Cloning, characterization and molecular docking of a highly thermostable β-1,4-glucosidase from Thermotoga petrophila.
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.
Institute of Industrial Biotechnology, GC University, Lahore 54000, Pakistan. chemistrymak@yahoo.com
Ikram Ul
IU
Haq
Mahmood Ali
MA
Khan
Bushra
B
Muneer
Zahid
Z
Hussain
Sumra
S
Afzal
Sana
S
Majeed
Naeem
N
Rashid
Muhammad Mohsin
MM
Javed
Ishtiaq
I
Ahmad
D001421:Bacteria, Anaerobic / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D044643:Glucan 1,4-beta-Glucosidase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D005960:Glucosides / Q000378:metabolism; D006868:Hydrolysis; D007700:Kinetics; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D008970:Molecular Weight; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity
D005960:Glucosides; D011994:Recombinant Proteins; C025193:4-nitrophenyl beta-D-glucoside; D044643:Glucan 1,4-beta-Glucosidase