title: Single-step purification of peroxidase by 4-aminobenzohydrazide from Turkish blackradish and Turnip roots.
abstract: Peroxidases (PODs) were purified from the Turkish blackradish (Raphanus sativus L.) (TBR) and Turnip (Brassica rapa L.) using a simple and effective single-step method. An affinity resin was synthesised by coupling the 4-aminobenzohydrazide ligand and the l-tyrosine spacer-arm to CNBr-activated-Sepharose-4B. The purification factors for the TBR-POD and the Turnip-POD were 40.3-fold (with a yield of 10.6%) and 269.3-fold (with a yield of 9%), respectively. The molecular masses of the TBR-POD and Turnip-POD were approximately 67.3 and 65.8kDa, respectively. For guaiacol, the Km and Vmax values were calculated as 24.88mM and 3.23EU/mL, respectively for TBR-POD and as 4.09mM and 0.797EU/mL for the Turnip-POD. For H2O2, the Km and Vmax values were calculated as 3.247mM and 0.799EU/mL, respectively for TBR-POD, and as 12.49mM and 4.055EU/mL, respectively for the Turnip-POD. Furthermore, 4-aminobenzohydrazide was determined to be a non-competitive inhibitor of TBR-POD and Turnip-POD.
authors.affiliation: Faculty of Sciences, Department of Chemistry, Ataturk University, 25240 Erzurum, Turkey.
authors.forename: Ramazan
authors.initials: R
authors.lastname: Kalin
authors.affiliation: Ispir Hamza Polat Vocational Training School, Ataturk University, 25900 Erzurum, Turkey.
authors.forename: Ali
authors.initials: A
authors.lastname: Atasever
authors.affiliation: Faculty of Sciences, Department of Chemistry, Ataturk University, 25240 Erzurum, Turkey. Electronic address: hozdemir@atauni.edu.tr.
authors.forename: Hasan
authors.initials: H
authors.lastname: Özdemir
mesh_terms: D029688:Brassica napus / Q000737:chemistry / Q000201:enzymology*; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D006834:Hydrazines / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D009195:Peroxidase / Q000737:chemistry / Q000302:isolation & purification*; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification*; D018517:Plant Roots / Q000737:chemistry / Q000201:enzymology*; D031224:Raphanus / Q000737:chemistry / Q000201:enzymology*; D012685:Sepharose / Q000737:chemistry; D014421:Turkey
chemical_list: D006834:Hydrazines; D010940:Plant Proteins; D012685:Sepharose; D009195:Peroxidase
keywords: Affinity chromatography; Inhibition; Kinetics; Peroxidase