Biocatalytic Production of Perillyl Alcohol from Limonene by Using a Novel Mycobacterium sp. Cytochrome P450 Alkane Hydroxylase Expressed in Pseudomonas putida
- van Beilen, Jan B. Institute of Biotechnology, ETH Hönggerberg, Zürich, Switzerland
- Holtackers, René Institute of Biotechnology, ETH Hönggerberg, Zürich, Switzerland
- Lüscher, Daniel Institute of Biotechnology, ETH Hönggerberg, Zürich, Switzerland
- Bauer, Ulrich Institute of Biotechnology, ETH Hönggerberg, Zürich, Switzerland
- Witholt, Bernard Institute of Biotechnology, ETH Hönggerberg, Zürich, Switzerland
- Duetz, Wouter A. Enzyscreen BV, Leiden, The Netherlands
Published in:
- Applied and Environmental Microbiology. - American Society for Microbiology. - 2005, vol. 71, no. 4, p. 1737-1744
English
ABSTRACT
A number of oxygenated monoterpenes present at low concentrations in plant oils have anticarcinogenic properties. One of the most promising compounds in this respect is (−)-perillyl alcohol. Since this natural product is present only at low levels in a few plant oils, an alternative, synthetic source is desirable. Screening of 1,800 bacterial strains showed that many alkane degraders were able to specifically hydroxylate l -limonene in the 7 position to produce enantiopure (−)-perillyl alcohol. The oxygenase responsible for this was purified from the best-performing wild-type strain, Mycobacterium sp. strain HXN-1500. By using N-terminal sequence information, a 6.2-kb ApaI fragment was cloned, which encoded a cytochrome P450, a ferredoxin, and a ferredoxin reductase. The three genes were successfully coexpressed in Pseudomonas putida by using the broad-host-range vector pCom8, and the recombinant converted limonene to perillyl alcohol with a specific activity of 3 U/g (dry weight) of cells. The construct was subsequently used in a 2-liter bioreactor to produce perillyl alcohol on a scale of several grams.
- Language
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- English
- Open access status
- bronze
- Identifiers
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- DOI 10.1128/aem.71.4.1737-1744.2005
- ISSN 0099-2240
- Persistent URL
- https://sonar.ch/global/documents/250445
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