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  - O'Connell, Timothy
A1  - Siegert, Petra
A1  - Evers, Stefan
A1  - Bongaerts, Johannes
A1  - Weber, Thomas
A1  - Maurer, Karl-Heinz
A1  - Bessler, Cornelius
T1  - Wasch- oder Reinigungsmittel mit gesteigerter Waschkraft [Offenlegungsschrift]
T1  - Method from improving the cleaning action of a detergent of cleaning agent [US Patentanmeldung]
Y1  - 2010
SP  - 1
EP  - 34
PB  - Deutsches Patentamt
CY  - München
ER  - 
TY  - PAT
A1  - Siegert, Petra
A1  - Spitz, Astrid
A1  - Maurer, Karl-Heinz
T1  - Wasch- und Reinigungsmittel enthaltend Proteasen aus Bacillus pumilus [Offenlegungsschrift]
T1  - Detergents and cleaning agents containing proteases from bacillus pumilus [Europäische Patentschrift / Internationale Patentanmeldung]
Y1  - 2010
SP  - 1
EP  - 20
PB  - Deutsches Patentamt / Europäisches Patentamt / WIPO
CY  - München / Den Hague / Genf
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  - 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  - 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  - JOUR
A1  - Ribitsch, D.
A1  - Karl, W.
A1  - Birner-Gruenberger, R.
A1  - Gruber, K.
A1  - Eiteljoerg, I.
A1  - Remler, P.
A1  - Wieland, S.
A1  - Siegert, Petra
A1  - Maurer, Karl-Heinz
A1  - Schwab, H.
T1  - C-terminal truncation of a metagenome-derived detergent protease for effective expression in E. coli
JF  - Journal of biotechnology
N2  - Recently, a new alkaline protease named HP70 showing highest homology to extracellular serine proteases of Stenotrophomonas maltophilia and Xanthomonas campestris was found in the course of a metagenome screening for detergent proteases (Niehaus et al., submitted for publication). Attempts to efficiently express the enzyme in common expression hosts had failed. This study reports on the realization of overexpression in Escherichia coli after structural modification of HP70. Modelling of HP70 resulted in a two-domain structure, comprising the catalytic domain and a C-terminal domain which includes about 100 amino acids. On the basis of the modelled structure the enzyme was truncated by deletion of most of the C-terminal domain yielding HP70-C477.

This structural modification allowed effective expression of active enzyme using E. coli BL21-Gold as the host. Specific activity of HP70-C477 determined with suc-l-Ala-l-Ala-l-Pro-l-Phe-p-nitroanilide as the substrate was 30 ± 5 U/mg compared to 8 ± 1 U/mg of the native enzyme. HP70-C477 was most active at 40 °C and pH 7–11; these conditions are prerequisite for a potential application as detergent enzyme. Determination of kinetic parameters at 40 °C and pH = 9.5 resulted in KM = 0.23 ± 0.01 mM and kcat = 167.5 ± 3.6 s⁻¹. MS-analysis of peptide fragments obtained from incubation of HP70 and HP70-C477 with insulin B indicated that the C-terminal domain influences the cleavage preferences of the enzyme. Washing experiments confirmed the high potential of HP70-C477 as detergent protease.
Y1  - 2010
U6  - http://dx.doi.org/10.1016/j.jbiotec.2010.09.947
SN  - 1873-4863 (E-Journal); 0168-1656 (Print)
VL  - 150
IS  - 3
SP  - 408
EP  - 416
PB  - Elsevier
CY  - Amsterdam
ER  -