The 2.0 Å crystal structure of cephalosporin acylase

被引:88
作者
Kim, Y [1 ]
Yoon, KH
Khang, Y
Turley, S
Hol, WGJ
机构
[1] Yeungnam Univ, Sch Chem Engn, Kyongsan 712749, South Korea
[2] Yeungnam Univ, Dept Appl Microbiol, Kyongsan 712749, South Korea
[3] Woosong Univ, Sch Food Biotechnol, Daejon 300100, South Korea
[4] Univ Washington, Dept Biol Struct, Seattle, WA 98195 USA
[5] Univ Washington, Dept Biochem, Seattle, WA 98195 USA
[6] Univ Washington, Biomol Struct Ctr, Seattle, WA 98195 USA
[7] Univ Washington, Howard Hughes Med Inst, Seattle, WA 98195 USA
关键词
cephalosporin acylase; cephalosporin C acylase; glutaryl-7ACA acylase; Ntn hydrolase;
D O I
10.1016/S0969-2126(00)00505-0
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Background: Semisynthetic cephalosporins are primarily synthesized from 7-aminocephalosporanic acid (7-ACA), which is usually obtained by chemical deacylation of cephalosporin C (CPC). The chemical production of 7-ACA includes, however, several expensive steps and requires thorough treatment of chemical wastes. Therefore, an enzymatic conversion of CPC to 7-ACA by cephalosporin acylase is of great interest. The biggest obstacle preventing this in industrial production is that cephalosporin acylase uses glutaryl-7ACA as a primary substrate and has low substrate specificity for CPC. Results: We have solved the first crystal structure of a cephalosporin acylase from Pseudomonas diminuta at 2.0 a resolution. The overall structure looks like a bowl with two "knobs" consisting of helix- and strand-rich regions, respectively. The active site is mostly formed by the distinctive structural motif of the N-terminal (Ntn) hydrolase superfamily. Superposition of the 61 residue active-site pocket onto that of penicillin G acylase shows an rmsd in C alpha positions of 1.38 Angstrom. This indicates structural similarity in the active site between these two enzymes, but their overall structures are elsewhere quite different. Conclusion: The substrate binding pocket of the P. diminuta cephalosporin acylase provides detailed insight into the ten key residues responsible for the specificity of the cephalosporin C side chain in four classes of cephalosporin acylases, and it thereby forms a basis for the design of an enzyme with an improved conversion rate of CPC to 7-ACA. The structure also provides structural evidence that four of the five different classes of cephalosporin acylases can be grouped into one family of the Ntn hydrolase superfamily.
引用
收藏
页码:1059 / 1068
页数:10
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