IJSTE - International Journal of Science Technology & Engineering | Volume 3 | Issue 02 | August 2016 ISSN (online): 2349-784X All rights reserved by www.ijste.org 169 Enhancement and Detection of Chitinase Enzyme of Fungus for the Eradication of Pests Priya D Sneka S UG Student UG Student Department of Biotechnology Department of Biotechnology Adhiyamaan College of Engineering (Autonomous) Hosur, Tamil Nadu Adhiyamaan College of Engineering (Autonomous) Hosur, Tamil Nadu Ramesh Babu N G Karthikeyan S Professor & Head Assistant Professor Department of Biotechnology Department of Biotechnology Adhiyamaan College of Engineering (Autonomous) Hosur, Tamil Nadu Adhiyamaan College of Engineering (Autonomous) Hosur, Tamil Nadu Abstract Chitinase is an enzyme having the ability to degrade chitin to a low molecular weight chitooligomers which have a wide range of applications. In the present study, chitinase was produced from the entomopathogenic fungi such as Pochonia chlamydosporia, Beauveria bassiana, Metarhizium anisopliae and Trichoderma harzianum using chitin as a carbon source. The enzyme was partially purified by ammonium sulphate precipitation and the activity was detected using colorimetric method. The enzyme showed the maximum activity in Trichoderma harzianum among the four fungal species at 40°C and pH 6.5. This enzyme could be used directly to control of pathogenic fungi and various pests in the egg stage itself. Keywords: Chitinase, Chitosan, Pochonia Chlamydosporia, Beauveria Bassiana, Metarhizium Anisopliae, Trichoderma Harzianum ________________________________________________________________________________________________________ I. INTRODUCTION Dendrobium is a diverse genus of orchids which is easily discernable from other plant species in the wild by its very distinguishable and unique flower, which comes in many colours, size. It is very expensive. This plant is attacked by the pest - Dendrobium blossom midge, Contanaria maculipennis and the pest epicuticle is made up of chitin; it acts as a physical barrier to the pest. There is no effective biological or chemical control against blossom midge [1]. Chitin is a ȕ -1, 4 N-acetyl glycosamine polymer which is 2-acetamido-2-deoxy-D-glucose linked by ȕ-(1-4) glycosidic bonds and is the second highest occurring biopolymer [β]. It exists in nature in three different forms α-chitin, ȕ-chitin and Ȗ-chitin [3] and it is found in various species such as marine vertebrates, insects, fungi and algae [4], it is mainly obtained from marine vertebrates for commercial purpose and more than 80,000 metric tons of chitin is produced per year from marine waste [5]. Alkaline hydrolysis of chitin chains give rise to chitosan, which is relatively nontoxic and has antimicrobial properties. Chitinases (E.C.3.2.2.14) are glycosyl hydrolases, having the ability to degrade chitin as a renewable source [4]. It is classified into two main classes, endochitinase and exochitinase. The endochitinase randomly splits chitin at internal sides, thereby forming the di-cetylchitotriose and low molecular mass multimers of GlcNAc. The exochitinase cleave chitin from its nonreducing end, releasing dimers (GlcNAC)2 [6]. Chitinase plays an effective role in the controlling of pests and insects that can be an alternative to chemical pesticides and it is present in a wide range of organisms such as bacteria, virus, protozoa, fungi, plants and animals to reshape their own chitin [7]. Among all the biological control agents, entamopathogenic fungi play a major role in controlling pest due to its broad host range. The entamopathogenic fungi like Pochonia chlamydosporia, Beauveria bassiana, Metarhizium anisopliae and Trichoderma harzianum are more efficient for the control of pathogens by the production of chitinase which are secreted by fungi in liquid culture supplemented with chitin as a carbon source [8]. The enzyme could either be used directly in the biological control or through gene manipulation [9]. Apart from application of chitinase as biopesticides, the chitinase have also been used for production of single cell proteins [10], and for the production of fungal protoplasts [11]. The present study was aimed to detect chitinase enzyme in entamopathogenic fungi for the eradication of pests employing chitin as a carbon source.