Characterisation of phenol oxidase and peroxidase from maize silk.
Silk of some maize genotypes contains a high level of phenolics that undergo enzymatic oxidation to form quinones, which condense among themselves or with proteins to form brown pigments. Two phenolic oxidizing enzymes, peroxidase (POD; EC 1.11.1.7) and polyphenol oxidase (PPO; EC 1.10.3.1), from maize (Zea mays L.) silk were characterised with respect to their preferred substrate, different isoforms and specific effectors. One browning silk sample with high, and two non-browning samples with low phenolic content were investigated. Although POD oxidizes a wide range of phenolic substrates in vitro, its activity rate was independent of silk phenolic content. PPO activity, detected with o-diphenolic substrates, was abundant only in browning silk, and low or absent in non-browning silk. Pollination increased POD but not PPO activity. Isoelectric-focusing (IEF) and specific staining for POD and PPO showed a high degree of polymorphism that varied with silk origin. The IEF pattern of POD revealed a number of anionic and several cationic isoenzymes, with the most pronounced having neutral pI 7 and a basic isoform with pI 10. Detected isoforms of PPO were anionic, except for one neutral form found only in browning silk, and occupied positions different from those of POD. Different inhibitory effects of NaN(3), EDTA, KCN, and L-cysteine, as well as different impacts of a variety of cations on the oxidation of chlorogenic acid, mediated by PPO or POD, were detected. The findings are discussed in terms of a possible roles of these enzymes in defence and pollination.
Sukalović
V Hadzi-Tasković
VH
Institute for Multidisciplinary Research, Belgrade, Serbia. hvesna@mrizp.co.rs
Veljović-Jovanović
S
S
Maksimović
J Dragisić
JD
Maksimović
V
V
Pajić
Z
Z
D004156:Catechol Oxidase / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D005419:Flavonoids / Q000032:analysis; D007527:Isoenzymes / Q000378:metabolism; D010084:Oxidation-Reduction; D009195:Peroxidase / Q000378:metabolism*; D010636:Phenols / Q000032:analysis*; D054817:Pollination; D013379:Substrate Specificity; D003313:Zea mays / Q000737:chemistry / Q000201:enzymology*
D005419:Flavonoids; D007527:Isoenzymes; D010636:Phenols; D004156:Catechol Oxidase; D009195:Peroxidase