Production, Purification, and Biochemical Characterization of Thermostable Metallo-Protease from Novel Bacillus alkalitelluris TWI3 Isolated from Tannery Waste

被引:32
作者
Anandharaj, Marimuthu [1 ,2 ]
Sivasankari, Balayogan [1 ]
Siddharthan, Nagarajan [1 ]
Rani, Rizwana Parveen [1 ]
Sivakumar, Subramaniyan [1 ,3 ]
机构
[1] Deemed Univ, Gandhigram Rural Inst, Dept Biol, Gandhigram 624302, Tamil Nadu, India
[2] Acad Sinica, Biodivers Res Ctr, Taipei 11529, Taiwan
[3] Indian Inst Technol, Dept Biotechnol, Madras 600036, Tamil Nadu, India
关键词
Protease; Bacillus alkalitelluris; Tannery; Dehairing; Destaining; ALKALINE PROTEASE; SERINE-PROTEASE; OPTIMIZATION; EXTRACTION;
D O I
10.1007/s12010-015-1974-7
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 A degrees 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 A degrees C and pH 10. Ca2+, Mn2+, and Mg2+ ions activated the protease, while Hg2+, Cu2+, Zn2+, and Fe2+ greatly inhibited it. Ethylenediaminetetraacetic acid (EDTA) inhibited TWI3 protease and was activated by Ca2+, 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.
引用
收藏
页码:1666 / 1686
页数:21
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