TY  - PAT
A1  - Siegert, Petra
A1  - Merkel, Marion
A1  - Hellmuth, Hendrik
A1  - O'Connell, Timothy
A1  - Maurer, Karl-Heinz
T1  - Stabilisierte flüssige enzymhaltige Tensidzubereitung (Einsatz einer das hydrolytische Enzym stabilisierende Komponente, die ein Monosaccharidglycerat umfasst) [Offenlegungsschrift]
T1  - Stabilized liquid enzyme-containing surfactant preparation (using a component which comprises a monosaccharide glycerate and which stabilizes the hydrolytic enzyme) [Europäische Patentanmeldung / Internationale Patentanmeldung]
Y1  - 2012
SP  - 1
EP  - 17
PB  - Deutsches Patent- und Markenamt / Europäisches Patentamt / WIPO
CY  - München / Den Hague / Genf
ER  - 
TY  - PAT
A1  - Siegert, Petra
A1  - Merkel, Marion
A1  - Hellmuth, Hendrik
A1  - O'Connell, Timothy
A1  - Maurer, Karl-Heinz
T1  - Stabilisierte flüssige enzymhaltige Tensidzubereitung (durch den Einsatz einer das hydrolytische Enzym stabilisierende Komponente, die eine mehrfach substituierte Benzolcarbonsäure umfasst, die an mindestens zwei Kohlenstoffatomen des Benzolrestes eine Carboxylgruppe aufweist) [Offenlegungsschrift]
T1  - Stabilized liquid tenside preparation comprising enzymes (a component comprising a multiply substituted benzyl carboxylic acid comprising a carboxyl group on at least two carbon atoms of the benzyl radical is used for stabilizing the hydrolytic enzyme) [Europäische Patentanmeldung / Internationale Patentanmeldung]
Y1  - 2012
SP  - 1
EP  - 16
PB  - Deutsches Patent- und Markenamt / Europäisches Patentamt / WIPO
CY  - München / Den Hague / Genf
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  - Haeger, Gerrit
A1  - Jolmes, Tristan
A1  - Oyen, Sven
A1  - Jaeger, Karl-Erich
A1  - Bongaerts, Johannes
A1  - Schörken, Ulrich
A1  - Siegert, Petra
T1  - Novel recombinant aminoacylase from Paraburkholderia monticola capable of N-acyl-amino acid synthesis
JF  - Applied Microbiology and Biotechnology
N2  - N-Acyl-amino acids can act as mild biobased surfactants, which are used, e.g., in baby shampoos. However, their chemical synthesis needs acyl chlorides and does not meet sustainability criteria. Thus, the identification of biocatalysts to develop greener synthesis routes is desirable. We describe a novel aminoacylase from Paraburkholderia monticola DSM 100849 (PmAcy) which was identified, cloned, and evaluated for its N-acyl-amino acid synthesis potential. Soluble protein was obtained by expression in lactose autoinduction medium and co-expression of molecular chaperones GroEL/S. Strep-tag affinity purification enriched the enzyme 16-fold and yielded 15 mg pure enzyme from 100 mL of culture. Biochemical characterization revealed that PmAcy possesses beneficial traits for industrial application like high temperature and pH-stability. A heat activation of PmAcy was observed upon incubation at temperatures up to 80 °C. Hydrolytic activity of PmAcy was detected with several N-acyl-amino acids as substrates and exhibited the highest conversion rate of 773 U/mg with N-lauroyl-L-alanine at 75 °C. The enzyme preferred long-chain acyl-amino-acids and displayed hardly any activity with acetyl-amino acids. PmAcy was also capable of N-acyl-amino acid synthesis with good conversion rates. The best synthesis results were obtained with the cationic L-amino acids L-arginine and L-lysine as well as with L-leucine and L-phenylalanine. Exemplarily, L-phenylalanine was acylated with fatty acids of chain lengths from C8 to C18 with conversion rates of up to 75%. N-lauroyl-L-phenylalanine was purified by precipitation, and the structure of the reaction product was verified by LC–MS and NMR.
