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Biochemical characterisation of a novel broad pH spectrum subtilisin from Fictibacillus arsenicus DSM 15822ᵀ

  • Subtilisins from microbial sources, especially from the Bacillaceae family, are of particular interest for biotechnological applications and serve the currently growing enzyme market as efficient and novel biocatalysts. Biotechnological applications include use in detergents, cosmetics, leather processing, wastewater treatment and pharmaceuticals. To identify a possible candidate for the enzyme market, here we cloned the gene of the subtilisin SPFA from Fictibacillus arsenicus DSM 15822ᵀ (obtained through a data mining-based search) and expressed it in Bacillus subtilis DB104. After production and purification, the protease showed a molecular mass of 27.57 kDa and a pI of 5.8. SPFA displayed hydrolytic activity at a temperature optimum of 80 °C and a very broad pH optimum between 8.5 and 11.5, with high activity up to pH 12.5. SPFA displayed no NaCl dependence but a high NaCl tolerance, with decreasing activity up to concentrations of 5 m NaCl. The stability enhanced with increasing NaCl concentration. Based on its substrate preference for 10 synthetic peptide 4-nitroanilide substrates with three or four amino acids and its phylogenetic classification, SPFA can be assigned to the subgroup of true subtilisins. Moreover, SPFA exhibited high tolerance to 5% (w/v) SDS and 5% H₂O₂ (v/v). The biochemical properties of SPFA, especially its tolerance of remarkably high pH, SDS and H₂O₂, suggest it has potential for biotechnological applications.

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Author:Fabian FalkenbergORCiD, Sophie Kohn, Michael Bott, Johannes BongaertsORCiD, Petra SiegertORCiD
Parent Title (English):FEBS Open Bio
Place of publication:Hoboken, NJ
Document Type:Article
Year of Completion:2023
Date of first Publication:2023/08/30
Date of the Publication (Server):2023/09/11
Tag:Bacillaceae; Biotechnological application; Broad pH spectrum; Subtilases; Subtilisin
First Page:2035
Last Page:2046
Corresponding author: Petra Siegert
Institutes:FH Aachen / Fachbereich Chemie und Biotechnologie
FH Aachen / INB - Institut für Nano- und Biotechnologien
collections:Verlag / Wiley
Open Access / Gold
Licence (German):License LogoCreative Commons - Namensnennung