Engineering of a borneol dehydrogenase from P. putida for the enzymatic resolution of camphor

Michael Hofer*, Julia Diener, Benjamin Begander, Robert Kourist, Volker Sieber

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


Abstract: Several thousand different terpenoid structures are known so far, and many of them are interesting for applications as pharmaceuticals, flavors, fragrances, biofuels, insecticides, or fine chemical intermediates. One prominent example is camphor, which has been utilized since ancient times in medical applications. Especially (−)-camphor is gaining more and more interest for pharmaceutical applications. Hence, a commercial reliable source is needed. The natural sources for (−)-camphor are limited, and the oxidation of precious (−)-borneol would be too costly. Hence, synthesis of (−)-camphor from renewable alpha-pinene would be an inexpensive alternative. As the currently used route for the conversion of alpha-pinene to camphor produces a mixture of both enantiomers, preferably catalytic methods for the separation of this racemate are demanded to yield enantiopure camphor. Enzymatic kinetic resolution is a sustainable way to solve this challenge but requires suitable enzymes. In this study, the first borneol dehydrogenase from Pseudomonas sp. ATCC 17453, capable of catalyzing the stereoselective reduction of camphor, was examined. By using a targeted enzyme engineering approach, enantioselective enzyme variants were created with E-values > 100. The best variant was used for the enzymatic kinetic resolution of camphor racemate, yielding 79% of (−)-camphor with an ee of > 99%. Key points: • Characterization of a novel borneol dehydrogenase (BDH) from P. putida. • Development of enantioselective BDH variants for the reduction of camphor. • Enzymatic kinetic resolution of camphor with borneol dehydrogenase. Graphical abstract: [Figure not available: see fulltext.]

Original languageEnglish
Pages (from-to)3159-3167
Number of pages9
JournalApplied Microbiology and Biotechnology
Issue number8
Publication statusPublished - Apr 2021


  • Borneol dehydrogenase
  • Camphor
  • Enzymatic resolution
  • Racemate

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology


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