Endosporen-basierte Immobilisierung von Lipasen für die anspruchsvolle Biokatalyse
Zusammenfassung der Projektergebnisse
Due to their resting and resistant properties, B. subtilis SporoBeads have great potential as a protein autoimmobilisation platform. They are of interest in many areas of research, including medical, chemical and food applications. In this project, we have applied a number of modification strategies to enhance the potential and improve the outcome. We have shown that the PcotYZ promoter is still unrivalled for gene expression. We also found novel anchor proteins in the deepest protein layer, the inner coat. Replicative plasmids were used for the first time in this system and were able to increase reporter activity 10-fold. Our discovery of a functional crust opens the door to any kind of biotechnological application and actual implementation of this system, as it is not a GMO. During the course of this project, we also demonstrated that lipases, contrary to reports in the literature, are not capable of silyl ether conversion due to the nature of their active site and are therefore not suitable biocatalysts for silyl ether conversion. However, we have identified two novel enzymes, SilE-R and SilE-S, which are able to hydrolyse silyl ethers in an enantiospecific manner. Thus, for the first time, biocatalysts for the enantiospecific hydrolysis of silyl ethers have been identified and characterised. We were able to show which molecular determinants are necessary for the successful enzymatic conversion of silyl ethers, which will facilitate the identification of further biocatalysts for the conversion of silyl ethers in the future. The discovery of these enzymes and the insights gained from this project are of enormous importance for the development of enzyme-based enantiospecific protective group strategies (and other applications) using silyl ethers. Finally, we demonstrated that the SporoBeads system is suitable for immobilising SilE-R. The resulting SilE-R-SporoBeads showed competitive activity with conventionally immobilised enzymes, while being superior in terms of handling and reusability.
Projektbezogene Publikationen (Auswahl)
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Lipase‐Mediated Conversion of Protecting Group Silyl Ethers: An Unspecific Side Reaction. ChemBioChem, 24(18).
Pick, Lisa M.; Wenzlaff, Jessica; Yousefi, Mohammad; Davari, Mehdi D. & Ansorge‐Schumacher, Marion B.
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SilE‐R and SilE‐S—DABB Proteins Catalysing Enantiospecific Hydrolysis of Organosilyl Ethers. Angewandte Chemie International Edition, 63(25).
Pick, Lisa M.; Oehme, Viviane; Hartmann, Julia; Wenzlaff, Jessica; Tang, Qingyun; Grogan, Gideon & Ansorge‐Schumacher, Marion B.
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SilE−R und SilE−S – DABB‐Proteine mit der Fähigkeit zur enantiospezifischen Hydrolyse von Organosilylethern. Angewandte Chemie, 136(25).
Pick, Lisa M.; Oehme, Viviane; Hartmann, Julia; Wenzlaff, Jessica; Tang, Qingyun; Grogan, Gideon & Ansorge‐Schumacher, Marion B.
