Characterization of the Pseudomonas pseudoalcaligenes CECT5344 Cyanase, an enzyme that is not essential for cyanide assimilation.

Applied and Environmental Microbiology
V M Luque-AlmagroR Blasco

Abstract

Cyanase catalyzes the decomposition of cyanate into CO(2) and ammonium, with carbamate as an unstable intermediate. The cyanase of Pseudomonas pseudoalcaligenes CECT5344 was negatively regulated by ammonium and positively regulated by cyanate, cyanide, and some cyanometallic complexes. Cyanase activity was not detected in cell extracts from cells grown with ammonium, even in the presence of cyanate. Nevertheless, a low level of cyanase activity was detected in nitrogen-starved cells. The cyn gene cluster of P. pseudoalcaligenes CECT5344 was cloned and analyzed. The cynA, cynB, and cynD genes encode an ABC-type transporter, the cynS gene codes for the cyanase, and the cynF gene encodes a novel sigma(54)-dependent transcriptional regulator which is not present in other bacterial cyn gene clusters. The CynS protein was expressed in Escherichia coli and purified by following a simple and rapid protocol. The P. pseudoalcaligenes cyanase showed an optimal pH of 8.5 degrees C and a temperature of 65 degrees C. An insertion mutation was generated in the cynS gene. The resulting mutant was unable to use cyanate as the sole nitrogen source but showed the same resistance to cyanate as the wild-type strain. These results, in conjunction wi...Continue Reading

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Citations

Jul 19, 2012·World Journal of Microbiology & Biotechnology·Luigia SabatiniFrancesca Bruscolini
Jan 9, 2016·Current Opinion in Biotechnology·Víctor M Luque-AlmagroMaría Dolores Roldán
Mar 30, 2010·Journal of Hazardous Materials·M J HuertasI García-García
Sep 25, 2012·Environmental Microbiology·Víctor M Luque-AlmagroRafael Blasco
Sep 20, 2015·Parasites & Vectors·Reza Zolfaghari EmamehSeppo Parkkila
Jan 27, 2011·Biochemical Society Transactions·Victor M Luque-AlmagroM Dolores Roldán
Jul 30, 2015·Nature·Marton PalatinszkyMichael Wagner
Jun 15, 2017·Journal of Agricultural and Food Chemistry·Caleb R SchlachterMaksymilian Chruszcz
May 29, 2018·Applied and Environmental Microbiology·Reza Zolfaghari EmamehSeppo Parkkila
Mar 28, 2017·Environmental Science and Pollution Research International·Rashad Kebeish, Omar Al-Zoubi
Dec 14, 2018·World Journal of Microbiology & Biotechnology·Tomas Linder
Apr 24, 2019·World Journal of Microbiology & Biotechnology·Monica SharmaSubhankar Chatterjee
Feb 23, 2020·Applied Microbiology and Biotechnology·Lynn Sophie SchwardmannSkander Elleuche
May 18, 2019·Environmental Science and Pollution Research International·Xiao-Zhang YuQing Zhang
Nov 12, 2018·Applied Microbiology and Biotechnology·Hanna KochSebastian Lücker
May 9, 2021·Molecular Biology and Evolution·Fiona J WhelanJames O McInerney

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