Purification and characterization of an extracellular thermo-alkali stable, metal tolerant chitinase from Streptomyces chilikensis RC1830 isolated from a brackish water lake sediment

Lopamudra Ray; Ananta Narayan Panda; Samir Ranjan Mishra; Ajit Kumar Pattanaik; Tapan Kumar Adhya; Mrutyunjay Suar; Vishakha Raina

doi:10.1016/j.btre.2019.e00311

Purification and characterization of an extracellular thermo-alkali stable, metal tolerant chitinase from Streptomyces chilikensis RC1830 isolated from a brackish water lake sediment

Biotechnol Rep (Amst). 2019 Jan 29:21:e00311. doi: 10.1016/j.btre.2019.e00311. eCollection 2019 Mar.

Authors

Lopamudra Ray^{1

2}, Ananta Narayan Panda¹, Samir Ranjan Mishra¹, Ajit Kumar Pattanaik³, Tapan Kumar Adhya¹, Mrutyunjay Suar¹, Vishakha Raina¹

Affiliations

¹ School of Biotechnology, KIIT University, Bhubaneswar, Odisha, 751024, India.
² School of Law, KIIT University, Bhubaneswar, Odisha, 751024, India.
³ Wetland Research and Training Centre, Chilika Development Authority, Barkul, Balugaon, 751014, Odisha, India.

Abstract

An extracellular thermo-alkali stable chitinase was obtained from Streptomyces chilikensis RC1830, a novel actinobacterial strain isolated from the sediments of Chilika lake, India. Purification of the enzyme was carried out by concentrating the enzyme with centrifugal device followed by chromatographic separation by DEAE Sepharose ion exchange resin.The molecular weight of the enzyme was 10.5 kDa as determined by SDS-PAGE. The optimum pH and temperature for the partially purified chitinase was pH 7 and 60 °C. The chitinase showed 40% activity at pH 11 after 24 h exposure at room temperature. The chitinase exhibited Km and Vmax values are 0.02 mM and 3.184 mol/min/mg of enzyme respectively. The 6 residue N-terminal sequence of the enzyme was not found similar to any of the reported chitinase enzyme. Based on the SDS PAGE, zymogram analysis, activity assays and other characteristics, it is proposed that the purified enzyme from S.chilikensis RC1830 is a chitinase.

Keywords: Chio oligosaccharides; Chitinase; Halo tolerant; Streptomyces.