International Journal of Agriculture, Environment & Biotechnology

Citation: IJAEB: 7 Special Issue : 409-419 July 2014

©2014 New Delhi Publishers. All rights reserved

1

Biochemistry

Comparative analysis of Polyphenol Oxidase, Catalase and

Lycopene production in Lycopersicon esculentum Mill.

Ami Lokhandwala and Madhumati Bora*

Natubhai V Patel College of Pure and Applied Sciences, Vallabh Vidyanagar–388 120, Gujarat, India.

*Corresponding author : Madhumati Bora; drmadhumatibora@gmail.com

Abstract

Antioxidant activity of tomato has been extensively studied but only in context of Lycopene. This study relates the activity of antioxidant

enzymes (Polyphenol Oxidase and Catalase) along with Lycopene in Lycopersicon esculentum Mill at different growth stages of (Seedling,

Flowering and Fruiting). Polyphenol Oxidase, Catalase (both partially purified) and Lycopene were estimated spectrophotometrically and

presence of Lycopene was further confirmed through Thin Layer and High-Performance Thin Layer Chromatography. Polyphenol Oxidase

and Catalase could be partially purified with 2.61 (22.55% yield) and 2.11 (62.3% yield) fold purification respectively via ammonium sulphate

precipitation respectively. Antioxidant enzymes showed maximum production at the seedling stage (Polyphenol Oxidases: 197.12U/ml and

Catalase: 0.037U/ml) where Lycopene production was least; while Lycopene production was maximum in the fruiting stage (259.49mg/kg

of fresh weigh) where enzyme activities were negligible. HPTLC analysis also supported the above findings. Linear Regression analysis

of Lycopene, PPO and CAT were performed in which r (correlation coefficient) value for Lycopene and PPO was -0.9279108012 and for

Lycopene and CAT was -0.7316992009; which indicated strong negative correlation between Lycopene and both the enzymes. It can be

concluded that Antioxidant enzymes play their share at young stages while Lycopene at mature stage in antioxidant network of tomato.

Highlights

Open Pollinated varieties of Lycopersicon esculentum Mill have been used for analysis.

Enzymes have been partially purified via Ammonium Sulphate precipitation followed by dialysis and their physical parameters

have been optimized.

Quantitative methods, Thin Layer chromatography and High Performance Thin Layer Chromatography have been used to study the

production of Polyphenol Oxidases, Catalase and Lycopene.

Keywords: Polyphenol Oxidase, Catalase, Lycopene, Lycopersicon esculentum Mill, Partial Purification and optimization.

Abbreviations: Polyphenol Oxidases (PPOs), Catalase (CAT)

Plantssurvivewithvariousbioticandabioticenvironmental

anti-oxidative system comprising of non-enzymatic as

stresses by altering their cellular metabolism and bringing

well as enzymatic antioxidants in plants (Noctor and

into play different defense mechanisms. Reactive species

Foyer, 1998). Antioxidants can act by rummaging reactive

are highly unstable structures; which are divided into

oxygen species (e.g. Sulphur Oxide Dismutase removing

four categories based on their central atom: Reactive

oxygen), by inhibiting their formation (e.g. by blocking

Oxygen Species (ROS), Reactive Nitrogen Species,

phagocyte activation), by binding transition metal

Reactive Chloride Species and Reactive Sulphur

ions and preventing formation of hydroxyl ions and/

Species (Halliwell and Gutteridge, 2007). Foraging or

or decomposition of lipid hydro-peroxides, by repairing

detoxification of excess ROS is attained by a competent

damage (e.g. α –tocopherol repairing peroxyl radicals and