TY - JOUR
T1 - Structural insights into the substrate recognition properties of β-glucosidase
AU - Nam, Ki Hyun
AU - Sung, Min Woo
AU - Hwang, Kwang Yeon
N1 - Funding Information:
We thank Dr. H.S. Lee, K.J. Kim, and K.H. Kim for their kind assistance with the data collection using beamlines 4A and 6C of the Pohang Light Source, Korea. This study was supported by the Korea Science and Engineering Foundation. [R01-2007-000-20072-0 (2009)]. K.H. Nam is supported by the BK21 program.
PY - 2010/1/1
Y1 - 2010/1/1
N2 - β-Glucosidase enzymes (EC 3.2.1-3.2.3) hydrolyze sugars and are implicated in a wide spectrum of biological processes. Recently, we reported that β-glucosidase has varied kinetic parameters for the natural and synthetic substrates [K.H Nam, S.J. Kim, M.Y. Kim, J.H. Kim, T.S. Yeo, C.M. Lee, H.K Jun, K.Y. Hwang. Crystal structure of engineered beta-glucosidase from a soil metagenome, Proteins 73 (2008) 788-793]. However, an understanding of the kinetic values of β-glucosidase has not yet enabled the elucidation of its molecular function. Here, we report the X-ray crystal structure of β-glucosidase with a glucose and cellobiose fragment from uncultured soil metagenome. From the various crystals, we obtained the pre-reaction (native), intermediate (disaccharide cleavage) and post-reaction (glucose binding) states of the active site pocket. These structures provide snapshots of the catalytic processing of β-glucosidase. In addition, the intermediate state of the crystal structure provides insight into the substrate specificity of β-glucosidase. These structural studies will facilitate elucidation of the architectural mechanism responsible for the substrate recognition of β-glucosidase.
AB - β-Glucosidase enzymes (EC 3.2.1-3.2.3) hydrolyze sugars and are implicated in a wide spectrum of biological processes. Recently, we reported that β-glucosidase has varied kinetic parameters for the natural and synthetic substrates [K.H Nam, S.J. Kim, M.Y. Kim, J.H. Kim, T.S. Yeo, C.M. Lee, H.K Jun, K.Y. Hwang. Crystal structure of engineered beta-glucosidase from a soil metagenome, Proteins 73 (2008) 788-793]. However, an understanding of the kinetic values of β-glucosidase has not yet enabled the elucidation of its molecular function. Here, we report the X-ray crystal structure of β-glucosidase with a glucose and cellobiose fragment from uncultured soil metagenome. From the various crystals, we obtained the pre-reaction (native), intermediate (disaccharide cleavage) and post-reaction (glucose binding) states of the active site pocket. These structures provide snapshots of the catalytic processing of β-glucosidase. In addition, the intermediate state of the crystal structure provides insight into the substrate specificity of β-glucosidase. These structural studies will facilitate elucidation of the architectural mechanism responsible for the substrate recognition of β-glucosidase.
KW - Catalysis processing
KW - Intermediate state
KW - Steric hindrance
KW - Substrate specificity
KW - β-Glucosidase
UR - http://www.scopus.com/inward/record.url?scp=72949100309&partnerID=8YFLogxK
U2 - 10.1016/j.bbrc.2009.12.038
DO - 10.1016/j.bbrc.2009.12.038
M3 - Article
C2 - 20005197
AN - SCOPUS:72949100309
VL - 391
SP - 1131
EP - 1135
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 1
ER -