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Publications (2) View all
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Article: Discovery and Protein Engineering of Biocatalysts for Organic Synthesis
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ABSTRACT: Modern tools for enzyme discovery and protein engineering substantially broadened the number of enzymes applicable for biocatalysis and helped to alter their properties such as substrate range, enantioselectivity, and stability under process conditions. In addition, these methods also enabled one to explore reactions for organic synthesis for which no suitable enzymes were available until recently. This review provides a summary of the different concepts and technologies, which are exemplified for various enzymes.Advanced Synthesis & Catalysis 09/2011; 353(13):2191 - 2215. · 6.05 Impact Factor -
Article: Pseudomonas putida esterase contains a GGG(A)X-motif confering activity for the kinetic resolution of tertiary alcohols.
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ABSTRACT: An esterase from Pseudomonas putida JD1 (PPE) was successfully cloned, actively expressed in Escherichia coli, and characterized. It was discovered that PPE is more active towards short-chain esters, hydrolyzed δ-valerolactone, and ε-caprolactone and was most active at 37°C and pH 8. After purification to homogeneity by Ni-NTA-assisted affinity chromatography, the kinetic parameters K(M) and k(cat) were determined for p-nitrophenyl acetate and butyrate, respectively, showing better catalytic efficiency for hydrolysis of the acetate residue. Investigation of the protein sequence revealed not only the classical catalytic triad for carboxylesterases, additionally the interesting GGG(A)X-motif, which is associated to activity towards tertiary alcohols, was found. Indeed, enzymatic activity was shown for a set of different tertiary alcohols with enantioselectivities up to E = 20, suggesting PPE to be a promising biocatalyst. In addition, PPE also hydrolyzed 4-hydroxyphenyl acetate, the product of a Baeyer-Villiger monooxygenase-catalyzed oxidation of 4-hydroxyacetophenone with a specific activity of 34.36 U/mg suggesting a physiological role in P. putida JD1.Applied Microbiology and Biotechnology 07/2011; 93(3):1119-26. · 3.42 Impact Factor
