TY - JOUR
T1 - Structure of cephalosporin acylase in complex with glutaryl-7-aminocephalosporanic acid and glutarate
T2 - Insight into the basis of its substrate specificity
AU - Kim, Youngsoo
AU - Hol, Wim G.J.
N1 - Funding Information:
We thank Ethan Merritt, Stephen Suresh and Craig Behnke for their helpful discussions, Jungwoo Choe, Stewart Turley and Mic Feese for collecting data, and the SBC-CAT staff, in particular Randy Alkire, Stephan Ginell and Rongguang Zhang for their technical assistance on the APS SBC-CAT beamline. Use of the Argonne National Laboratory Structural Biology Center beamlines at the APS was supported by the US Department of Energy Office of Energy Research, under contract No. W-31-109-ENG-38. We also thank Francis Athappilly, Irwin Hirsh, and Claudia Roach of the Biomolecular Structure Center for maintaining our computer facilities and helping with the protein expression, purification and crystallization. W.G.J.H. acknowledges a major equipment grant from the Murdock Charitable Trust to the Biomolecular Structure Center. This study was supported by a Grant of the Korea Health 21 R&D Project, Ministry of Health and Welfare, Republic of Korea (01-PJ1-PG3-20900-0009) to Y.K.
PY - 2001
Y1 - 2001
N2 - Background: Semisynthetic cephalosporins are primarily synthesized from 7-aminocephalosporanic acid (7-ACA), which is obtained by environmentally toxic chemical deacylation of cephalosporin C (CPC). Thus, the enzymatic conversion of CPC to 7-ACA by cephalosporin acylase (CA) would be of great interest. However, CAs use glutaryl-7-ACA (GL-7-ACA) as a primary substrate and the enzyme has low turnover rates for CPC. Results: The binary complex structures of CA with GL-7-ACA and glutarate (the side-chain of GL-7-ACA) show extensive interactions between the glutaryl moiety of GL-7-ACA and the seven residues that form the side-chain pocket. These interactions explain why the D-α-aminoadipyl side-chain of CPC yields a poorer substrate than GL-7-ACA. Conclusions: This understanding of the nature of substrate specificity may be useful in the design of an enzyme with an improved performance for the conversion of CPC to 7-ACA. Additionally, the catalytic mechanism of the deacylation reaction was revealed by the ligand bound structures.
AB - Background: Semisynthetic cephalosporins are primarily synthesized from 7-aminocephalosporanic acid (7-ACA), which is obtained by environmentally toxic chemical deacylation of cephalosporin C (CPC). Thus, the enzymatic conversion of CPC to 7-ACA by cephalosporin acylase (CA) would be of great interest. However, CAs use glutaryl-7-ACA (GL-7-ACA) as a primary substrate and the enzyme has low turnover rates for CPC. Results: The binary complex structures of CA with GL-7-ACA and glutarate (the side-chain of GL-7-ACA) show extensive interactions between the glutaryl moiety of GL-7-ACA and the seven residues that form the side-chain pocket. These interactions explain why the D-α-aminoadipyl side-chain of CPC yields a poorer substrate than GL-7-ACA. Conclusions: This understanding of the nature of substrate specificity may be useful in the design of an enzyme with an improved performance for the conversion of CPC to 7-ACA. Additionally, the catalytic mechanism of the deacylation reaction was revealed by the ligand bound structures.
KW - Cephalosporin acylase
KW - Cephalosporin antibiotic
KW - Glutaryl-7-aminocephalosporanic acid
KW - Substrate specificity
UR - http://www.scopus.com/inward/record.url?scp=0035542860&partnerID=8YFLogxK
U2 - 10.1016/S1074-5521(01)00092-8
DO - 10.1016/S1074-5521(01)00092-8
M3 - Article
C2 - 11755403
AN - SCOPUS:0035542860
VL - 8
SP - 1253
EP - 1264
JO - Chemistry and Biology
JF - Chemistry and Biology
SN - 1074-5521
IS - 12
ER -
