Purification, properties, and distribution of ascorbate peroxidase in legume root nodules

Journal Article · Thu Jan 01 00:00:00 EST 1987 · Plant Physiology; (USA)

DOI:https://doi.org/10.1104/pp.83.4.789· OSTI ID:5081109

Dalton, D A; Hanus, F J; Russell, S A; Evans, H J ^[1]

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1. Oregon State Univ., Corvallis (USA)

All aerobic biological system, including N{sub 2}-fixing root nodules, are subject to O{sub 2} toxicity that results from the formation of reactive intermediates such as H{sub 2}O{sub 2} and free radicals of O{sub 2}. H{sub 2}O{sub 2} may be removed from root nodules in a series of enzymic reactions involving ascorbate peroxidase, dehydroascorbate reductase, and glutathione reductase. The authors confirm here the presence of these enzymes in root nodules from nine species of legumes and from Alnus rubra. Ascorbate peroxidase from soybean nodules was purified to near homogeneity. This enzyme was found to be a hemeprotein with a molecular weight of 30,000 as determined by sodium dodecyl sulfate gel electrophoresis. KCN, NaN{sub 3}, CO, and C{sub 2}H{sub 2} were potent inhibitors of activity. Nonphysiological reductants such as guaiacol, o-dianisidine, and pyrogallol functioned as substrates for the enzyme. No activity was detected with NAD(P)H, reduced glutathione, or urate. Ascorbate peroxidation did not follow Michaelis-Menten kinetics. The substrate concentration which resulted in a reaction rate of 1/2 V{sub max} was 70 micromolar for ascorbate and 3 micromolar for H{sub 2}O{sub 2}. The high affinity of ascorbate peroxidase for H{sub 2}O{sub 2} indicates that this enzyme, rather than catalase, is responsible for most H{sub 2}O{sub 2} removal outside of peroxisomes in root nodules.

• OSTI ID: 5081109
• Journal Information: Plant Physiology; (USA), Vol. 83:4; ISSN 0032-0889
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
PEROXIDASES
ENZYME ACTIVITY
ROOTS
ALFALFA
BIOCHEMICAL REACTION KINETICS
CHEMICAL PROPERTIES
CLOVER
ENZYME INHIBITORS
NITROGEN FIXATION
PEANUTS
PISUM
SUBSTRATES
VICIA
ENZYMES
KINETICS
LEGUMINOSAE
MAGNOLIOPHYTA
MAGNOLIOPSIDA
OXIDOREDUCTASES
PLANTS
REACTION KINETICS
SEEDS
553004* - Agriculture & Food Technology- Food Protection & Preservation- (1987-)