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New robust subtilisins from halotolerant and halophilic Bacillaceae
Fabian Falkenberg1, Leonie Voß1, Michael Bott2
1Institute of Nano- and Biotechnologies, Aachen University of Applied Sciences, 52428, Jülich, Germany.
Applied Microbiology and Biotechnology
|May 9, 2023
Summary
Researchers characterized four novel subtilisins from halophilic and halotolerant bacteria. These enzymes exhibit broad pH and salt tolerance, showing potential for industrial biocatalyst applications.
Area of Science:
- Biochemistry
- Enzymology
- Microbiology
Background:
- The expanding enzyme market demands novel biocatalysts.
- Halophilic and halotolerant Bacillaceae are promising sources for uncharacterized enzymes.
- Subtilisins are a class of proteases with diverse industrial applications.
Purpose of the Study:
- To characterize four new subtilisins from halotolerant and halophilic microorganisms.
- To evaluate the biochemical properties and potential applications of these novel enzymes.
Main Methods:
- Data mining of halophilic/halotolerant Bacillaceae genomes.
- Cloning and expression of protease genes in Bacillus subtilis.
- Purification using ethanol precipitation, desalting, and ion-exchange chromatography.
- Biochemical characterization of enzyme activity, stability, and optimal conditions.
Main Results:
- Four subtilisins (SPPM, SPAH, SPMI, SPLA) were successfully cloned, expressed, and purified.
- Enzyme activity was observed across a wide pH range (5.0-12.0) with optima at pH 9.0-10.0.
- Optimal temperatures ranged from 50 °C to 70 °C.
- Proteases demonstrated high stability in 5% SDS and activity at 5 M NaCl.
Conclusions:
- Halophilic and halotolerant Bacillaceae are a valuable source for novel subtilisins.
- The characterized subtilisins exhibit robust biochemical properties suitable for industrial use.
- These novel proteases hold significant potential for future biotechnological applications.
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