Cloning, sequencing, and expression of the genes encoding the adenosylcobalamin-dependent ethanolamine ammonia-lyase of Salmonella typhimurium

Ethanolamine ammonia-lyase is a bacterial enzyme that catalyzes the adenosylcobalamin-dependent conversion of certain vicinal amino alcohols to oxo compounds and ammonia. Studies of ethanolamine ammonia-lyase from Clostridium sp. and Escherichia coli have suggested that the enzyme is a heterodimer c...

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Bibliographic Details
Published in:The Journal of biological chemistry 1990-07, Vol.265 (21), p.12462-12466
Main Authors: FAUST, L. R. P, CONNOR, J. A, ROOF, D. M, HOCH, J. A, BABIOR, B. M
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Ammonia-Lyases - genetics
Base Sequence
Biological and medical sciences
Cloning, Molecular
DNA, Bacterial - genetics
Ethanolamine Ammonia-Lyase - genetics
Fundamental and applied biological sciences. Psychology
Gene Expression Regulation, Bacterial
genes
Genes, Bacterial
Genes. Genome
Genetic Complementation Test
Molecular and cellular biology
Molecular genetics
Molecular Sequence Data
RNA, Messenger - genetics
Salmonella typhimurium
Salmonella typhimurium - enzymology
Salmonella typhimurium - genetics
Transcription, Genetic
Vitamin B 12 - metabolism
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Summary:Ethanolamine ammonia-lyase is a bacterial enzyme that catalyzes the adenosylcobalamin-dependent conversion of certain vicinal amino alcohols to oxo compounds and ammonia. Studies of ethanolamine ammonia-lyase from Clostridium sp. and Escherichia coli have suggested that the enzyme is a heterodimer composed of subunits of Mr approximately 55,000 and 35,000. Using a partial Sau3A Salmonella typhimurium library ligated into pBR328 and selecting by complementation of a mutant lacking ethanolamine ammonia-lyase activity, we have cloned the genes for the 2 subunits of the S. typhimurium enzyme. The genes were localized to a 6.5-kilobase fragment of S. typhimurium DNA, from which they could be expressed in E. coli under noninducing conditions. Sequencing of a 2526-base pair portion of this 6.5-kilobase DNA fragment revealed two open reading frames separated by 21 base pairs. The open reading frames encoded proteins of 452 and 286 residues whose derived N-terminal sequences were identical to the N-terminal sequences of the 2 subunits of the E. coli ethanolamine ammonia-lyase, except that residue 16 of the large subunit was asparagine in the E. coli sequence and aspartic acid in the S. typhimurium sequence.
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(19)38368-1