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Technical University of Munich
Technical University of Munich

Surface functionalisation of nano-scale membrane reactors for multienzyme syntheses

Ludwig Klermund, doctoral thesis Technische Universität München, 2017

By entrapping enzymes in polymeric vesicles with a selectively permeable membrane, incompatible reactions of multienzyme syntheses can be spatially separated. A new method for immobilizing enzymes on the vesicle surface was developed making the surrounding environment available as additional reaction space. Using hydrophobic peptide anchors up to 2320 proteins were immobilized per vesicle. Immobilization and selective mass transport improved the yield of an exemplary reaction cascade 2.2-fold.

Publications

  • Schwarzer TS, Klermund L, Wang G, Castiglione K (2018): Membrane functionalization of polymersomes: allevating mass transport limitations by integrating multiple selective membrane transporters for the diffusion of chemically diverse molecules. Nanotechnol 29: 44LT01.
  • Klermund L, Castiglione K (2018): Polymersomes as nanoreactors for preparative biocatalytic applications: current challenges and future perspectives. Bioproc Biosys Eng 41:1233–1246.
  • Poschenrieder ST, Klermund L, Langer B, Castiglione K (2017): Determination of Permeability Coefficients of Polymersomal Membranes for Hydrophilic Molecules. Langmuir, 33: 6011-6020.
  • Klermund L, Poschenrieder TS, Castiglione K (2017): Biocatalysis in Polymersomes: Improving Multienzyme Cascades with Incompatible Reaction Steps by Compartmentalization. ACS Catal 7: 3900-3904.
  • Schmideder A, Schottroff F, Klermund L, Castiglione K, Weuster-Botz D (2017): Studies on the enzymatic synthesis of N-acetylneuraminic acid with continuously operated enzyme membrane reactors on a milliliter scale. Biochem Eng J 119: 9 - 19.
  • Klermund L, Poschenrieder ST, Castiglione K (2016): Simple surface functionalization of polymersomes using non-antibacterial peptide anchors. J Nanobiotechnol 14:48.
  • Klermund L, Riederer A, Hunger A, Castiglione K (2016): Protein engineering of a bacterial N-acyl-D-glucosamine 2-epimerase for improved stability under process conditions. Enzyme Microb Technol 87-88: 70-78.
  • Klermund L, Riederer A, Groher A, Castiglione K (2015): High-level soluble expression of a bacterial N-acyl-D-glucosamine 2-epimerase in recombinant Escherichia coli. Protein Expr Purif 111: 36-41.
  • Klermund L, Groher A, Castiglione K (2013): New N-acyl-D-glucosamine 2-epimerases from cyanobacteria with high activity in the absence of ATP and low inhibition by pyruvate. J Biotechnol 168: 256-263.
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