Cloning, characterization and molecular docking of a highly thermostable β-1,4-glucosidase from Thermotoga petrophila
| Autor: | Naeem Rashid, Mahmood Ali Khan, Zahid Hussain, Sumra Afzal, Ikram ul Haq, Bushra Muneer, Muhammad Mohsin Javed, Ishtiaq Ahmad, Sana Majeed |
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| Rok vydání: | 2012 |
| Předmět: |
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Molecular Bioengineering Molecular Dynamics Simulation Biology medicine.disease_cause Polymerase Chain Reaction Applied Microbiology and Biotechnology Substrate Specificity Bacteria Anaerobic Glucosides Escherichia coli medicine Cloning Molecular Gene Thermotoga petrophila chemistry.chemical_classification Cloning Glucan 1 4-beta-Glucosidase Hydrolysis General Medicine Recombinant Proteins Molecular Weight Kinetics genomic DNA Enzyme chemistry Biochemistry Docking (molecular) Product inhibition Electrophoresis Polyacrylamide Gel Biotechnology |
| Zdroj: | Biotechnology Letters. 34:1703-1709 |
| ISSN: | 1573-6776 0141-5492 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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