3-Carboxy-cis,cis-muconate lactonizing enzyme from Pseudomonas putida is homologous to the class II fumarase family: a new reaction in the evolution of a mechanistic motif
Autor: | Stephen C. Ransom, John W. Kozarich, Suzanne E. Williams, Patricia C. Babbitt, Elisa M. Woolridge, James A. Landro |
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Rok vydání: | 1992 |
Předmět: |
DNA
Bacterial Base Sequence Sequence Homology Amino Acid Pseudomonas putida Molecular Sequence Data Nucleic acid sequence Bacillus subtilis Biology Molecular cloning biology.organism_classification Biological Evolution Biochemistry Homology (biology) Fumarate Hydratase Kinetics Fumarase Escherichia coli Electrophoresis Polyacrylamide Gel Amino Acid Sequence Cloning Molecular Adenylosuccinate lyase Peptide sequence Plasmids |
Zdroj: | Biochemistry. 31:9768-9776 |
ISSN: | 1520-4995 0006-2960 |
Popis: | The gene (pcaB) for 3-carboxymuconate lactonizing enzyme (CMLE; 3-carboxymuconate cycloisomerase; EC 5.5.1.2) from Pseudomonas putida has been cloned into pMG27NS, a temperature-sensitive expression vector, and expressed in Escherichia coli N4830. The specific activity and kinetic parameters of the recombinant CMLE were comparable to those previously reported. A comparison of the deduced amino acid sequence of CMLE with sequences available in the PIR and Genbank databases revealed that CMLE has highly significant sequence homology to the class II fumarase family, particularly to adenylosuccinate lyase from Bacillus subtilis. CMLE has no significant homology to muconate lactonizing enzyme (MLE) from P. putida, its sister enzyme in the beta-ketoadipate pathway. These findings fully corroborate a prediction made by us on the basis of mechanistic and stereochemical analyses of CMLE and MLE [Chari, R. V. J., Whitman, C. P., Kozarich, J. W., Ngai, K.-L., & Ornston, L. N. (1987) J. Am. Chem. Soc. 109, 5514-5519] and suggest that CMLE and MLE were recruited into this specialized pathway from two different enzyme families. |
Databáze: | OpenAIRE |
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