10. Fusion to a Pull-Down Module

Designing Enzymes to Form Biocatalytically Active Insoluble Aggregates

  1. Silvia Maria Doglia and
  2. Marina Lotti
  1. Bernd Nidetzky

Published Online: 11 APR 2014

DOI: 10.1002/9781118845363.ch10

Protein Aggregation in Bacteria: Functional and Structural Properties of Inclusion Bodies in Bacterial Cells

Protein Aggregation in Bacteria: Functional and Structural Properties of Inclusion Bodies in Bacterial Cells

How to Cite

Nidetzky, B. (2014) Fusion to a Pull-Down Module, in Protein Aggregation in Bacteria: Functional and Structural Properties of Inclusion Bodies in Bacterial Cells (eds S. M. Doglia and M. Lotti), John Wiley & Sons, Inc., Hoboken, NJ. doi: 10.1002/9781118845363.ch10

Publication History

  1. Published Online: 11 APR 2014
  2. Published Print: 18 APR 2014

ISBN Information

Print ISBN: 9781118448526

Online ISBN: 9781118845363

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Keywords:

  • biocatalyst;
  • cross-linked enzyme aggregates (CLEAs);
  • encapsulation;
  • enzymes;
  • insoluble protein aggregate;
  • pull-down module (PDM)

Summary

This chapter describes the recent developments in the preparation and application of insoluble enzyme, dicussing different methods of carrier-free immobilization. It focuses on molecular approaches by protein design, where an aggregation-prone or self-assembling structural element, referred to as a pull-down module (PDM), is appended to the enzyme of interest. Recombinant production of the PDM–enzyme fusion should thus yield an insoluble protein aggregate that can be recovered easily, processed further if needed, and then used as an insoluble biocatalyst. Magnetic cross-linked enzyme aggregates (CLEAs) might be applied in combination with fluidized beds, using magnetic field for retention of the insoluble enzyme. The use of CLEAs is widely adopted by the biocatalysis community. Encapsulation or any other form of processing postaggregation may eventually be necessary cases to obtain mechanically stable biocatalysts. New protocols, such as preparation of composites from protein aggregates and other materials, makes an interesting option for processing postaggregation.