|
type |
Journal Article |
authors |
Sheehy, R. E.; Honma, M.; Yamada, M.; Sasaki, T.; Martineau, B.; Hiatt, W. R. |
title |
Isolation, sequence, and expression in Escherichia coli of the Pseudomonas sp. strain ACP gene encoding 1-aminocyclopropane-1- carboxylate deaminase |
journal |
J Bacteriol |
Activity |
3.5.99.7 |
Family |
3.5.99.7 |
sel |
selected |
ui |
1885510 |
year |
(1991) |
volume |
173 |
number |
17 |
pages |
5260-5 |
| |
keywords |
Amino Acid Sequence |
abstract |
Pseudomonas sp. strain ACP is capable of growth on 1-aminocyclopropane- 1-carboxylate (ACC) as a nitrogen source owing to induction of the enzyme ACC deaminase and the subsequent conversion of ACC to alpha- ketobutyrate and ammonia (M. Honma, Agric. Biol. Chem. 49:567-571, 1985). The complete amino acid sequence of purified ACC deaminase was determined, and the sequence information was used to clone the ACC deaminase gene from a 6-kb EcoRI fragment of Pseudomonas sp. strain ACP DNA. DNA sequence analysis of an EcoRI-PstI subclone demonstrated an open reading frame (ORF) encoding a polypeptide with a deduced amino acid sequence identical to the protein sequence determined chemically and a predicted molecular mass of 36,674 Da. The ORF also contained an additional 72 bp of upstream sequence not predicted by the amino acid sequence. Escherichia coli minicells containing the 6-kb clone expressed a major polypeptide of the size expected for ACC deaminase which was reactive with ACC deaminase antiserum. Furthermore, a lacZ fusion with the ACC deaminase ORF resulted in the expression of active enzyme in E. coli. ACC is a key intermediate in the biosynthesis of ethylene in plants, and the use of the ACC deaminase gene to manipulate this pathway is discussed. |
last changed |
2018/05/07 13:07 |
|