TY - JOUR A1 - Falkenberg, Fabian A1 - Rahba, Jade A1 - Fischer, David A1 - Bott, Michael A1 - Bongaerts, Johannes A1 - Siegert, Petra T1 - Biochemical characterization of a novel oxidatively stable, halotolerant, and high-alkaline subtilisin from Alkalihalobacillus okhensis Kh10-101T JF - FEBS Open Bio N2 - 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. KW - Alkalihalobacillus okhensis KW - detergent protease KW - halotolerant protease KW - high-alkaline subtilisin KW - oxidative stable protease Y1 - 2022 U6 - http://dx.doi.org/10.1002/2211-5463.13457 SN - 2211-5463 N1 - Corresponding author: Petra Siegert VL - 12 IS - 10 SP - 1729 EP - 1746 PB - Wiley CY - Hoboken, NJ ER - TY - JOUR A1 - Falkenberg, Fabian A1 - Voß, Leonie A1 - Bott, Michael A1 - Bongaerts, Johannes A1 - Siegert, Petra T1 - New robust subtilisins from halotolerant and halophilic Bacillaceae JF - Applied Microbiology and Biotechnology N2 - 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. KW - Biotechnological application KW - Bacillaceae KW - Subtilisin KW - Subtilases KW - Halotolerant protease Y1 - 2023 U6 - http://dx.doi.org/10.1007/s00253-023-12553-w SN - 1432-0614 N1 - Corresponding author: Petra Siegert VL - 107 SP - 3939 EP - 3954 PB - Springer Nature CY - Berlin ER - TY - JOUR A1 - Feng, Yong-Qing A1 - Seibler, Jost A1 - Alami, Raouf A1 - Eisen, Andrew A1 - Westerman, Karen A. A1 - Leboulch, Philippe A1 - Fiering, Steven A1 - Bouhassira, Eric E. T1 - Site-specific chromosomal integration in mammalian cells: highly efficient CRE recombinase-mediated cassette exchange JF - Journal of Molecular Biology Y1 - 1999 U6 - http://dx.doi.org/10.1006/jmbi.1999.3113 SN - 0022-2836 VL - 292 IS - 4 SP - 779 EP - 785 ER - TY - JOUR A1 - Feuerriegel, Uwe T1 - Trennen und Trocknen. FH Aachen verfügt über einen Zentrifugentrockner im Technikumsmaßstab JF - Lebensmitteltechnik. 33 (2001), H. 7-8 Y1 - 2001 SN - 0047-4290 SP - 22 EP - 23 ER - TY - BOOK A1 - Feuerriegel, Uwe T1 - Zur Desaktivierung eines Desaktivierung eines Palladium-Trägerkatalysators durch Schwefel¬wasserstoff bei der katalytischen Methanoxidation Y1 - 1993 N1 - Dissertation PB - Univ.-Gesamthochsch. CY - Kassel ER - TY - JOUR A1 - Feuerriegel, Uwe T1 - Energetische Verwertung von Kunststoffabfällen JF - Chemie und Technologie makromolekularer Stoffe :, 16. Kolloquium. / Fachhochschule / Fachbereich Chemieingenieurwesen. 20. November 1998 an der FH Aachen, Fachbereich Chemieingenieurwesen Y1 - 1998 N1 - FH-Texte. 69; In Jülich unter der Signatur: 61 ZOW 405-16 SP - 169 EP - 189 PB - Fachhochschule Aachen CY - Aachen ER - TY - BOOK A1 - Feuerriegel, Uwe T1 - Verfahrenstechnik mit EXCEL: Verfahrenstechnische Berechnungen effektiv durchführen und professionell dokumentieren Y1 - 2016 SN - 978-3-658-02902-9 U6 - http://dx.doi.org/10.1007/978-3-658-02903-6 N1 - 152 Abbildungen, 94 Abbildungen in Farbe Gedruckt in der Bibliothek unter der Signatur: 21 ZNK 11 PB - Springer Fachmedien CY - Wiesbaden ER - TY - JOUR A1 - Feuerriegel, Uwe A1 - Hoffmann, Ulrich A1 - Walenzik, C. A1 - Kögel, J. T1 - Für Scale-up´s und Betriebsmuster, Projekt Trocknungstechnik an der FH Aachen JF - CITplus : das Praxismagazin für Verfahrenstechnik und Apparatebau. 4 (2001) Y1 - 2001 SN - 1436-2597 SP - 24 EP - 25 ER - TY - JOUR A1 - Feuerriegel, Uwe A1 - Klose, W. T1 - Reaktionskinetik der katalytischen Methan-Oxidation und experimentelle Untersuchungen zur Desaktivierung eines Palladium-Trägerkatalysators durch Schwefelwasserstoff. Klose, W. ; Feuerriegel, U. JF - Chemie - Ingenieur - Technik. 65 (1993), H. 10 Y1 - 1993 SN - 0009-286X N1 - engl. Titel: Reaction Kinetics of Catalytic Methane Oxidation and Deactivation of a Palladium Catalyst by Hydrogen Sulfide During Catalytic Oxidation of Methane SP - 1242 EP - 1243 ER - TY - JOUR A1 - Feuerriegel, Uwe A1 - Klose, W. T1 - Untersuchungen zum Mechanismus der Desaktivierung eines Palladium-Trägerkatalysators durch Vergiftung mit Schwefelwasserstoff JF - Chemie - Ingenieur - Technik. 70 (1998), H. 4 Y1 - 1998 SN - 0009-286X N1 - Artikel in Englisch unter Chemical engineering & technology. 21(1998)7. S. 562-564 SP - 449 EP - 452 ER -