Isolation, hyperexpression, and sequencing of the aceA gene encoding isocitrate lyase in Escherichia coli
- Washington State Univ., Pullman (USA)
A structural gene for isocitrate lyase was isolated from a cosmid containing an ace locus of the Escherichia coli chromosome. Cloning and expression under control of the tac promoter in a multicopy plasmid showed that a 1.7-kilobase-pair DNA segment was sufficient for complementation of an aceA deletion mutation and overproduction of isocitrate lyase. DNA sequence analysis of the cloned gene and N-terminal protein sequencing of the cloned and wild-type enzymes revealed an entire aceA gene which encodes a 429-amino-acid residue polypeptide whose C-terminus is histidine. The deduced amino acid sequence for the 47.2-kilodalton subunit of E. coli isocitrate lyase could be aligned with that for the 64.8-kilodalton subunit of the castor bean enzyme with 39% identity except for limited N- and C-terminal regions and a 103-residue stretch that was unique for the plant enzyme and started approximately in the middle of that peptide.
- DOE Contract Number:
- FG06-88ER13923
- OSTI ID:
- 6969057
- Journal Information:
- Journal of Bacteriology; (USA), Journal Name: Journal of Bacteriology; (USA) Vol. 170:10; ISSN JOBAA; ISSN 0021-9193
- Country of Publication:
- United States
- Language:
- English
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