Skip to content
Research Article Open access CC BY 4.0

Evaluation of Inactivation Kinetics and Thermodynamic Properties of Peroxidase from Cucumis sativus for Biotechnological and Industrial Applications

Iweha Christian Chukwunyere, Ugwoke Faith Ifeanyi, Ogara Lydia Amaechi, Nweze Justus Amuche, Ogbonna Kenneth Ejike, Chinedum Patricia Eluka, Akor Joseph, Odu Maureen Nwamaka, Joshua Parker Elijah

Journal of Advances in Biology & Biotechnology · pp. 46–62 · Published 28 Jul 2021

10.9734/jabb/2021/v24i630221

Abstract

The unpurified enzyme gave specific activity of 51.36 μ/mg and thereafter it was subjected to two phases of purification process of salt precipitation and gel filtration. The precipitating agent was ammonium sulphate while Sephadex-G100 served as purification matrix. The purification fold achieved after precipitation and filtration was 3.3 and with corresponding specific activities of 34.22 μ/mg and 116.31 μ/mg. The substrate used for the assay was o-dianisidine. Within 40-80°C of temperature, the kinetics of the peroxidase inactivation was evaluated. The results from assays showed that cucumber peroxidase conformed to the hypothesis of  Michealis-Menten Theory. From the Lineweaver-Burk plot, Michaelis Constant (Km) and maximum velocity (Vmax) were evaluated and  obtained 5.02mg/ml and 11.57μmol/min respectively. The heat induced inactivation gave biphasic curves, where initial rise in temperature was rapidly succeeded with much slower decrease. A first-order kinetic behaviour was observed for cucumber peroxidase heat inactivation. The k values of between 3.49×10-2 to 8.38×10-2 min-1 was obtained while the Z value was found to be 22.3°C. Decrease in k values with rise in temperature suggests that cucumber peroxidase was rapidly inactivated at elevated temperature. The slope of Arrhenius plot gave the activation energy of 127.99KJMol-1K-1. Also evaluated were thermodynamic constants (ΔH, G Δ, ΔS) for inactivation of peroxidase at variable temperatures. Cucumber peroxidase activity was observed to be pH sensitive and stable within pH range of 5.6-8. Further decrease or increase from this range resulted to decrease in peroxidase stability.

Concentration enzyme peroxidase cucumber homogenization industrial bioremediation

References (21)

  1. 1 PROTEIN MEASUREMENT WITH THE FOLIN PHENOL REAGENT [DOI]
  2. 2 Physiological and Molecular Plant Pathology
  3. 3 Characterization of Polyphenol Oxidase and Peroxidase and Influence on Browning of Cold Stored Strawberry Fruit [DOI]
  4. 4 Thermostability of crude endoglucanase from Aspergillus fumigatus grown under solid state fermentation (SSF) and submerged fermentation (SmF) [DOI]
  5. 5 Recent Advances in Chemistry of Enzymatic Browning [DOI]
  6. 6 Effect of Heat Treatment on Bovine Lactoperoxidase Activity in Skim Milk: Kinetic and Thermodynamic Analysis [DOI]
  7. 7 Comparative Studies on Nutritional Composition of Four Melon Seeds Varieties [DOI]
  8. 8 What is the role of thermodynamics on protein stability? [DOI]
  9. 9 Properties of a Cationic Peroxidase from Citrus jambhiri cv. Adalia [DOI]
  10. 10 Physical and enzymatic properties of lignin peroxidase isoenzymes from Phanerochaete chrysosporium [DOI]
  11. 11 Stability and identification of active-site residues of carboxymethylcellulases fromAspergillus niger andCellulomonas biazotea [DOI]
  12. 12 Purification and thermodynamic characterization of glucose oxidase from a newly isolated strain ofAspergillus niger [DOI]
  13. 13 Purification and partial characterization of peroxidase from lettuce stems [DOI]
  14. 14 Purification and Characterization of Lignin Peroxidase Isozymes from Humicola grisea (Traaen) and Its Application in Bioremediation of Textile Dyes [DOI]
  15. 15 Thermal Stabilities of Peroxidases from Fresh Pinto Beans [DOI]
  16. 16 Rapid and direct spectrophotometric method for kinetics studies and routine assay of peroxidase based on aniline diazo substrates [DOI]
  17. 17 The Contribution of an Ionic Peroxidase Isozyme to Enzyme-Mediated Browning in Dioscorea esculenta L. Tubers [DOI]
  18. 18 Journal of Agricultural and Food Chemistry [DOI]
  19. 19 International Food Research Journal [DOI]
  20. 20 Journal of Food Science and Technology [DOI]
  21. 21 International Journal of Agriculture and Biology

Cited by 1

1 citation reported by external sources — individual citing-article records aren't available to list yet.

Related research

Article metrics

Real usage data collected on this platform.

28

Page views

0

PDF downloads

2

Outbound clicks

1

Citations

Views over time

Views by country

Approximate, from request IP at view time — not citizenship or institution. Countries with fewer than 5 views are grouped as "Other".

Traffic sources

Referring site, by host.

Views and downloads exclude known bots/crawlers. Citations combines this platform's own DOI-resolved index with each external source's own reported total — see Cited by above for individually listed citing works. Last refreshed 0 seconds ago.