MOLECULAR CHARACTERIZATION OF CHITINOLYTIC STRAINS ISOLATED FROM TERRESTRIAL HABITATS

Author(s): Monisha Khanna Kapur*, Payal Das, Prateek Kumar, Munendra Kumar and Renu Solanki

Abstract: Three actinomycete strains producing extracellular chitinase, designated as colony 130, 194 and 184 were isolated from soil samples collected from NTPC chemical plant, (Faridabad) Haryana, landfill site (Sarai Kale Khan) Delhi and sugar plant Mandi Dhanaura, U.P respectively. During primary screening diameter of zones of hydrolysis were in the range of 8 to 16 mm as reported in our previous paper Das et al., 2017. Isolates 130, 194, 184 and Streptomyces albidoflavus (NRRLB 16746) as positive control were subsequently subjected to secondary level of screening and purification. Activity of enzyme in crude extracts was in the range of 7.16 -14.12 IU/ml and in partially purified samples in the range 12.1 to 23.10 IU/ml as reported in our earlier studies [14]. Comparison and analysis of the 16S rRNA gene sequence revealed that colonies 130, 194 and 184 belonged to the genus Streptomyces. Colony 130 (Accession no: KY357309) had 97.23% sequence similarity with Streptomyces calvus ISP 5010T (AY999780), colony 194 (Accession no: KJ934595) had 96.20% similarity with Streptomyces albidoflavus DSM 40455T (Z76676) and colony 184 (KY357314) showed 98.8% similarity with Streptomyces enissocaesilis NRRL B-16365T (DQ026641). Random amplified polymorphic DNA (RAPD) was assayed to determine the genetic diversity of colonies 130 and 194 with reference strain, Streptomyces albidoflavus. Colony 130 showed around 80-90% interspecific variation, the colony has different genetic origin in comparison to the reference strain. On the other hand, colonies 194 and 184 showed around 50-60% and 20% variation respectively and hence have almost same genetic origin as S. albidoflavus.

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