Molecular study on the extracellular chitinolytic activity and antifungal activity of bacillus subtilis ts22

An extracellular chitinase producing bacterium Bacillus subtilis TS22 was isolated from shrimp shell waste. It was identified by as gram positive rod shaped bacteria, Indole, Citrate, Oxidase, Catalase, Nitrate positive and Methyl Red, Vogeus Proskauer negative. The carbohydrate fermentation occurs by producing acid but not gas in Glucose, Maltose and Sucrose. The genomic size of Bacillus subtilis TS22 was 870bp confirmed with the help of 16s rRNA primer viz forward primer F5- AGAGTTTGATCMTGGCTCAG-3| and reverse primer 5|AAGGAGGTGWTCCARCC-3|.The obtained gene sequence were submitted and deposited to NCBI under accession number JQ 727436. When plotted for phylogenetic tree showed 98% similarities with Bacillus species. The Bacillus subtilis TS22 produced the extracellular chitinase having 127U/ml of chitinolytic activity. The molecular weight of chitinase was 50KDa in 12% sodium dodecyl sulphate polyacrylamide gel electrophoresis. The Bacillus subtilis TS22 showed antifungal activity against three fungal pathogens such as Phytophthora parasitica, Alternaria solani and Pythium aphanidermatum.