title: Phenoloxidase activity of intact and chemically modified functional unit RvH1: a from molluscan Rapana venosa hemocyanin.
abstract: o-Diphenol oxidase activities (o-diPO) of chemically modified functional unit RvH1-a of molluscan hemocyanin Rapana venosa were studied using L-Dopa and dopamine as substrates. With L-Dopa as substrate the native FU RvH1-a did not show any o-diPO activity. Therefore the native FU RvH1-a was converted to enzymatic active form, after treatment with SDS, trypsin, urea and different values of pH when its o-diPO activity was studied. The highest artificial induction of o-diPO activity was observed after incubation of FU with 3.0mM SDS, and RvH1-a shows both, dopamine (K(M)=6.53mM, k(cat)/K(M)=1.29) and L-Dopa (K(M)=2.0mM, k(cat)/K(M)=2.1) activity due to a more open active site of the enzyme and better access of the substrates. It was determined that the K(M) value of SDS-activated RvH1-a against dopamine is higher compared to those of hemocyanins from Helix vulgaris, Helix pomatia and native tyrosinase from Ipomoea batatas but much lower than that from Illex argentinus (ST94) tyrosinase and arthropodan hemocyanin from Carcinus aestuarii. The Km value of SDS-activated RvH1-a against L-Dopa is higher than those of hemocyanins from H. vulgaris and Cancer magister, but lower than that of the tyrosinase from Streptomyces albus.
authors.affiliation: Institute of Organic Chemistry with Center of Phytochemistry, Bulgarian Academy of Sciences, Sofia. adolashki@yahoo.com
authors.forename: Aleksandar
authors.initials: A
authors.lastname: Dolashki
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Voelter
authors.forename: Pavlina
authors.initials: P
authors.lastname: Dolashka
mesh_terms: D000818:Animals; D004298:Dopamine / Q000378:metabolism; D049851:Gastropoda / Q000201:enzymology*; D006372:Helix, Snails / Q000201:enzymology; D006433:Hemocyanins / Q000737:chemistry*; D019476:Insect Proteins / Q000737:chemistry / Q000378:metabolism; D007980:Levodopa / Q000378:metabolism; D014442:Monophenol Monooxygenase / Q000737:chemistry* / Q000378:metabolism*; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism*
chemical_list: D019476:Insect Proteins; D021122:Protein Subunits; D007980:Levodopa; D006433:Hemocyanins; D014442:Monophenol Monooxygenase; D004298:Dopamine