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 a...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2022-10-01
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| Series: | FEBS Open Bio |
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| Online Access: | https://doi.org/10.1002/2211-5463.13457 |
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| author | Fabian Falkenberg Jade Rahba David Fischer Michael Bott Johannes Bongaerts Petra Siegert |
| author_facet | Fabian Falkenberg Jade Rahba David Fischer Michael Bott Johannes Bongaerts Petra Siegert |
| author_sort | Fabian Falkenberg |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-12T02:38:30Z |
| format | Article |
| id | doaj.art-4bacde5389144cbeb20ce3673aaf004b |
| institution | Directory Open Access Journal |
| issn | 2211-5463 |
| language | English |
| last_indexed | 2024-04-12T02:38:30Z |
| publishDate | 2022-10-01 |
| publisher | Wiley |
| record_format | Article |
| series | FEBS Open Bio |
| spelling | doaj.art-4bacde5389144cbeb20ce3673aaf004b2022-12-22T03:51:23ZengWileyFEBS Open Bio2211-54632022-10-0112101729174610.1002/2211-5463.13457Biochemical characterization of a novel oxidatively stable, halotolerant, and high‐alkaline subtilisin from Alkalihalobacillus okhensis Kh10‐101TFabian Falkenberg0Jade Rahba1David Fischer2Michael Bott3Johannes Bongaerts4Petra Siegert5Institute of Nano‐ and Biotechnologies Aachen University of Applied Sciences Jülich GermanyInstitute of Nano‐ and Biotechnologies Aachen University of Applied Sciences Jülich GermanyInstitute of Nano‐ and Biotechnologies Aachen University of Applied Sciences Jülich GermanyInstitute of Bio‐ and Geosciences IBG‐1: Biotechnology, Forschungszentrum Jülich GermanyInstitute of Nano‐ and Biotechnologies Aachen University of Applied Sciences Jülich GermanyInstitute of Nano‐ and Biotechnologies Aachen University of Applied Sciences Jülich GermanyHalophilic 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.https://doi.org/10.1002/2211-5463.13457Alkalihalobacillus okhensisdetergent proteasehalotolerant proteasehigh‐alkaline subtilisinoxidative stable protease |
| spellingShingle | Fabian Falkenberg Jade Rahba David Fischer Michael Bott Johannes Bongaerts Petra Siegert Biochemical characterization of a novel oxidatively stable, halotolerant, and high‐alkaline subtilisin from Alkalihalobacillus okhensis Kh10‐101T FEBS Open Bio Alkalihalobacillus okhensis detergent protease halotolerant protease high‐alkaline subtilisin oxidative stable protease |
| title | Biochemical characterization of a novel oxidatively stable, halotolerant, and high‐alkaline subtilisin from Alkalihalobacillus okhensis Kh10‐101T |
| title_full | Biochemical characterization of a novel oxidatively stable, halotolerant, and high‐alkaline subtilisin from Alkalihalobacillus okhensis Kh10‐101T |
| title_fullStr | Biochemical characterization of a novel oxidatively stable, halotolerant, and high‐alkaline subtilisin from Alkalihalobacillus okhensis Kh10‐101T |
| title_full_unstemmed | Biochemical characterization of a novel oxidatively stable, halotolerant, and high‐alkaline subtilisin from Alkalihalobacillus okhensis Kh10‐101T |
| title_short | Biochemical characterization of a novel oxidatively stable, halotolerant, and high‐alkaline subtilisin from Alkalihalobacillus okhensis Kh10‐101T |
| title_sort | biochemical characterization of a novel oxidatively stable halotolerant and high alkaline subtilisin from alkalihalobacillus okhensis kh10 101t |
| topic | Alkalihalobacillus okhensis detergent protease halotolerant protease high‐alkaline subtilisin oxidative stable protease |
| url | https://doi.org/10.1002/2211-5463.13457 |
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