Optimization of mycolytic enzymes (Chitinase, <span style="font-size:15.0pt;font-family:Symbol;mso-bidi-font-family:Symbol"><img src='/image/spc_char/beta.gif' border=0>1,3-Glucanase and <span style="font-size:15.0pt;font-family:"Times New Roman";mso-fareast-font-family: "Times New Roman";mso-ansi-language:EN-IN;mso-fareast-language:EN-IN; mso-bidi-language:AR-SA">Cellulase) production by <i>Bacillus subtilis</i>, a potential biocontrol agent using one-factor approach</span></span>

Abstract

This study presents one-factor approach for optimization of growth and physical parameters for increasing production of three mycolytic enzymes <i>viz. </i>chitinase, <span style="font-size: 9.0pt;font-family:Symbol;mso-bidi-font-family:Symbol"><img src='/image/spc_char/beta.gif' border=0>-1,3-glucanase and <span style="font-size:9.0pt; font-family:Symbol;mso-bidi-font-family:Symbol"><img src='/image/spc_char/beta.gif' border=0>1,4-glucanase (cellulase) by <i>B.subtilis</i>, a potent biocontrol agent. Maximum enzyme activity of all three enzymes was observed in the following physical parameters: pH 7.0, temperature of 30°C, and incubation period of 72h. Optimum nutritional parameters were found to be as follows: Carboxymethyl cellulose (CMC, 10g/L); Corn steep liquor (CSL, 1g/L) or KNO₃ (1 g/L), CaCl₂ (5mM)and Triton X 100 (0.1% v/v). Using one factor approach, production of all three enzymes ignificantly increased for chitinase (5.49 folds); â-1,3-glucanase (7.39 folds) and cellulase (1.91 folds) respectively as compared to the un-optimized media containing Nutrient broth. The crude mycolytic enzymes had activity in acidic pH (4 - 6) and temperature (50 - 60°C) range with optima at pH 6.0 and 55°C, respectively. However, chitinase showed two pH optima at pH 6 and 11 indicating the possibility of having two isoforms. The enzymes showed varied response to metal ions. Identification of essential nutrients affecting co-production of three enzymes by a single strain would help to formulate a suitable medium for their concerted production. Moreover, low cost cellulosic materials can be used as carbon source for these enzymes production by the isolate for industrial and agricultural applications.

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