TY - JOUR

T1 - Immobilization and characterization of benzoylformate decarboxylase from Pseudomonas putida on spherical silica carrier

AU - Peper, Stephanie

AU - Kara, Selin

AU - Long, Wei Sing

AU - Liese, Andreas

AU - Niemeyer, Bernd

N1 - Funding Information: Acknowledgments We thank the Deutsch Forschungsgemeinschaft DFG for the financial support of the project. Prof. Dr. Martina Pohl (Institute of Molecular Enzyme Technology, Heinrich-Heine University of Düsseldorf, Research Center Jülich) is gratefully acknowledged for kindly providing the variant BFD A460I-F464I.

PY - 2011/8

Y1 - 2011/8

N2 - If an adequate biocatalyst is identified for a specific reaction, immobilization is one possibility to further improve its properties. The immobilization allows easy recycling, improves the enzyme performance, and it often enhances the stability of the enzyme. In this work, the immobilization of the benzoylformate decarboxylase (BFD) variant, BFD A460I-F464I, from Pseudomonas putida was accomplished on spherical silica. Silicagel is characterized by its high mechanical stability, which allows its application in different reactor types without restrictions. The covalently bound enzyme was characterized in terms of its activity, stability, and kinetics for the formation of chiral 2-hydroxypropiophenone (2-HPP) from benzaldehyde and acetaldehyde. Moreover, temperature as well as pressure dependency of immobilized BFD A460I-F464I activity and enantioselectivity were analyzed. The used wide-pore silicagel shows a good accessibility of the immobilized enzyme. The activity of the immobilized BFD A460I-F464I variant was determined to be 70% related to the activity of the free enzyme. Thereby, the enantioselectivity of the enzyme was not influenced by the immobilization. In addition, a pressure-induced change in stereoselectivity was found both for the free and for the immobilized enzyme. With increasing pressure, the enantiomeric excess (ee) of (R)-2-HPP can be increased from 44% (0.1 MPa) to 76% (200 MPa) for the free enzyme and from 43% (0.1 MPa) to 66% (200 MPa) for the immobilized enzyme.

AB - If an adequate biocatalyst is identified for a specific reaction, immobilization is one possibility to further improve its properties. The immobilization allows easy recycling, improves the enzyme performance, and it often enhances the stability of the enzyme. In this work, the immobilization of the benzoylformate decarboxylase (BFD) variant, BFD A460I-F464I, from Pseudomonas putida was accomplished on spherical silica. Silicagel is characterized by its high mechanical stability, which allows its application in different reactor types without restrictions. The covalently bound enzyme was characterized in terms of its activity, stability, and kinetics for the formation of chiral 2-hydroxypropiophenone (2-HPP) from benzaldehyde and acetaldehyde. Moreover, temperature as well as pressure dependency of immobilized BFD A460I-F464I activity and enantioselectivity were analyzed. The used wide-pore silicagel shows a good accessibility of the immobilized enzyme. The activity of the immobilized BFD A460I-F464I variant was determined to be 70% related to the activity of the free enzyme. Thereby, the enantioselectivity of the enzyme was not influenced by the immobilization. In addition, a pressure-induced change in stereoselectivity was found both for the free and for the immobilized enzyme. With increasing pressure, the enantiomeric excess (ee) of (R)-2-HPP can be increased from 44% (0.1 MPa) to 76% (200 MPa) for the free enzyme and from 43% (0.1 MPa) to 66% (200 MPa) for the immobilized enzyme.

KW - Benzoylformate decarboxylase

KW - Enantioselectivity

KW - Immobilization

KW - Silicagel

UR - http://www.scopus.com/inward/record.url?scp=79960452552&partnerID=8YFLogxK

U2 - 10.1007/s00449-011-0516-0

DO - 10.1007/s00449-011-0516-0

M3 - Article

C2 - 21286757

AN - SCOPUS:79960452552

VL - 34

SP - 671

EP - 680

JO - Bioprocess and Biosystems Engineering

JF - Bioprocess and Biosystems Engineering

SN - 1615-7591

IS - 6

ER -