KW  - Chaperone
KW  - Biocatalysis
KW  - Aminoacylase
KW  - Acylation
KW  - Acyl-amino acids
KW  - Biosurfactants
Y1  - 2024
U6  - http://dx.doi.org/10.1007/s00253-023-12868-8
SN  - 1432-0614
N1  - Corresponding author: Petra Siegert
IS  - 108
PB  - Springer
CY  - Berlin
ER  - 
TY  - JOUR
A1  - Haeger, Gerrit
A1  - Probst, Johanna
A1  - Jaeger, Karl-Erich
A1  - Bongaerts, Johannes
A1  - Siegert, Petra
T1  - Novel aminoacylases from Streptomyces griseus DSM 40236 and their recombinant production in Streptomyces lividans
JF  - FEBS Open Bio
N2  - Amino acid-based surfactants are valuable compounds for cosmetic formulations. The chemical synthesis of acyl-amino acids is conventionally performed by the Schotten-Baumann reaction using fatty acyl chlorides, but aminoacylases have also been investigated for use in biocatalytic synthesis with free fatty acids. Aminoacylases and their properties are diverse; they belong to different peptidase families and show differences in substrate specificity and biocatalytic potential. Bacterial aminoacylases capable of synthesis have been isolated from Burkholderia, Mycolicibacterium, and Streptomyces. Although several proteases and peptidases from S. griseus have been described, no aminoacylases from this species have been identified yet. In this study, we investigated two novel enzymes produced by S. griseus DSM 40236ᵀ . We identified and cloned the respective genes and recombinantly expressed an α-aminoacylase (EC 3.5.1.14), designated SgAA, and an ε-lysine acylase (EC 3.5.1.17), designated SgELA, in S. lividans TK23. The purified aminoacylase SgAA was biochemically characterized, focusing on its hydrolytic activity to determine temperature- and pH optima and stabilities. The aminoacylase could hydrolyze various acetyl-amino acids at the Nα -position with a broad specificity regarding the sidechain. Substrates with longer acyl chains, like lauroyl-amino acids, were hydrolyzed to a lesser extent. Purified aminoacylase SgELA specific for the hydrolysis of Nε -acetyl-L-lysine was unstable and lost its enzymatic activity upon storage for a longer period but could initially be characterized. The pH optimum of SgELA was pH 8.0. While synthesis of acyl-amino acids was not observed with SgELA, SgAA catalyzed the synthesis of lauroyl-methionine.
KW  - Streptomyces lividans
KW  - recombinant expression
KW  - Streptomyces griseus
KW  - ε-lysine acylase
KW  - α-aminoacylase
Y1  - 2023
U6  - http://dx.doi.org/10.1002/2211-5463.13723
SN  - 2211-5463
N1  - Corresponding author: Petra Siegert
VL  - 13
IS  - 12
SP  - 2224
EP  - 2238
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  - PAT
A1  - Siegert, Petra
A1  - Mussmann, Nina
A1  - O'Connell, Timothy
A1  - Maurer, Karl-Heinz
T1  - Neue Proteasen und Mittel enthaltend diese Proteasen [Offenlegungsschrift]
T1  - Novel proteases and means containing said proteases [Internationale Patentanmeldung]
Y1  - 2010
SP  - 1
EP  - 30
PB  - Deutsches Patent- und Markenamt / WIPO
CY  - München / Genf
ER  - 
TY  - PAT
A1  - Siegert, Petra
A1  - Baumstark, Rebecca
A1  - Kluin, Cornelia
A1  - O'Connell, Timothy
A1  - Maurer, Karl-Heinz
A1  - Hellmuth, Hendrik
