TY - JOUR

T1 - Biocatalyst Immobilization by Anchor Peptides on an Additively Manufacturable Material

AU - Büscher, Niclas

AU - Sayoga, Giovanni V.

AU - Rübsam, Kristin

AU - Jakob, Felix

AU - Schwaneberg, Ulrich

AU - Kara, Selin

AU - Liese, Andreas

N1 - Funding Information: This research was made possible by grant of Behörde für Wissenschaft, Forschung und Gleichstellung in Hamburg. We acknowledge Marcel Fassbender from the University of Hamburg, Department of Technical and Macromolecular Chemistry, for his support with additive manufacturing of PET. We thank Dipl.-Ing. Victor Baudron from Hamburg University of Technology, Institute of Thermal Separation Processes, for Brunauer–Emmett–Teller (BET) measurements of PET powder and Victoria Büschler, Alexander Mook, and Maren Breuer for their support in lab work.

PY - 2019/9/20

Y1 - 2019/9/20

N2 - Additive manufacturing refers to manufacturing methods, which are being used to build up three-dimensional (3D) structures by adding a certain material stepwise onto a support. Nowadays, these manufacturing methods can be more material- and cost-efficient compared to conventional methods and allowing the defined production of a wide variety of 3D structures using computer aided design (CAD). A broad range of materials can be additively manufactured (AM) resulting in specific properties and highly diverse structures making them promising matrices for the utilization as enzyme carriers. The variety of materials offers the possibility to select materials with properties needed for particular biocatalytic processes. This is especially true for hybrid reactor concepts where multiple operations, including catalytic reactions and downstream processes, are combined into a single apparatus. For the enzymatic decarboxylation of ferulic acid, polyethylene terephthalate (PET) has been chosen as an additively manufacturable carrier material for the immobilization of phenolic acid decarboxylase (PAD) from Mycobacterium columbiense. The genetic fusion of PAD with anchor peptides enabled the adsorptive immobilization on PET. Starting from an immobilizate activity of 0.39 ± 0.19 U m-2 and a conversion of 19.2 ± 3.7% after 2 h the optimization of the peptide and spacer sequence between anchor peptide and PAD resulted in immobilizates with activities up to 1.80 ± 0.41 U m-2 and conversions of 59.9 ± 3.9% after 2 h. Moreover, within this study integrating an in situ product removal, enabled by an extraction with n-heptane, the altering of surface hydrophobicity of PET and a conversion of 88.0 ± 3.8% after 2 h could be observed.

AB - Additive manufacturing refers to manufacturing methods, which are being used to build up three-dimensional (3D) structures by adding a certain material stepwise onto a support. Nowadays, these manufacturing methods can be more material- and cost-efficient compared to conventional methods and allowing the defined production of a wide variety of 3D structures using computer aided design (CAD). A broad range of materials can be additively manufactured (AM) resulting in specific properties and highly diverse structures making them promising matrices for the utilization as enzyme carriers. The variety of materials offers the possibility to select materials with properties needed for particular biocatalytic processes. This is especially true for hybrid reactor concepts where multiple operations, including catalytic reactions and downstream processes, are combined into a single apparatus. For the enzymatic decarboxylation of ferulic acid, polyethylene terephthalate (PET) has been chosen as an additively manufacturable carrier material for the immobilization of phenolic acid decarboxylase (PAD) from Mycobacterium columbiense. The genetic fusion of PAD with anchor peptides enabled the adsorptive immobilization on PET. Starting from an immobilizate activity of 0.39 ± 0.19 U m-2 and a conversion of 19.2 ± 3.7% after 2 h the optimization of the peptide and spacer sequence between anchor peptide and PAD resulted in immobilizates with activities up to 1.80 ± 0.41 U m-2 and conversions of 59.9 ± 3.9% after 2 h. Moreover, within this study integrating an in situ product removal, enabled by an extraction with n-heptane, the altering of surface hydrophobicity of PET and a conversion of 88.0 ± 3.8% after 2 h could be observed.

KW - additive manufacturing

KW - adsorptive immobilization

KW - anchor peptides

KW - biocatalysis

KW - protein engineering

UR - http://www.scopus.com/inward/record.url?scp=85070584835&partnerID=8YFLogxK

U2 - 10.1021/acs.oprd.9b00152

DO - 10.1021/acs.oprd.9b00152

M3 - Article

AN - SCOPUS:85070584835

VL - 23

SP - 1852

EP - 1859

JO - Organic Process Research and Development

JF - Organic Process Research and Development

SN - 1083-6160

IS - 9

ER -