Tyr320 is a molecular determinant of the catalytic activity of β-glucosidase from Neosartorya fischeri
Research output: Contribution to journal › Journal article › Research › peer-review
Standard
Tyr320 is a molecular determinant of the catalytic activity of β-glucosidase from Neosartorya fischeri. / Shanmugam, Ramasamy; Kim, In Won; Tiwari, Manish K.; Gao, Hui; Mardina, Primata; Das, Devashish; Kumar, Anurag; Jeya, Marimuthu; Kim, Sang Yong; Kim, Young Sin; Lee, Jung Kul.
In: International Journal of Biological Macromolecules, Vol. 151, 15.05.2020, p. 609-617.Research output: Contribution to journal › Journal article › Research › peer-review
Harvard
APA
Vancouver
Author
Bibtex
}
RIS
TY - JOUR
T1 - Tyr320 is a molecular determinant of the catalytic activity of β-glucosidase from Neosartorya fischeri
AU - Shanmugam, Ramasamy
AU - Kim, In Won
AU - Tiwari, Manish K.
AU - Gao, Hui
AU - Mardina, Primata
AU - Das, Devashish
AU - Kumar, Anurag
AU - Jeya, Marimuthu
AU - Kim, Sang Yong
AU - Kim, Young Sin
AU - Lee, Jung Kul
PY - 2020/5/15
Y1 - 2020/5/15
N2 - β-Glucosidases (BGL) are key members of the cellulase enzyme complex that determine efficiency of lignocellulosic biomass degradation, which have shown great functional importance to many biotechnological systems. A previous reported BGL from Neosartorya fischeri (NfBGL) showed much higher activity than other BGLs. Screening the important residues based on sequence alignment, analyzing a homology model, and subsequent alteration of individually screened residues by site-directed mutagenesis were carried out to investigate the molecular determinants of the enzyme's high catalytic efficiency. Tyr320, located in the wild-type NfBGL substrate-binding pocket was identified as crucial to the catalytic function of NfBGL. The replacement of Tyr320 with aromatic amino acids did not significantly alter the catalytic efficiency towards p-nitrophenyl β-D-glucopyranoside (pNPG). However, mutants with charged and hydrophilic amino acids showed almost no activity towards pNPG. Computational studies suggested that an aromatic acid is required at position 320 in NfBGL to stabilize the enzyme-substrate complex formation. This knowledge on the mechanism of action of the molecular determinants can also help rational protein engineering of BGLs.
AB - β-Glucosidases (BGL) are key members of the cellulase enzyme complex that determine efficiency of lignocellulosic biomass degradation, which have shown great functional importance to many biotechnological systems. A previous reported BGL from Neosartorya fischeri (NfBGL) showed much higher activity than other BGLs. Screening the important residues based on sequence alignment, analyzing a homology model, and subsequent alteration of individually screened residues by site-directed mutagenesis were carried out to investigate the molecular determinants of the enzyme's high catalytic efficiency. Tyr320, located in the wild-type NfBGL substrate-binding pocket was identified as crucial to the catalytic function of NfBGL. The replacement of Tyr320 with aromatic amino acids did not significantly alter the catalytic efficiency towards p-nitrophenyl β-D-glucopyranoside (pNPG). However, mutants with charged and hydrophilic amino acids showed almost no activity towards pNPG. Computational studies suggested that an aromatic acid is required at position 320 in NfBGL to stabilize the enzyme-substrate complex formation. This knowledge on the mechanism of action of the molecular determinants can also help rational protein engineering of BGLs.
KW - Catalytic efficiency
KW - MD simulation
KW - Substrate affinity
KW - β-Glucosidase
KW - π-Sigma interaction
U2 - 10.1016/j.ijbiomac.2020.02.117
DO - 10.1016/j.ijbiomac.2020.02.117
M3 - Journal article
C2 - 32061700
AN - SCOPUS:85080053751
VL - 151
SP - 609
EP - 617
JO - International Journal of Biological Macromolecules
JF - International Journal of Biological Macromolecules
SN - 0141-8130
ER -
ID: 237412050