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Biochemical characterization of a novel oxidatively stable, halotolerant, and high-alkaline subtilisin from Alkalihalobacillus okhensis Kh10-101T

  • Halophilic and halotolerant microorganisms represent a promising source of salt-tolerant enzymes suitable for various biotechnological applications where high salt concentrations would otherwise limit enzymatic activity. Considering the current growing enzyme market and the need for more efficient and new biocatalysts, the present study aimed at the characterization of a high-alkaline subtilisin from Alkalihalobacillus okhensis Kh10-101T. The protease gene was cloned and expressed in Bacillus subtilis DB104. The recombinant protease SPAO with 269 amino acids belongs to the subfamily of high-alkaline subtilisins. The biochemical characteristics of purified SPAO were analyzed in comparison with subtilisin Carlsberg, Savinase, and BPN'. SPAO, a monomer with a molecular mass of 27.1 kDa, was active over a wide range of pH 6.0–12.0 and temperature 20–80 °C, optimally at pH 9.0–9.5 and 55 °C. The protease is highly oxidatively stable to hydrogen peroxide and retained 58% of residual activity when incubated at 10 °C with 5% (v/v) H2O2 for 1 h while stimulated at 1% (v/v) H2O2. Furthermore, SPAO was very stable and active at NaCl concentrations up to 5.0 m. This study demonstrates the potential of SPAO for biotechnological applications in the future.

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Author:Fabian FalkenbergORCiD, Jade Rahba, David Fischer, Michael Bott, Johannes BongaertsORCiD, Petra SiegertORCiD
Parent Title (English):FEBS Open Bio
Place of publication:Hoboken, NJ
Document Type:Article
Year of Completion:2022
Date of the Publication (Server):2022/09/12
Tag:Alkalihalobacillus okhensis; detergent protease; halotolerant protease; high-alkaline subtilisin; oxidative stable protease
First Page:1729
Last Page:1746
Corresponding author: Petra Siegert
Institutes:FH Aachen / Fachbereich Chemie und Biotechnologie
FH Aachen / INB - Institut für Nano- und Biotechnologien
collections:Open Access / Gold
Licence (German):License LogoCreative Commons - Namensnennung