Abstract
Problem statement: Although the diversity of marine Actinobacteria
have been studied and biotechnologically exploited throughout the world, the studies
on marine actinobacteria in Indian peninsula are largely unexplored. Of 9 maritime
states in India, only 4 states have been extensively studied for the diversity
of actinobacteria. Further, the studies on bioactive actinomycetes from saline
soil are very scanty. In the present study, we had taken an initiative to isolate
culturable halophilic actinomycetes and to screen the bioactive potential. Approach:
The marine sediment sample was collected at a depth of 400 cm at Marakkanam. The
strain was isolated using ISP No. 2 medium supplemented with 25% sea water. The
polyphasic taxonomy of the strain was evaluated by phenotypic, chemotaxonomic
and phylogenetic analysis. The 16S rRNA was sequenced and phylogenetic relationship
with the closest related species were studied. The growth conditions and the medium
had been optimized under shake-flask conditions by measuring the dry weight of
the mycelium. The isolate was subjected to fermentation and the crude extract
was screened for cytotoxic, hemolytic and antimicrobial activity. The cytotoxicity
was evaluated on HeLa cells by MTT assay, hemolytic assay on mouse erythrocytes
and the antimicrobial activity was determined by agar diffusion assay.
Results: Based on polyphasic taxonomy the species was identified as Saccharopolyspora
salina and belongs to the genus Saccharopolyspora. The 16S rRNA sequence of the
isolate showed 100% similarity with Saccharopolyspora salina. Based on the phylogenetic
and phenotypic evaluation the isolate was designated as Saccharopolyspora salina
VITSDK4. The growth was maximal in the designed production medium with the incubation
temperature of 28°C and pH of 7.4. It is a moderately halophilic species requires
9% NaCl concentration for optimal growth. The cytotoxicity on HeLa cells showed
the IC50 value of 26.2 µg mL-1 by MTT assay. The hemolytic activity on mouse
erythrocytes showed the EC50 value of 266 µg mL-1. The crude extract also
exhibited significant antagonistic activity against fungal pathogens Aspergillus
niger, Aspergillus fumigatus, Candida albicans and the Gram negative bacteria
Escherichia coli and Klebsiella pneumoniae. Only moderate activity was seen against
Gram positive bacteria Staphylococcus aureus and Bacillus subtilis. Conclusion/Recommendations:
Based on the results of our investigation, the isolated strain was identified
as Saccharopolyspora salina VITSDK4, which was moderately halophilic, produces
an extracellular bioactive metabolite, which inhibits the proliferation of HeLa
cells as well as antagonistic to fungal and bacterial pathogens. Further studies
on purification and characterization of the pure compound from the strain were
ongoing. The results of this study suggested that the marine actinomycetes from
the unexplored Indian coast could provide lead compounds of therapeutic value.