Exploring castellaniella defragrans linalool (De)hydratase-isomerase for enzymatic hydration of alkenes

Matthias Engleder, Monika Müller, Iwona Kaluzna, Daniel Mink, Martin Schürmann, Erich Leitner, Harald Pichler, Anita Emmerstorfer-Augustin

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Acyclic monoterpenes constitute a large and highly abundant class of secondary plant metabolites and are, therefore, attractive low-cost raw materials for the chemical industry. To date, numerous biocatalysts for their transformation are known, giving access to highly sought-after monoterpenoids. In view of the high selectivity associated with many of these reactions, the demand for enzymes generating commercially important target molecules is unabated. Here, linalool (de)hydratase-isomerase (Ldi, EC 4.2.1.127) from Castellaniella defragrans was examined for the regio- and stereoselective hydration of the acyclic monoterpene β-myrcene to (S)-(+)-linalool. Expression of the native enzyme in Escherichia coli allowed for identification of bottlenecks limiting enzyme activity, which were investigated by mutating selected residues implied in enzyme assembly and function. Combining these analyses with the recently published 3D structures of Ldi highlighted the precisely coordinated reduction–oxidation state of two cysteine pairs in correct oligomeric assembly and the catalytic mechanism, respectively. Subcellular targeting studies upon fusion of Ldi to different signal sequences revealed the significance of periplasmic localization of the mature enzyme in the heterologous expression host. This study provides biochemical and mechanistic insight into the hydration of β-myrcene, a nonfunctionalized terpene, and emphasizes its potential for access to scarcely available but commercially interesting tertiary alcohols.

Original languageEnglish
Article number2092
JournalMolecules
Volume24
Issue number11
DOIs
Publication statusPublished - 1 Jun 2019

Keywords

  • Castellaniella defragrans
  • Dehydratase
  • Geraniol
  • Hydratase
  • Isomerase
  • Linalool
  • Monoterpene hydration
  • Myrcene
  • Tertiary alcohol

ASJC Scopus subject areas

  • Analytical Chemistry
  • Chemistry (miscellaneous)
  • Molecular Medicine
  • Pharmaceutical Science
  • Drug Discovery
  • Physical and Theoretical Chemistry
  • Organic Chemistry

Fields of Expertise

  • Human- & Biotechnology

Treatment code (Nähere Zuordnung)

  • Experimental
  • Basic - Fundamental (Grundlagenforschung)

Cooperations

  • BioTechMed-Graz
  • NAWI Graz

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