Purification and Characterization of Cell Suspensions Peroxidase from Cotton (Gossypium hirsutum L.)

Abstract

Two peroxidases, cPOD-I and rPOD-II, have been isolated and purified from cotton cell suspension and their biochemical characteristics studied. rPOD-II from R405-2000, a non-embryogenic cultivar, has higher activity than cPOD-I derived from Coker 312, which developed an embryogenic structure. The cPOD-I and rPOD-II had molecular mass of 39.1 and 64 kDa respectively, as determined by SDS-PAGE. Both enzymes showed high efficiency of interaction with the guaiacol at 25 mM. The optimal pH for cPOD-I and rPOD-II activity was 5.0 and 6.0, respectively. The enzyme had an optimum temperature of 25 degrees C and was relatively stable at 20-30 degrees C. The isoenzymes were highly inhibited by ascorbic acid, dithiothreitol, sodium metabisulfite, and beta-mercaptoethanol. Their activities were highly enhanced by Al(3+), Fe(3+), Ca(2+), and Ni(2+), but they were moderately inhibited by Mn(2+) and K(+). The enzyme lost 50% to 62% of its activity in the presence of Zn(2+) and Hg(2+).

Partial Purification and Characterization of Peroxidase from Pearl Millet (Pennisetum Glaucum [L.] R. Br.) Grains

“…However, the determined molecular mass is lower than that reported for the enzyme from Raphanus sativus seeds (44 kDa; Kim and Lee ), cotton seeds (64 kDa; Kouakou et al . ) and radish (66 kDa; Şişecioğlu et al . ).…”

“…), cotton (Kouakou et al . ), black gram (Ajila and Rao ), wheat (Manu and Rao ), pearl millet seedlings (Devaiah and Shetty ), radish (Şişecioğlu et al . ), Oryza sativa (Verma et al .…”

A Defense Associated Peroxidase from Lemon Having Dye Decolorizing Ability and Offering Resistance to Heat, Heavy Metals and Organic Solvents

“…On the other hand, Hg 2+ exhibited slight inhibitory effect on the purified peroxidase at all the concentrations studied. [33], the Leucaena peroxidase has also reported to be activated by Na + , Ca 2+ and Mn 2+ [40] and that of cotton peroxidases are also reported to be activated by Ni +2 [25]. On the other hand, other peroxidases such as the lettuce peroxidase was apparently inhibited by metal ions namely, Zn 2+ , Ca 2+ and Mn 2+ [15], the Cs 2+ and Ni 2+ have reported to show inhibitory effect on Ipomoea carnea peroxides [41].…”

“…[25][26][27][28][29]. Native structure of plant peroxidases are reported to vary from being monomeric to multimeric (either homo-or hetero-Page 2 of 10 meric).…”

“…For example, the African oil palm tree peroxidase which exhibited stability at 70°C for 1 hr at neutral pH [37], while, the Ficus sycomorus peroxidase retained 80% activity at 70°C after 15 min incubation [33] and the peroxidase isolated from vigna species retained its 65% activity for 1 hr at 70°C [35]. The horseradish peroxidase (HRP), the industrially most exploited peroxidase, exhibited loss of activity at 70°C within 5 min [25]. Thus, the availability of novel peroxidases having tolerance towards heat, organic solvents, salts, heavy metals etc, is scarce.…”