T1  - Neue Proteasen und Mittel enthaltend diese Proteasen [Offenlegungsschrift]
Y1  - 2010
SP  - 1
EP  - 30
PB  - Deutsches Patent- und Markenamt
CY  - München
ER  - 
TY  - PAT
A1  - Siegert, Petra
A1  - Spitz, Astrid
A1  - Maurer, Karl-Heinz
T1  - Neue Proteasen und Mittel enthaltend diese Proteasen [Offenlegungsschrift]
T1  - Novel proteases and compositions comprising these proteases [Internationale Patentanmeldung]
Y1  - 2010
SP  - 1
EP  - 31
PB  - Deutsches Patentamt / WIPO
CY  - München / Genf
ER  - 
TY  - PAT
A1  - Siegert, Petra
A1  - Merkel, Marion
A1  - Kluin, Cornelia
A1  - Maurer, Karl-Heinz
A1  - O'Connell, Timothy
A1  - Wieland, Susanne
A1  - Hellmuth, Hendrik
T1  - Neue Proteasen und diese enthaltende Mittel [Offenlegungsschrift]
T1  - Novel proteases and compositions comprising them [Europäische Patentanmeldung / Internationale Patentanmeldung]
Y1  - 2011
SP  - 1
EP  - 21
PB  - Deutsches Patent- und Markenamt / Europäisches Patentamt / WIPO
CY  - München / Den Hague / Genf
ER  - 
TY  - PAT
A1  - Siegert, Petra
A1  - Wieland, Susanne
A1  - Engelskirchen, Julia
A1  - Merkel, Marion
A1  - Maurer, Karl-Heinz
A1  - Bessler, Cornelius
T1  - Neue Alkalische Protease aus Bacillus gibsonii und Wasch- und Reinigungsmittel enthaltend diese neue Alkalische Protease [Offenlegungsschrift]
T1  - Novel alkaline protease from Bacillus Gibsonii and washing and cleaning agents containing said novel alkaline protease [Europäische Patentanmeldung / US Patentanmeldung / Internationale Patentanmeldung]
Y1  - 2008
SP  - 1
EP  - 51
PB  - Deutsches Patentamt / Europäisches Patentamt / WIPO
CY  - München / Den Hague / Genf
ER  - 
TY  - PAT
A1  - Merkel, Marion
A1  - Weber, Angrit
A1  - Siegert, Petra
A1  - Wieland, Susanne
A1  - Maurer, Karl-Heinz
A1  - Bessler, Cornelius
T1  - Neue Alkalische Protease aus Bacillus gibsonii und Wasch- und Reinigungsmittel enthaltend diese neue Alkalische Protease [Offenlegungsschrift]
T1  - Novel alkaline protease from Bacillus Gibsonii and washing and cleaning agents containing said novel alkaline protease [Internationale Patentanmeldung]
Y1  - 2007
SP  - 1
EP  - 46
PB  - Deutsches Patent- und Markenamt / WIPO
CY  - München / Genf
ER  - 
TY  - PAT
A1  - Berg, Gabriele
A1  - Bessler, Cornelius
A1  - Gerlach, Jochen
A1  - Gübitz, Georg
A1  - Heumann, Sonja
A1  - Karl, Wolfgang
A1  - Maurer, Karl-Heinz
A1  - Remler, Peter
A1  - Ribitsch, Doris
A1  - Schwab, Helmut
A1  - Siegert, Petra
A1  - Wieland, Susanne
T1  - Mittel enthaltend Proteasen aus Stenotrophomonas maltophilia [Offenlegungsschrift]
T1  - Agents containing proteases from Stenotrophomonas maltophilia [Europäische Patentanmeldung / US Patentanmeldung / Internationale Patentanmeldung]
Y1  - 2009
SP  - 1
EP  - 47
PB  - Deutsches Patent- und Markenamt / Europäisches Patentamt / USPTO / WIPO
CY  - München / Den Hague / Washington / Genf
ER  - 
TY  - PAT
A1  - O'Connell, Timothy
A1  - Siegert, Petra
A1  - Maurer, Karl-Heinz
A1  - Schiedel, Marc-Steffen
A1  - Vockenroth, Inga Kerstin
T1  - Method for improving the cleaning action of a detergent or cleaning agent [Internationale Patentanmeldung]
T1  - Verfahren zur Verbesserung der Reinigungsleistung eines Wasch- oder Reinigungsmittels
