The gene for indole-3-acetyl-L-aspartic acid hydrolase from Enterobacter agglomerans : molecular cloning, nucleotide sequence, and expression in Escherichia coli |
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| Authors: | J-C Chou W W Mulbry and J D Cohen |
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| Institution: | (1) Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD 20742, USA, US;(2) Soil-Microbial Systems Laboratory, Beltsville Agricultural Research Center, Agricultural Research Service, United States Department of Agriculture, Beltsville, MD 20705-2350, USA, US;(3) Horticultural Crops Quality Laboratory, Beltsville Agricultural Research Center, Agricultural Research Service, United States Department of Agriculture, Beltsville, MD 20705-2350, USA Fax: +1-301-504-5107; e-mail: jc273@umail.umd.edu, US |
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| Abstract: | A 5.5-kb DNA fragment containing the indole-3-acetyl-aspartic acid (IAA-asp) hydrolase gene (iaaspH) was isolated from Enterobacter agglomerans strain GK12 using a hybridization probe based on the N-terminal amino acid sequence of the protein. The DNA sequence of a
2.4-kb region of this fragment was determined and revealed a 1311-nucleotide ORF large enough to encode the 45-kDa IAA-asp
hydrolase. A 1.5-kb DNA fragment containing iaaspH was subcloned into the Escherichia coli expression plasmid pTTQ8 to yield plasmid pJCC2. Extracts of IPTG-induced E. coli cultures containing the pJCC2 recombinant plasmid showed IAA-asp hydrolase levels 5 to 10-fold higher than those in E. agglomerans extracts. Homology searches revealed that the IAA-asp hydrolase was similar to a variety of amidohydrolases. In addition,
IAA-asp hydrolase showed 70% sequence identity to a putative thermostable carboxypeptidase of E. coli.
Received: 12 March 1998 / Accepted: 30 March 1998 |
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| Keywords: | Hydrolase Enterobacter Amidohydrolase Carboxypeptidase Indole-3-acetylaspartic acid |
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