Identification of coding sequence and its use for functional and structural characterization of catalase from Phyllanthus emblica.

Swati Sharma,Vinita Hooda

doi:10.6026/97320630014008

Abstract

Catalase is an essential antioxidant enzyme that is well characterized from microbial and animal sources. The structure of plant catalase is unknown. Therefore, it is of interest to understand the functional and structural characteristics of catalase from an Indian gooseberry, Phyllanthus emblica (or Emblica officinalis). Hence, catalase from P. emblica was cloned in pUC18 plasmid, sequenced and submitted to GenBank with the accession numbers "MF979112" and "ATO98311.1". InterProScan showed that the coding sequence is monofunctional and haem-dependent catalase-like superfamily. Multiple sequence alignment (MSA) followed by phylogenetic analysis showed that the P. emblica catalase groups with soybean catalase. We further report the characteristics of structural model of the enzyme for functional characterization.

Highlights

Free radicals including reactive oxygen species (ROS) are regularly generated as byproducts of various metabolic reactions in a cell
Results and discussion: cDNA synthesis and sequencing: The agarose gel image presented in Figure 1A confirmed the isolation of RNA from P. emblica leaves (Figure 1A)
The cDNA synthesized from the purified RNA was analyzed on agarose gel and found to be approximately 500 bp long (Figure 1B)

Summary

Introduction

Free radicals including reactive oxygen species (ROS) are regularly generated as byproducts of various metabolic reactions in a cell. To counter the toxic effects of ROS, the eukaryotic cell produces various antioxidant enzymes including peroxidase, superoxide dismutase, polyphenol oxidase, catalase etc. Out of these enzymes, catalase is considered to be a highly active key antioxidant enzyme [2] that reduces oxidative stress by catalyzing the conversion of hydrogen peroxide to water and oxygen [3]. Catalase is considered to be a highly active key antioxidant enzyme [2] that reduces oxidative stress by catalyzing the conversion of hydrogen peroxide to water and oxygen [3] This enzyme shows a very high apparent Km in the range of 0.025 – 1722 mM and is not saturated with its substrate [4]. It is of interest to characterize catalase from P. emblica using structural models

Methods

Results

Conclusion