title: Evidence for only oxygenative cleavage of aldehydes to alk(a/e)nes and formate by cyanobacterial aldehyde decarbonylases.
abstract: Cyanobacterial aldehyde decarbonylases (ADs) catalyze the conversion of C(n) fatty aldehydes to formate (HCO(2)(-)) and the corresponding C(n-1) alk(a/e)nes. Previous studies of the Nostoc punctiforme (Np) AD produced in Escherichia coli (Ec) showed that this apparently hydrolytic reaction is actually a cryptically redox oxygenation process, in which one O-atom is incorporated from O(2) into formate and a protein-based reducing system (NADPH, ferredoxin, and ferredoxin reductase; N/F/FR) provides all four electrons needed for the complete reduction of O(2). Two subsequent publications by Marsh and co-workers [    Das, et al.     (  2011  )   Angew. Chem. Int. Ed.  50  ,   7148 - 7152  ;     Eser, et al.     (  2011  )   Biochemistry  50  ,   10743  -  10750  ] reported that their Ec-expressed Np and Prochlorococcus marinus (Pm) AD preparations transform aldehydes to the same products more rapidly by an O(2)-independent, truly hydrolytic process, which they suggested proceeded by transient substrate reduction with obligatory participation by the reducing system (they used a chemical system, NADH and phenazine methosulfate; N/PMS). To resolve this discrepancy, we re-examined our preparations of both AD orthologues by a combination of (i) activity assays in the presence and absence of O(2) and (ii) (18)O(2) and H(2)(18)O isotope-tracer experiments with direct mass-spectrometric detection of the HCO(2)(-) product. For multiple combinations of the AD orthologue (Np and Pm), reducing system (protein-based and chemical), and substrate (n-heptanal and n-octadecanal), our preparations strictly require O(2) for activity and do not support detectable hydrolytic formate production, despite having catalytic activities similar to or greater than those reported by Marsh and co-workers. Our results, especially of the (18)O-tracer experiments, suggest that the activity observed by Marsh and co-workers could have arisen from contaminating O(2) in their assays. The definitive reaffirmation of the oxygenative nature of the reaction implies that the enzyme, initially designated as aldehyde decarbonylase when the C1-derived coproduct was thought to be carbon monoxide rather than formate, should be redesignated as aldehyde-deformylating oxygenase (ADO).
authors.affiliation: Department of Biochemistry, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.
authors.forename: Ning
authors.initials: N
authors.lastname: Li
authors.forename: Wei-Chen
authors.initials: WC
authors.lastname: Chang
authors.forename: Douglas M
authors.initials: DM
authors.lastname: Warui
authors.forename: Squire J
authors.initials: SJ
authors.lastname: Booker
authors.forename: Carsten
authors.initials: C
authors.lastname: Krebs
authors.forename: J Martin
authors.initials: JM
authors.lastname: Bollinger
mesh_terms: D000446:Aldehyde-Lyases / Q000235:genetics / Q000378:metabolism*; D000447:Aldehydes / Q000737:chemistry / Q000378:metabolism*; D002853:Chromatography, Liquid; D000458:Cyanobacteria / Q000201:enzymology*; D005561:Formates / Q000378:metabolism; D006586:Hexanes / Q000737:chemistry / Q000378:metabolism; D006868:Hydrolysis; D013058:Mass Spectrometry; D010100:Oxygen / Q000737:chemistry / Q000378:metabolism*; D010103:Oxygen Isotopes / Q000737:chemistry / Q000378:metabolism
chemical_list: D000447:Aldehydes; D005561:Formates; D006586:Hexanes; D010103:Oxygen Isotopes; C046204:heptanal; D000446:Aldehyde-Lyases; C043439:aldehyde decarbonylase; D010100:Oxygen; C031639:caprylic aldehyde