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microbiology, Vol 144, 1677-1682, Copyright © 1998 by Society for General Microbiology
ARTICLES |
A Watanabe, K Yano, K Ikebukuro and I Karube
Research Center for Advanced Science and Technology, University of Tokyo, Japan.
The cyanide-degrading bacterial strain AK61 was isolated from waste water at a metal-plating plant. The isolated strain was characterized by Gram-staining, quinone analysis, fatty acid profile and the API 20NE identification system, and identified as Pseudomonas stutzeri. Whole cells were able to degrade cyanide rapidly in a 1 mM solution containing no organic substances, and produced ammonia as a product. The induction of the cyanide-degrading activity of P. stutzeri AK61 did not depend on the presence of cyanide in the culture medium during growth. The cyanide-degrading enzyme was purified approximately 49-fold from a cell extract of P. stutzeri AK61. The enzyme had a K(m) of 1.7 mM for cyanide and a specific activity of 54.6 mumol ammonia produced min-1. The activity of the enzyme was optimal at 30 degrees C and pH 7.5. The results of SDS-PAGE, gel-filtration chromatography and NH2- terminal amino acid sequence analysis of the enzyme indicated that the functional enzyme was an aggregated protein consisting of a 38 kDa polypeptide. Like cyanidase (cyanide dihydratase), it was shown that the enzyme catalysed the hydrolysis of cyanide to ammonia and formate.
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