Biochemical and molecular characterization of a ring fission dioxygenase with the ability to oxidize (substituted) salicylate(s) from Pseudaminobacter salicylatoxidans.

The Journal of Biological Chemistry
J P HintnerAndreas Stolz

Abstract

The gene coding for a dioxygenase with the ability to cleave salicylate by a direct ring fission mechanism to 2-oxohepta-3,5-dienedioic acid was cloned from Pseudaminobacter salicylatoxidans strain BN12. The deduced amino acid sequence encoded a protein with a molecular mass of 41,176 Da, which showed 28 and 31% sequence identity, respectively, to a gentisate 1,2-dioxygenase from Pseudomonas alcaligenes NCIMB 9867 and a 1-hydroxy-2-naphthoate 1,2-dioxygenase from Nocardioides sp. KP7. The highest degree of sequence identity (58%) was found to a presumed gentisate 1,2-dioxygenase from Corynebacterium glutamicum. The enzyme from P. salicylatoxidans BN12 was heterologously expressed in Escherichia coli and purified as a His-tagged enzyme variant. The purified enzyme oxidized in addition to salicylate, gentisate, 5-aminosalicylate, and 1-hydroxy-2-naphthoate also 3-amino- and 3- and 4-hydroxysalicylate, 5-fluorosalicylate, 3-, 4-, and 5-chlorosalicylate, 3-, 4-, and 5-bromosalicylate, 3-, 4-, and 5-methylsalicylate, and 3,5-dichlorosalicylate. The reactions were analyzed by high pressure liquid chromatography/mass spectrometry, and the reaction products were tentatively identified. For comparison, the putative gentisate 1,2-dioxyge...Continue Reading

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Citations

Aug 30, 2012·Dalton Transactions : an International Journal of Inorganic Chemistry·Amanda E BaumAdam T Fiedler
Jun 7, 2006·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·I MateraF Briganti
Jan 19, 2010·Applied and Environmental Microbiology·Yi Qu, Jim C Spain
Jan 31, 2012·Applied and Environmental Microbiology·Susanne Fetzner
Jan 11, 2007·Bioscience, Biotechnology, and Biochemistry·Shin-ichi HiranoShigenori Kanaya
Jun 14, 2012·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Stefano Da VelaFabrizio Briganti
Nov 20, 2009·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Timothy E MachonkinSara S Rocks
Jul 29, 2015·Applied Microbiology and Biotechnology·Martina Kiel, Karl-Heinrich Engesser
Aug 4, 2019·Environmental Science and Pollution Research International·Estefania BraccoRoberto Candal
Feb 4, 2019·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Aleksey AleshintsevRupal Gupta

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