title: Purification and characterization of NAD+ -dependent salicylaldehyde dehydrogenase from carbaryl-degrading Pseudomonas sp. strain C6.
abstract: NAD+-dependent salicylaldehyde dehydrogenase (SALDH) which catalyzes the oxidation of salicylaldehyde to salicylate was purified form carbaryl-degrading Pseudomonas sp. strain C6. The enzyme was found to be a functional homotrimer (150 kDa) with subunit molecular mass of 50 kDa and contained calcium (1.8 mol/mol of enzyme). These properties were found to be unique. External addition of metal ions showed no effect on the activity and addition of chelators showed moderate inhibition of the activity. Potassium ions were found to enhance the activity significantly. SALDH showed higher affinity for salicylaldehyde (Km = 4.5 μM) and accepts mono- as well as di-aromatic aldehydes; however it showed poor activity on aliphatic aldehydes. Chloro-/nitro-substituted benzaldehydes were potent substrate inhibitors as compared to benzaldehyde and 3-hydroxybenzaldehyde, while 2-naphthaldehyde and salicylaldehyde were moderate. The kinetic data revealed that SALDH, though having broad specificity, is more efficient for the oxidation of salicylaldehyde as compared to other aromatic aldehyde dehydrogenases which gives an advantage for Pseudomonas sp. strain C6 to bioremediate carbaryl and other aromatic aldehydes efficiently.
authors.forename: Randhir
authors.initials: R
authors.lastname: Singh
authors.forename: Vikas D
authors.initials: VD
authors.lastname: Trivedi
authors.forename: Prashant S
authors.initials: PS
authors.lastname: Phale
mesh_terms: D000445:Aldehyde Oxidoreductases / Q000302:isolation & purification* / Q000378:metabolism*; D001673:Biodegradation, Environmental / Q000187:drug effects; D012721:Carbaryl / Q000378:metabolism*; D007700:Kinetics; D008970:Molecular Weight; D009243:NAD / Q000378:metabolism*; D011188:Potassium / Q000494:pharmacology; D011549:Pseudomonas / Q000187:drug effects / Q000201:enzymology*; D012964:Sodium / Q000494:pharmacology; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D013379:Substrate Specificity / Q000187:drug effects; D013997:Time Factors
chemical_list: D009243:NAD; D012964:Sodium; D000445:Aldehyde Oxidoreductases; C024246:salicylaldehyde dehydrogenase; D012721:Carbaryl; D011188:Potassium