ESR 6 - Smart enzyme immobilization for biocatalytic microreactor development

Host: TUG; Supervisor: Bernd Nidetzky

The main objectives of the research are development of generally applicable methodologies for enzyme immobilization in microstructured elements prepared from different materials; and characterization and optimization of immobilized enzyme microreactors for biocatalytic applications. These can be broken down as follows:

  • Application of (positively charged) silica-binding module (SBM) for reversible immobilization of SBM-enzymes on glass (Year 1 -2)
  • Preparation of a set (minimally 3) SBM-enzymes for further analysis (Year 1.5)
  • Immobilization on microbeads or microchannel walls (Year 2)
  • Enzyme co-immobilization (Year 2 – 3)
  • Extension of immobilization on glass to other materials applied in microfabrication (e.g. organic polymers) (Year 2 – 3)
  • Microreactor operation for continuous biotransformations (Year 1 – 3)
  • Multi-phase enzymatic transformations (Year 3)
Tasks and methodology

The following tasks (with methods allocated) will be addressed in accordance with the objectives specified.

  • Design of enzymes and material surfaces for enzyme immobilization (methods: protein engineering, protein expression, surface modification)
  • Enzyme immobilization and co-immobilization in microreactors (methods: enzyme characterization, protein and enzyme analytics, kinetic analysis)
  • Analysis of enzyme performance in continuously operated microreactors (methods: microreactor set-up and stable operation, analytics)
  • Multi-phase microreactor operation using immobilized enzymes

They are broken down according to Milestones and Deliverables.

  • Comparative analysis of performance microtoolbox
  • Evaluation of immobilization methods suitable for wide range of biocatalysts and for multi-enzyme and chemo-enzyme process conditions completed.
  • Established non-covalent immobilisation methods suitable for IMERs using cationic basic tags and monolithically integrated sol-gels.
  • Biotransformation yields for enzyme variants of TK, TAm and glycosyltransferases established and compared with batch reactions.
  • Microfluidic platform for chemical and enzymatic reactions, amenable for different immobilization methods
  • Biocatalyst screening results using developed reversible immobilization technique completed
  • Report on mass fabrication strategy for selected chemo-enzymatic reactor.