title: Production, Purification, and Biochemical Characterization of Thermostable Metallo-Protease from Novel Bacillus alkalitelluris TWI3 Isolated from Tannery Waste.
abstract: Protease enzymes in tannery industries have enormous applications. Seeking a potential candidate for efficient protease production has emerged in recent years. In our study, we sought to isolate proteolytic bacteria from tannery waste dumping site in Tamilnadu, India. Novel proteolytic Bacillus alkalitelluris TWI3 was isolated and tested for protease production. Maximum protease production was achieved using lactose and skim milk as a carbon and nitrogen source, respectively, and optimum growth temperature was found to be 40 °C at pH 8. Protease enzyme was purified using ammonium sulfate precipitation method and anion exchange chromatography. Diethylaminoethanol (DEAE) column chromatography and Sephadex G-100 chromatography yielded an overall 4.92-fold and 7.19-fold purification, respectively. The 42.6-kDa TWI3 protease was characterized as alkaline metallo-protease and stable up to 60 °C and pH 10. Ca(2+), Mn(2+), and Mg(2+) ions activated the protease, while Hg(2+), Cu(2+), Zn(2+), and Fe(2+) greatly inhibited it. Ethylenediaminetetraacetic acid (EDTA) inhibited TWI3 protease and was activated by Ca(2+), which confirmed that TWI3 protease is a metallo-protease. Moreover, this protease is capable of dehairing goat skin and also removed several cloth stains, which makes it more suitable for various biotechnological applications.
authors.affiliation: Department of Biology, Gandhigram Rural Institute-Deemed University, Gandhigram, 624302, Tamilnadu, India.|Biodiversity Research Center, Academia Sinica, Taipei, 11529, Taiwan.
authors.forename: Marimuthu
authors.initials: M
authors.lastname: Anandharaj
authors.affiliation: Department of Biology, Gandhigram Rural Institute-Deemed University, Gandhigram, 624302, Tamilnadu, India. balayogansivasankari@gmail.com.
authors.forename: Balayogan
authors.initials: B
authors.lastname: Sivasankari
authors.affiliation: Department of Biology, Gandhigram Rural Institute-Deemed University, Gandhigram, 624302, Tamilnadu, India.
authors.forename: Nagarajan
authors.initials: N
authors.lastname: Siddharthan
authors.affiliation: Department of Biology, Gandhigram Rural Institute-Deemed University, Gandhigram, 624302, Tamilnadu, India.
authors.forename: Rizwana Parveen
authors.initials: RP
authors.lastname: Rani
authors.affiliation: Department of Biology, Gandhigram Rural Institute-Deemed University, Gandhigram, 624302, Tamilnadu, India.|Department of Biotechnology, Indian Institute of Technology Madras, Chennai, 600036, Tamilnadu, India.
authors.forename: Subramaniyan
authors.initials: S
authors.lastname: Sivakumar
mesh_terms: D001407:Bacillus / Q000201:enzymology*; D004795:Enzyme Stability; D007194:India; D007220:Industrial Waste; D007700:Kinetics; D045726:Metalloproteases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D011480:Protease Inhibitors / Q000494:pharmacology; D059748:Proteolysis; D013696:Temperature
chemical_list: D007220:Industrial Waste; D011480:Protease Inhibitors; D045726:Metalloproteases
keywords: Bacillus alkalitelluris; Dehairing; Destaining; Protease; Tannery