PCR-based screening and characterization of family-19 chitinase producing Streptomyces sp. from western Ghats forest soil

[...]five Streptomyces strains with potential chitinolytic activity were identified from the Agasthyaarkoodam region of Western Ghat. First little amount of soil from different fields were taken in petri plates and on them calcium carbonate was sprinkled, these was kept for drying for a period of 4-5 days to allow actinomycetes to sporulate. Primary Screening for Chitinase Activity: Streptomycetes isolates obtained from Glycerol asparagine medium were then screened for chitin hydrolysis by spoting on the centre of 1% colloidal chitin agar (cCa) (colloidalchitin1%; chitin (0.2%), MgSO4.7H2O (0.5%), K2HPO4(0.7%), KH2PO4 (0.3%), FeSO4.7H2O (0.01%), MnCl2(0.001%), NaCl (0.03%), Yeast extract (0.02%) and agar (2%), media at neutral pH and were incubated at 270C. the zone of clearance due to chitin hydrolysis was recorded up to 12 days. Selection ratio = Zone of clearance (mm) / Colony diameter (mm) DNA Extraction: Streptomyces isolates were cultured in YEME broth of composition (g/L) Yeast Extract- 3.0, Malt extract 3.0, sucrose-340.0, glucose-10.0, peptone-3.0 in 1000ml distilled water and pH-7.2 supplemented with 5mM MgCl2, and 0.5% (w/v) glycine.