Developing a Protein Scaffolding System for Rapid Enzyme Immobilization and Optimization of Enzyme Functions for Biocatalysis

Guoqiang Zhang, Timothy Johnston, Maureen B. Quin, Claudia Schmidt-Dannert

Research output: Contribution to journalArticlepeer-review

24 Scopus citations

Abstract

Immobilization of enzymes is required for most biocatalytic processes, but chemistries used in enzyme immobilization are limited and can be challenging. Genetically encoded protein-based biomaterials could provide easy-to-use immobilization platforms for biocatalysts. We recently developed a self-assembling protein scaffold that covalently immobilized SpyTagged enzymes by engineering the bacterial microcompartment protein EutM from Salmonella enterica with a SpyCatcher domain. We also identified a range of EutM homologues as robust protein nanostructures with diverse architectures and electrostatic surface properties. In this work, we created a modular immobilization platform with tunable surface properties by developing a toolbox of self-assembling, robust EutM-SpyCatcher scaffolds. Using an alcohol dehydrogenase as model biocatalyst, we show that the scaffolds improve enzyme activity and stability. This work provides a modular, easy-to-use immobilization system that can be tailored for the optimal function of biocatalysts of interest.

Original languageEnglish (US)
Pages (from-to)1867-1876
Number of pages10
JournalACS Synthetic Biology
Volume8
Issue number8
DOIs
StatePublished - Aug 16 2019

Bibliographical note

Funding Information:
TEM imaging of scaffolds was conducted using equipment provided by University Imaging Center, University of Minnesota. This work was supported by funds provided by Defense Threat Reduction Agency Grant HDTRA1-15-0004 and Defense Advanced Research Projects Agency Contract HR0011-17-2-0038. T.J. was supported by funding from a Grand Challenge research award from the University of Minnesota.

Keywords

  • alcohol dehydrogenase
  • biocatalysis
  • enzyme
  • immobilization
  • protein scaffolds
  • synthetic biology

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