Y1  - 2010
SP  - 1
EP  - 15
PB  - WIPO
CY  - Genf
ER  - 
TY  - PAT
A1  - Banowski, Bernhard
A1  - Wadle, Armin
A1  - Siegert, Petra
A1  - Sättler, Andrea
T1  - Lipase-Inhibitoren in Deodorantien und Antitranspirantien [Offenlegungsschrift]
T1  - Lipase inhibitors in deodorants and antiperspirants [Europäische Patentanmeldung]
Y1  - 2004
SP  - 1
EP  - 22
PB  - Deutsches Patent- und Markenamt / Europäisches Patentamt
CY  - München / Den Hague
ER  - 
TY  - PAT
A1  - Siegert, Petra
A1  - Schwaneberg, Ulrich
A1  - Martinez Moya, Ronny
A1  - Merkel, Marion
A1  - Spitz, Astrid
A1  - Wieland, Susanne
A1  - Hellmuth, Hendrik
A1  - Maurer, Karl-Heinz
T1  - Leistungsverbesserte Proteasevariante [Offenlegungsschrift]
T1  - Performance-enhanced protease variant [Europäische Patentanmeldung / Internationale Patentanmeldung]
Y1  - 2012
SP  - 1
EP  - 29
PB  - Deutsches Patent- und Markenamt / Europäisches Patentamt / WIPO
CY  - München / Den Hague / Genf
ER  - 
TY  - PAT
A1  - Bessler, Cornelius
A1  - Evers, Stefan
A1  - Maurer, Karl-Heinz
A1  - Merkel, Marion
A1  - Siegert, Petra
A1  - Weber, Angrit
A1  - Wieland, Susanne
T1  - Leistungsverbesserte Proteasen und Wasch- und Reinigungsmittel enthaltend diese Proteasen [Offenlegungsschrift]
T1  - Improved-performance proteases and detergents and cleaning agents comprising said proteases [Internationale Patentanmeldung]
Y1  - 2009
SP  - 1
EP  - 41
PB  - Deutsches Patent- und Markenamt / WIPO
CY  - München / Genf
ER  - 
TY  - PAT
A1  - Wieland, Susanne
A1  - Siegert, Petra
A1  - Spitz, Astrid
A1  - Maurer, Karl-Heinz
A1  - O'Connell, Timothy
A1  - Prüser, Inken
A1  - Schiedel, Marc-Steffen
A1  - Eiting, Thomas
A1  - Sendor-Müller, Dorota
A1  - Bastigkeit, Thorsten
A1  - Benda, Konstantin
A1  - Müller, Sven
T1  - Lagerstabiles flüssiges Wasch- oder Reinigungsmittel enthaltend Proteasen [Offenlegungsschrift]
T1  - Storage-stable liquid washing or cleaning agent containing proteases [Europäische Patentanmeldung / Internationale Patentanmeldung]
Y1  - 2011
SP  - 1
EP  - 25
PB  - Deutsches Patent- und Markenamt / Europäisches Patentamt / WIPO
CY  - München / Den Hague / Genf
ER  - 
TY  - JOUR
A1  - Martinez, Ronny
A1  - Jakob, Felix
A1  - Tu, Ran
A1  - Siegert, Petra
A1  - Maurer, Karl-Heinz
A1  - Schwaneberg, Ulrich
T1  - Increasing activity and thermal resistance of Bacillus gibsonii alkaline protease (BgAP) by directed evolution
JF  - Biotechnology and bioengineering
Y1  - 2013
SN  - 1097-0290 (E-Journal); 0006-3592 (Print); 0368-1467 (Print)
VL  - Vol. 110
IS  - Iss. 3
SP  - 711
EP  - 720
PB  - Wiley
CY  - Weinheim
ER  - 
TY  - JOUR
A1  - Ribitsch, D.
A1  - Heumann, S.
A1  - Trotscha, E.
A1  - Herrero Acero, E.
A1  - Greimel, K.
A1  - Leber, R.
A1  - Birger-Gruenberger, R.
A1  - Deller, S.
A1  - Eiteljoerg, I.
A1  - Remler, P.
A1  - Weber, Th.
A1  - Siegert, Petra
A1  - Maurer, Karl-Heinz
A1  - Donelli, I.
A1  - Freddi, G.
A1  - Schwab, H.
A1  - Guebitz, G. M.
T1  - Hydrolysis of polyethyleneterephthalate by p-nitrobenzylesterase from Bacillus subtilis
JF  - Biotechnology progress
Y1  - 2011
SN  - 1520-6033 (E-Journal); 8756-7938 (Print)
VL  - Vol. 27
IS  - Iss. 4
SP  - 951
EP  - 960
PB  - Wiley
CY  - Hoboken
ER  -