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.
Original language | English (US) |
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Pages (from-to) | 172-178 |
Number of pages | 7 |
Journal | Molecular and General Genetics |
Volume | 259 |
Issue number | 2 |
DOIs | |
State | Published - 1998 |
Bibliographical note
Funding Information:Acknowledgements This work was supported by grant DE-A102-94ER20153 from the US Department of Energy.
Keywords
- Amidohydrolase carboxypeptidase
- Enterobacter
- Hydrolase
- Indole-3-acetylaspartic acid