Purification, characterization and primary structure determination of aldoxime dehydratase

Purification, characterization and primary structure determination of aldoxime dehydratase  

With these results, we named the new enzyme as phenylacetaldoxime dehydratase (EC 4.99.1.7). It was also suggested that the enzyme utilizes FMN as an electron acceptor, because the value was increased about five times under anaerobic condition and the sulfite ion could replace FMN, although the enzyme requires oxidized form of FMN. It was revealed that the enzyme is a quite unique enzyme whose apparent function is to catalyze a dehydration reaction. The reaction mechanism is of much interest. 

The gene for the dehydratase was expressed in E. coli under lac promoter, and an expression plasmid pOxD 90F was constructed. The transformant was cultivated under optimal condition at 30° C when much of the enzyme was expressed in a soluble form with more than thousand and several hundred times than the wild-type strain per culture (up to more than 50% of the total soluble protein of the 

We also found that the occurrence of aldoxime dehydratase is as wide as that for nitrile-degrading enzymes such as nitrile hydratase, amidase and/or nitri-lase. All of the nitrile degraders hitherto isolated contained aldoxime dehydratase activities. The author would like to propose that the pathway in which aldoximes are successively degraded via nitrile could be named as the aldoxime-nitrile pathway (Fig. 1). 

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