title: Characterization of a recombinant aryl β-glucosidase from Neosartorya fischeri NRRL181.
abstract: An isolated gene from Neosartorya fischeri NRRL181 encoding a β-glucosidase (BGL) was cloned, and its nucleotide sequence was determined. DNA sequence analysis revealed an open reading frame of 1,467 bp, capable of encoding a polypeptide of 488 amino acid residues. The gene was over-expressed in Escherichia coli, and the protein was purified using nickel-nitrilotriacetic acid chromatography. The purified recombinant BGL showed a high level of catalytic activity, with V                         (max) of 886 μmol min(-1) mg-protein(-1) and a K                         (m) of 68 mM for p-nitrophenyl-β-D: -glucopyranoside (pNPG). The optimal temperature for enzyme activity was about 40°C, and the optimal pH was about 6.0. A homology model of N. fischeri BGL1 was constructed based on the X-ray crystal structure of Phanerochaete chrysosporium BGLA. Molecular dynamics simulation studies of the enzyme with the pNPG and cellobiose shed light on the unique substrate specificity of N. fischeri BGL1 only towards pNPG.
authors: Department of Chemical Engineering, Konkuk University, Seoul, South Korea.
authors: Dayanand
authors: D
authors: Kalyani
authors: Kyoung-Mi
authors: KM
authors: Lee
authors: Manish Kumar
authors: MK
authors: Tiwari
authors: Priyadharshini
authors: P
authors: Ramachandran
authors: Hoon
authors: H
authors: Kim
authors: In-Won
authors: IW
authors: Kim
authors: Marimuthu
authors: M
authors: Jeya
authors: Jung-Kul
authors: JK
authors: Lee
mesh_terms: D000595:Amino Acid Sequence; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D055326:Neosartorya / Q000201:enzymology* / Q000235:genetics*; D016366:Open Reading Frames; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013696:Temperature; D001617:beta-Glucosidase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*
chemical_list: D004271:DNA, Fungal; D011994:Recombinant Proteins; D001617:beta-Glucosidase