Purification of Cyanide-Degrading Nitrilase from Pseudomonas Fluorescens NCIMB 11764. Page: 36
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CHAPTER IV
DISCUSSION
This project was initiated for the purpose of identifying the enzyme from P.
fluorescens NCIMB 11764 shown previously to be capable of degrading cyanide
independently but also capable of joining with several other enzymes to produce a
novel enzyme complex with high cyanide scavenging ability (Fernandez and Kunz,
2005). The latter complex known as CNO has important physiological implications
because of its ability to serve as an efficient cyanide detoxification system (micromolar
range) and also because it is known to be required for utilization of cyanide as the sole
nitrogen source. Previous studies suggested that the enzyme component of CNO
capable of attacking cyanide on its own (but only at much higher concentrations then
when combined with other CNO partners) was a likely cyanide dihydratase (CynD).
These enzymes represent a small subset of enzymes grouped in branch 1 of the large
superfamily of enzymes classified as nitrilases (E.C. 3.5.53) that act on cyanide
specifically, catalyzing its direct hydrolysis to ammonia and formic acid. Besides
CynD enzymes, branch 1 includes many enzymes that catalyze a similar conversion of
higher nitriles referred to in general, as nitrilases (Nit) (Pace and Brenner, 2001).
Extensive efforts in our laboratory to verify that the enzyme in question was indeed
related to other CynD enzymes from bacteria were unsuccessful. Instead, while the
enzyme was shown to fall into the same branch of the superfamily as CynD's (branch
1) it was not sufficiently homologous to warrant its being considered an ortholog
thereof. Nonetheless, for reasons already discussed there was reason to think that36
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Chou, Chia-Ni. Purification of Cyanide-Degrading Nitrilase from Pseudomonas Fluorescens NCIMB 11764., thesis, December 2010; Denton, Texas. (https://digital.library.unt.edu/ark:/67531/metadc33224/m1/46/: accessed May 6, 2024), University of North Texas Libraries, UNT Digital Library, https://digital.library.unt.edu; .