title: Purification and characterization of Mn-peroxidase from Musa paradisiaca (banana) stem juice.
abstract: Mn-peroxidase (MnP), a biotechnologically important enzyme was purified for the first time from a plant source Musa paradisiaca (banana) stem, which is an agro-waste easily available after harvest of banana fruits. MnP was earlier purified only from the fungal sources. The enzyme was purified from stem juice by ultrafiltration and anion-exchange column chromatography on diethylamino ethylcellulose with 8-fold purification and purification yield of 65%. The enzyme gave a single protein band in SDS-PAGE corresponding to molecular mass 43 kDa. The Native-PAGE of the enzyme also gave a single protein band, confirming the purity of the enzyme. The UV/VIS spectrum of the purified enzyme differed from the other heme peroxidases, as the Soret band was shifted towards lower wavelength and the enzyme had an intense absorption band around 250 nm. The K(m) values using MnSO4 and H2O2 as the substrates of the purified enzyme were 21.0 and 9.5 microM, respectively. The calculated k(cat) value of the purified enzyme using Mn(II) as the substrate in 50 mM lactate buffer (pH 4.5) at 25 degrees C was 6.7s(-1), giving a k(cat)/K(m) value of 0.32 microM(-1)s(-1). The k(cat) value for the MnP-catalyzed reaction was found to be dependent of the Mn(III) chelator molecules malonate, lactate and oxalate, indicating that the enzyme oxidized chelated Mn(II) to Mn(III). The pH and temperature optima of the enzyme were 4.5 and 25 degrees C, respectively. The enzyme in combination with H2O2 liberated bromine and iodine in presence of KBr and KI respectively. All these enzymatic characteristics were similar to those of fungal MnP. The enzyme has the potential as a green brominating and iodinating agent in combination with KBr/KI and H2O2.
authors.affiliation: Department of Chemistry, U P P G College, Varanasi 221002 (U. P.), India.
authors.forename: Pratibha
authors.initials: P
authors.lastname: Yadav
authors.forename: V K
authors.initials: VK
authors.lastname: Singh
authors.forename: Meera
authors.initials: M
authors.lastname: Yadav
authors.forename: Sunil Kumar
authors.initials: SK
authors.lastname: Singh
authors.forename: Sudha
authors.initials: S
authors.lastname: Yadava
authors.forename: K D S
authors.initials: KD
authors.lastname: Yadav
mesh_terms: D002384:Catalysis; D002848:Chromatography, DEAE-Cellulose; D004795:Enzyme Stability; D054879:Halogenation; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D028521:Musa / Q000201:enzymology*; D010084:Oxidation-Reduction; D010544:Peroxidases / Q000737:chemistry* / Q000302:isolation & purification* / Q000493:pharmacokinetics; D010936:Plant Extracts / Q000302:isolation & purification; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000493:pharmacokinetics; D018547:Plant Stems / Q000201:enzymology; D013056:Spectrophotometry, Ultraviolet; D013379:Substrate Specificity; D013696:Temperature; D014462:Ultrafiltration
chemical_list: D010936:Plant Extracts; D010940:Plant Proteins; D010544:Peroxidases; C051129:manganese peroxidase