1gk0

From Proteopedia

Revision as of 14:08, 5 November 2007

STRUCTURE-BASED PREDICTION OF MODIFICATIONS IN GLUTARYLAMIDASE TO ALLOW SINGLE-STEP ENZYMATIC PRODUCTION OF 7-AMINOCEPHALOSPORANIC ACID FROM CEPHALOSPORIN C

Overview

Glutarylamidase is an important enzyme employed in the commercial, production of 7-aminocephalosporanic acid, a starting compound in the, synthesis of cephalosporin antibiotics. 7-aminocephalosporanic acid is, obtained from cephalosporin C, a natural antibiotic, either chemically or, by a two-step enzymatic process utilizing the enzymes D-amino acid oxidase, and glutarylamidase. We have investigated possibilities for redesigning, glutarylamidase for the production of 7-aminocephalosporanic acid from, cephalosporin C in a single enzymatic step. These studies are based on the, structures of glutarylamidase, which we have solved with bound phosphate, and ethylene glycol to 2.5 A resolution and with bound glycerol to 2.4 A., The phosphate binds near the catalytic serine in a way that mimics the, hemiacetal that develops during catalysis, while the glycerol occupies the, side-chain binding pocket. Our structures show that the enzyme is not only, structurally similar to penicillin G acylase but also employs essentially, the same mechanism in which the alpha-amino group of the catalytic serine, acts as a base. A subtle difference is the presence of two catalytic, dyads, His B23/Glu B455 and His B23/Ser B1, that are not seen in, penicillin G acylase. In contrast to classical serine proteases, the, central histidine of these dyads interacts indirectly with the O(gamma), through a hydrogen bond relay network involving the alpha-amino group of, the serine and a bound water molecule. A plausible model of the, enzyme-substrate complex is proposed that leads to the prediction of, mutants of glutarylamidase that should enable the enzyme to deacylate, cephalosporin C into 7-aminocephalosporanic acid.

About this Structure

1GK0 is a Protein complex structure of sequences from Pseudomonas sp. with PO4 and EDO as ligands. Active as Penicillin amidase, with EC number 3.5.1.11 Structure known Active Site: POB. Full crystallographic information is available from OCA.

Reference

Structure-based prediction of modifications in glutarylamidase to allow single-step enzymatic production of 7-aminocephalosporanic acid from cephalosporin C., Fritz-Wolf K, Koller KP, Lange G, Liesum A, Sauber K, Schreuder H, Aretz W, Kabsch W, Protein Sci. 2002 Jan;11(1):92-103. PMID:11742126

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