Engineering Thermostability in Artificial Metalloenzymes to Increase Catalytic Activity

Megan Doble, Lorenz Obrecht, Henk-Jan Joosten, Misun Lee, Henriette J. Rozeboom, Emma Branigan, James H. Naismith, Dick B. Janssen, Amanda G. Jarvis*, Paul C. J. Kamer

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Protein engineering has shown widespread use in improving the industrial application of enzymes and broadening the conditions they are able to operate under by increasing their thermostability and solvent tolerance. Here, we show that protein engineering can be used to increase the thermostability of an artificial metalloenzyme. Thermostable variants of the human steroid carrier protein 2L, modified to bind a metal catalyst, were created by rational design using structural data and a 3DM database. These variants were tested to identify mutations that enhanced the stability of the protein scaffold, and a significant increase in melting temperature was observed with a number of modified metalloenzymes. The ability to withstand higher reaction temperatures resulted in an increased activity in the hydroformylation of 1-octene, with more than fivefold improvement in turnover number, whereas the selectivity for linear aldehyde remained high up to 80%.

Original languageEnglish
Pages (from-to)3620-3627
Number of pages8
JournalACS Catalysis
Volume11
Issue number6
DOIs
Publication statusPublished - 19-Mar-2021

Keywords

  • artificial metalloenzyme
  • hydroformylation
  • bioengineering
  • protein thermostability
  • biocatalysis
  • bioinformatics

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