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New robust subtilisins from halotolerant and halophilic Bacillaceae

  • The aim of the present study was the characterisation of three true subtilisins and one phylogenetically intermediate subtilisin from halotolerant and halophilic microorganisms. Considering the currently growing enzyme market for efficient and novel biocatalysts, data mining is a promising source for novel, as yet uncharacterised enzymes, especially from halophilic or halotolerant Bacillaceae, which offer great potential to meet industrial needs. Both halophilic bacteria Pontibacillus marinus DSM 16465ᵀ and Alkalibacillus haloalkaliphilus DSM 5271ᵀ and both halotolerant bacteria Metabacillus indicus DSM 16189 and Litchfieldia alkalitelluris DSM 16976ᵀ served as a source for the four new subtilisins SPPM, SPAH, SPMI and SPLA. The protease genes were cloned and expressed in Bacillus subtilis DB104. Purification to apparent homogeneity was achieved by ethanol precipitation, desalting and ion-exchange chromatography. Enzyme activity could be observed between pH 5.0–12.0 with an optimum for SPPM, SPMI and SPLA around pH 9.0 and for SPAH at pH 10.0. The optimal temperature for SPMI and SPLA was 70 °C and for SPPM and SPAH 55 °C and 50 °C, respectively. All proteases showed high stability towards 5% (w/v) SDS and were active even at NaCl concentrations of 5 M. The four proteases demonstrate potential for future biotechnological applications.

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Author:Fabian FalkenbergORCiD, Leonie Voß, Michael Bott, Johannes BongaertsORCiD, Petra SiegertORCiD
Parent Title (English):Applied Microbiology and Biotechnology
Publisher:Springer Nature
Place of publication:Berlin
Document Type:Article
Year of Completion:2023
Date of first Publication:2023/05/09
Date of the Publication (Server):2023/05/10
Tag:Bacillaceae; Biotechnological application; Halotolerant protease; Subtilases; Subtilisin
First Page:3939
Last Page:3954
Corresponding author: Petra Siegert
Institutes:FH Aachen / Fachbereich Chemie und Biotechnologie
FH Aachen / INB - Institut für Nano- und Biotechnologien
collections:Verlag / Springer Nature
Open Access / Hybrid
Licence (German):License LogoCreative Commons - Namensnennung