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http://dx.doi.org/10.4014/mbl.1606.06002

Purification and Characterization of an Alkaline Protease Produced by Alkalophilic Bacillus sp. DK1122  

Lee, Hyungjae (Department of Food Engineering, Dankook University)
Yoo, Ji-Seung (Department of Food Engineering, Dankook University)
Bai, Dong-Hoon (Department of Food Engineering, Dankook University)
Publication Information
Microbiology and Biotechnology Letters / v.44, no.3, 2016 , pp. 333-340 More about this Journal
Abstract
An alkaline protease was purified and characterized from an alkalophilic microorganism, Bacillus sp. DK1122, isolated from soil in central Korea. The optimum temperature and pH for the growth of the producer strain were 40℃ and pH 9.0, respectively. The protease was produced aerobically at 40℃ after 24 h incubation in modified Horikoshi I medium (pH 9.0) containing 0.5% (w/v) glucose, 0.8% (w/v) yeast extract, 0.5% (w/v) polypeptone, 0.1% (w/v) K2HPO4, 0.02% (w/v) MgSO4·7H2O, 1% (w/v) Na2CO3, and 3% (w/v) NaCl. The alkaline protease was purified by 70% ammonium sulfate precipitation of the culture supernatant of Bacillus sp. DK1122, followed by CM-Sepharose chromatography. The molecular weight of the enzyme was estimated to be 27 kDa on the basis of SDS-PAGE. The optimum temperature and pH for the protease activity were 60℃ and pH 9.0, respectively. Addition of CaCl2 increased the thermal stability of the purified protease, where 90% of protease activity was retained at 60℃ for up to 3 h. Consequently, it is expected that the alkaline protease from this study, exhibiting stability at pH 7–9 and 60℃, may be promising for application in the food and detergent industries.
Keywords
Alkaline protease; alkalophilic; Bacillus; purification; characterization;
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