title: A metagenomic alkaline protease from saline habitat: cloning, over-expression and functional attributes.
abstract: Metagenomics has opened new horizon to unlock the biotechnological potential for novel enzymes. An alkaline protease gene was obtained from the total environmental DNA extracted from a saline habitat. After cloning and sequencing, it was identified that the protease gene related to uncultivable bacteria (HM219181). The protease was over expressed at 6h of induction with optimum induction at 1mM IPTG and 27°C. The purified enzyme was characterized with respect to various factors; temperature, pH, NaCl and chemical denaturant. The sequence analysis indicated a hydrophobic tendency of the protein, while the predicted 3D structure indicated the enzyme as a serine protease.
authors.affiliation: Department of Biosciences, Saurashtra University, Rajkot 360005, India.
authors.forename: Megha K
authors.initials: MK
authors.lastname: Purohit
authors.forename: Satya P
authors.initials: SP
authors.lastname: Singh
mesh_terms: D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology / Q000235:genetics; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D003001:Cloning, Molecular; D010450:Endopeptidases / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D004795:Enzyme Stability; D004926:Escherichia coli; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D054892:Metagenome; D056186:Metagenomics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010802:Phylogeny; D010957:Plasmids / Q000235:genetics / Q000302:isolation & purification; D011489:Protein Denaturation; D059748:Proteolysis; D055049:Salt Tolerance; D012965:Sodium Chloride / Q000737:chemistry; D012988:Soil Microbiology; D014508:Urea / Q000737:chemistry
chemical_list: D001426:Bacterial Proteins; D012965:Sodium Chloride; D014508:Urea; D010450:Endopeptidases; C087822:alkaline protease