title: Screening and characterization of protease producing actinomycetes from marine saltern.
abstract: In the course of systematic screening program for bioactive actinomycetes, an alkaline protease producing halophilic strain Actinopolyspora sp. VITSDK2 was isolated from marine saltern, Southern India. The strain was identified as Actinopolyspora based on its phenotypic and phylogenetic characters. The protease was partially purified using ammonium sulfate precipitation and subsequently by DEAE cellulose column chromatography. The enzyme was further purified using HPLC and the molecular weight was found to be 22 kDa as determined by SDS-PAGE analysis. The purified protease exhibited pH stability in a wide range of 4-12 with optimum at 10.0. The enzyme was found to be stable between 25 and 80 °C and displayed a maximum activity at 60 °C. The enzyme activity was increased marginally in presence of Mn(2+) , Mg(2+) , and Ca(2+) and decreased in presence of Cu(2+) . PMSF and DFP completely inhibited the activity suggesting it belongs to serine protease. Further, the proteolytic activity was abolished in presence of N-tosyl-L-lysine chloromethyl ketone suggesting this might be chymotrypsin-like serine protease. The protease was 96% active when kept for 10 days at room temperature. The results indicate that the enzyme belong to chymotrypsin-like serine protease exhibiting both pH and thermostability, which can be used for various applications in industries.
authors.affiliation: Marine Biotechnology and Biomedicine Laboratory, School of Biosciences and Technology, VIT University, Vellore, India.
authors.forename: Krish
authors.initials: K
authors.lastname: Suthindhiran
authors.forename: Mangalam Achuthananda
authors.initials: MA
authors.lastname: Jayasri
authors.forename: Dipa
authors.initials: D
authors.lastname: Dipali
authors.forename: Apurva
authors.initials: A
authors.lastname: Prasar
mesh_terms: D039903:Actinobacteria / Q000145:classification / Q000201:enzymology* / Q000302:isolation & purification* / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D010450:Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D004795:Enzyme Stability; D019015:Geologic Sediments / Q000382:microbiology*; D007194:India; D054712:Salinity; D057057:Serine Proteases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D010450:Endopeptidases; D057057:Serine Proteases; C087822:alkaline protease
keywords: Actinopolyspora; Alkaliphilic; Halophilic; Marine; Protease