Halophilic and Halotolerant Actinomycetes of Sambhar Salt Lake, India: Screening and Optimization of Cellulolytic Activity

Abstract

Actinomycetes are Gram-positive filamentous bacteria well known for the production of bioactive compounds. Recently, many halophilic habitats have been explored for isolation of actinomycetes that exhibit biotechnological potentials. In this investigation, a saline habitat of Rajasthan, Sambhar Salt Lake (SSL) was selected to study the actinomycetes population and Carboxy Methyl Cellulase (CMCase) production by native isolates. A total of sixteen actinomycete isolates, halotolerant and moderately halophilic, were obtained using culture-dependent methods and characterized morphologically and biochemically. They were identified as members of Streptomyces, Nocardiopsis, Pseudonocardia, Saccharospolyspora, and Microbispora. Streptomyces was the most dominating genus, followed by Nocardiopsis. Agar plate assay was used for screening the isolates for CMCase production. Thirteen were found to produce the enzyme, apparent by hydrolysis observed on media plates. The highest relative activity of 22.04 was shown by isolate SSL 14 identified as Nocardiopsis sp. by 16S rDNA sequencing studies and thus selected for further optimization studies. Maximum enzyme (1.08 ± 0.09 U/ml) was produced using medium containing Carboxy Methyl Cellulose (Carbon source) and yeast extract (nitrogen source) at 12% NaCl and pH 9.0, incubated at 30 °C for 96 h. Maximum CMCase production at high salt concentration and pH suggests that Nocardiopsis SSL 14 can be used for industrial processes that operate under excessive saline and alkaline conditions.