Single-site substitutions improve cold activity and increase thermostability of the dehairing alkaline protease (DHAP). Issue 12 (1st December 2016)

• Record Type: Journal Article
• Authors: Zhao, Hong-Yan; Wu, Li-Ying; Liu, Gang; Feng, Hong
• Is Part Of: Bioscience, biotechnology, and biochemistry. Volume 80:Issue 12(2016)
• Page Start: 2480
• Page End: 2485
• Publication Date: 2016-12-01
• Subjects: alkaline protease -- Bacillus pumilus -- cold activity -- site-directed mutagenesis -- thermostability; Biotechnology -- Periodicals; Biochemistry -- Periodicals; 660.6
• DOI: 10.1080/09168451.2016.1230005
• Languages: English
• ISSNs: 0916-8451

Abstract

Abstract: To engineer dehairing alkaline protease (DHAP) variants to improve cold activity and increase thermostability so these variants are suitable for the leather processing industry. Based on previous studies with bacterial alkaline proteases, double-site mutations (W106K/V149I and W106K/M124L) were introduced into the DHAP from Bacillus pumilus . Compared with the wild-type DHAP hydrolytic activity, the double-site variant W106K/V149I showed an increase in specific hydrolytic activity at 15 °C by 2.3-fold toward casein in terms of hydrolytic rate and 2.7-fold toward the synthetic peptide AAPF- p N by means of k cat / K m value. The thermostability of the variant (W106K/V149I) was improved with the half-life at 60 and 70 °C increased by 2.7- and 5.0-fold, respectively, when compared with the thermostability of the wild-type DHAP. Conclusively, an increase in the cold activity and thermostability of a bacterial alkaline protease was achieved by protein engineering. Graphical abstract: : This article mainly describes the protease of DHAP can inprove thermostability and activity through the site combination.