Engineering TM1459 for Stabilisation against Inactivation by Amino Acid Oxidation

Birgit Grill, Tea Pavkov-Keller, Christoph Grininger, Barbara Darnhofer, Karl Gruber, Mélanie Hall, Helmut Schwab*, Kerstin Steiner*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Oxidative alkene cleavage is a highly interesting reaction to obtain aldehydes and ketones. The Mn-dependent protein TM1459 from Thermotoga maritima can catalyse alkene cleavage of styrene derivatives in the presence of tert-butyl hydroperoxide. Despite the high thermal stability of the enzyme, it gets inactivated during the reaction. The data reported here indicate that auto-oxidation is responsible for the low stability of TM1459 in the oxidative environment required for the alkene cleavage reaction. By targeting the exchange of residues prone to oxidation, this phenomenon was successfully prevented. Importantly, the stability to oxidation conveyed by the amino acid exchanges led to increased enzyme activity. However, the exchanges resulted in slightly modified positions of two of the four metal-binding amino acids, thereby strongly impacting metal binding.

Original languageEnglish
Pages (from-to)596-606
Number of pages11
JournalChemie-Ingenieur-Technik
Volume95
Issue number4
Early online date23 Feb 2023
DOIs
Publication statusPublished - Apr 2023

Keywords

  • Alkene cleavage
  • Amino acid oxidation
  • Cupin
  • Stability

ASJC Scopus subject areas

  • Chemistry(all)
  • Chemical Engineering(all)
  • Industrial and Manufacturing Engineering

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