purified cloned enzyme was a monomeric, 51.5 kDa protein (by SDS-PAGE) encoded by 1.341 kb gene. The estimated K (m) and V (max) values against p-nitrophenyl-β-D-glucopyranoside were 2.8 mM and 42.7 mmol min(-1) mg(-1), respectively. The enzyme was also active against other p-nitrophenyl substrates. Possible catalytic sites involved in hydrolyzing different p-nitrophenyl substrates are proposed based on docking studies of enzyme with its substrates. Because of its unique characters, this enzyme is a potential candidate for industrial applications.

Citation

Ikram Ul Haq, Mahmood Ali Khan, Bushra Muneer, Zahid Hussain, Sumra Afzal, Sana Majeed, Naeem Rashid, Muhammad Mohsin Javed, Ishtiaq Ahmad. Cloning, characterization and molecular docking of a highly thermostable β-1,4-glucosidase from Thermotoga petrophila. Biotechnology letters. 2012 Sep;34(9):1703-9