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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
1
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pubmed:dateCreated |
2001-12-19
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pubmed:databankReference |
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pubmed:abstractText |
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.
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/11742126-11051090,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11742126-11080627,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11742126-1390755,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11742126-13955341,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11742126-1749775,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11742126-2993240,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11742126-4846962,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11742126-7477383,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11742126-7607251,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11742126-7816145,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11742126-9095194,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11742126-9721298
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Jan
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pubmed:issn |
0961-8368
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
11
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
92-103
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:11742126-Amidohydrolases,
pubmed-meshheading:11742126-Amino Acid Sequence,
pubmed-meshheading:11742126-Anti-Bacterial Agents,
pubmed-meshheading:11742126-Catalysis,
pubmed-meshheading:11742126-Cephalosporins,
pubmed-meshheading:11742126-Crystallography, X-Ray,
pubmed-meshheading:11742126-Dimerization,
pubmed-meshheading:11742126-Models, Chemical,
pubmed-meshheading:11742126-Models, Molecular,
pubmed-meshheading:11742126-Molecular Sequence Data,
pubmed-meshheading:11742126-Mutation,
pubmed-meshheading:11742126-Penicillin Amidase,
pubmed-meshheading:11742126-Protein Binding
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pubmed:year |
2002
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pubmed:articleTitle |
Structure-based prediction of modifications in glutarylamidase to allow single-step enzymatic production of 7-aminocephalosporanic acid from cephalosporin C.
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pubmed:affiliation |
Department of Biophysics, Max-Planck Institute for Medical Research, Jahnstr. 29, D-69120 Heidelberg, Germany. fritz@mpimf-heidelberg.mpg.de
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pubmed:publicationType |
Journal Article
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