Production of Fibrinolytic Protease from a Halobacterium Bacillus licheniformis VITLMS Isolated from Marine Sponges of Rameshwaram Coast, India

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Author(s): Lalith Kumar .CK, Merlyn Keziah.S, Hemalatha. M, Mohanapriya.A, Mohanasrinivasan.V, Subathra Devi.C*

Journal Name: Current Bioactive Compounds

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Background: Marine bacteria serve as excellent sources of therapeutic enzymes, metabolites and natural products, which possess novel therapeutic properties. Increasing death rates due to cardiovascular diseases urges for cost effective production of fibrinolytic enzyme.

Methods: Marine sponge samples were screened for potent fibrinolytic producing bacteria. Primary screening was done for protease production, clot lysis activity. Secondary screening was done for casein plasminogen activity and fibrinolytic activity. The strain which had potent fibrinolytic activity among them was further subjected to morphological, biochemical and molecular characterization. Media optimization was carried to enhance enzyme production. The enzyme produced was subjected to purification using ammonium sulfate precipitation, gel filtration and characterized using HPLC and FTIR analysis

Results: Sponge was identified to be Desmapsamma anchorata. Thirteen bacterial isolates were isolated from the sponge sample. . The 16S rRNA sequencing revealed that the potential strain had 99% similarity with Bacillus licheniformis. Amongst the isolates most were found to be morphologically identical to Bacillus genus. Gram’s staining and SEM analysis of the potent isolate was performed to identify the spore formation and rod shaped morphology of the bacteria. The optimal temperature and pH for production of the enzyme was 37 °C and 8 respectively. The carbon source maltose and nitrogen sources were malt extract and yeast extract were found to be optimal. The optimum incubation time was found to be both 4 to 5 days. The crude supernatant was purified with ammonium sulfate precipitation and gel filtration chromatography. The retention time of 11.3 min and presence of functional groups show the purity of the enzyme. The partially purified enzyme showed 96.4 % clot lysis in artificial clot lysis activity. Conclusion: Although the secretion of fibrinolytic enzymes from Bacillus species is not new, based on our investigation there are no reports regarding Bacillus licheniformis being isolated from marine sponges. However, there are reports of Bacillus licheniformis secreting fibrinolytic enzymes isolated from fermented food samples This study identifies marine environment as a potential source of new exploration for drug discovery.

Keywords: Desmapsamma anchorata, Bacillus licheniformis, Clot lysis, fibrinolytic, Halobacterium.

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(E-pub Ahead of Print)
DOI: 10.2174/1573407216666200330112127
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