TY  - JOUR
A1  - Falkenberg, Fabian
A1  - Kohn, Sophie
A1  - Bott, Michael
A1  - Bongaerts, Johannes
A1  - Siegert, Petra
T1  - Biochemical characterisation of a novel broad pH spectrum subtilisin from Fictibacillus arsenicus DSM 15822ᵀ
JF  - FEBS Open Bio
N2  - 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.
KW  - Bacillaceae
KW  - Biotechnological application
KW  - Broad pH spectrum
KW  - Subtilases
KW  - Subtilisin
Y1  - 2023
U6  - http://dx.doi.org/10.1002/2211-5463.13701
SN  - 2211-5463
N1  - Corresponding author: Petra Siegert
VL  - 13
IS  - 11
SP  - 2035
EP  - 2046
PB  - Wiley
CY  - Hoboken, NJ
ER  - 
TY  - JOUR
A1  - Falkenberg, Fabian
A1  - Bott, Michael
A1  - Bongaerts, Johannes
A1  - Siegert, Petra
T1  - Phylogenetic survey of the subtilase family and a data-mining-based search for new subtilisins from Bacillaceae
JF  - Frontiers in Microbiology
N2  - The subtilase family (S8), a member of the clan SB of serine proteases are ubiquitous in all kingdoms of life and fulfil different physiological functions. Subtilases are divided in several groups and especially subtilisins are of interest as they are used in various industrial sectors. Therefore, we searched for new subtilisin sequences of the family Bacillaceae using a data mining approach. The obtained 1,400 sequences were phylogenetically classified in the context of the subtilase family. This required an updated comprehensive overview of the different groups within this family. To fill this gap, we conducted a phylogenetic survey of the S8 family with characterised holotypes derived from the MEROPS database. The analysis revealed the presence of eight previously uncharacterised groups and 13 subgroups within the S8 family. The sequences that emerged from the data mining with the set filter parameters were mainly assigned to the subtilisin subgroups of true subtilisins, high-alkaline subtilisins, and phylogenetically intermediate subtilisins and represent an excellent source for new subtilisin candidates.
Y1  - 2022
U6  - http://dx.doi.org/10.3389/fmicb.2022.1017978
SN  - 1664-302X
VL  - 2022
IS  - 13
PB  - Frontiers
CY  - Lausanne
ER  - 
TY  - JOUR
A1  - Dünnwald, Thomas
A1  - Demir, Ayhan S.
A1  - Siegert, Petra
A1  - Pohl, Martina
A1  - Müller, Michael
T1  - ChemInform Abstract: Enantioselective synthesis of (S)-2-Hydroxypropanone derivatives by Benzoylformate Decarboxylase Catalyzed C—C Bond Formation
JF  - Cheminform
Y1  - 2001
SN  - 1522-2667 (E-Journal); 0931-7597 (Print)
VL  - Vol. 32
IS  - Iss. 4
SP  - Publ. online
ER  - 
TY  - JOUR
A1  - Dünnwald, Thomas
A1  - Demir, Ayhan S.
A1  - Siegert, Petra
A1  - Pohl, Martina
A1  - Müller, Michael
T1  - Enantioselective Synthesis of (S)-2-Hydroxypropanone Derivatives by Benzoylformate Decarboxylase Catalyzed C−C Bond Formation
JF  - European journal of organic chemistry
Y1  - 2000
SN  - 0365-5490 (E-Journal); 1099-0690 (E-Journal); 0075-4617 (Print); 0170-2041 (Print); 0947-3440 (Print); 1434-193X (Print); 1434-243X (Print)
VL  - Vol. 2000
IS  - Iss. 11
SP  - 2161
EP  - 2170
ER  - 
TY  - JOUR
A1  - Dünkelmann, Pascal
A1  - Kolter-Jung, Doris
A1  - Nitsche, Adam
A1  - Demir, Ayhan S.
A1  - Siegert, Petra
A1  - Lingen, Bettina
A1  - Baumann, Martin
A1  - Pohl, Martina
A1  - Müller, Michael
T1  - Development of a donor-acceptor concept for enzymatic cross-coupling reactions of adehydes : the first asymmetric cross-benzoin condensation
JF  - Journal of the American Chemical Society
Y1  - 2002
SN  - 1520-5126 (E-Journal); 0002-7863 (Print)
VL  - Vol. 124
SP  - 12084
EP  - 12085
ER  - 
TY  - PAT
A1  - Bessler, Cornelius
A1  - Maurer, Karl-Heinz
A1  - Merkel, Marion
A1  - Siegert, Petra
A1  - Wieland, Susanne
T1  - Subtilisin from Bacillus Pumilus and detergent and cleaning agents containing said novel subtilisin [US Patentanmeldung / Internationale Patentanmeldung]
Y1  - 2009
SP  - 1
EP  - 39
PB  - USPTO; WIPO
CY  - Washington; Genf
ER  -