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Selection of biocatalysts identified during past screenings.
Only those results are
included that yield commercially interesting products and
are currently not subject to secrecy agreements.
The list is in chronological order (on basis publication date).
Benzylpyrrolidine to N-benzyl-3-
hydroxypyrrolidine
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Reaction: fully regiospecific hydroxylation of N-benzylpyrrolidine in the 3-position yielding either mainly the (S) or (R) enantiomer (strain-dependent) of N-benzyl-3-hydroxypyrrolidine.
Enzymes: range of alkane monooxygenases.
Comments: Both enantiomers of 3-hydroxypyrrolidine are building blocks for different pharmaceuticals
Screening work performed by: Zhi Li, Jan van Beilen and Wouter Duetz
Publications:
Li, Z., Feiten, H.J., Van Beilen, J.B., Duetz, W. and Witholt, B (1999). Preparation of optically active N-benzyl-3-hydroxypyrrolidine by enzymatic hydroxylation. Tetrahedron: Asymmetry 10 :1321-1333.
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Limonene to perillyl alcohol
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Reaction: fully regiospecific hydroxylation of both L- and D-limonene in the 7-position yielding (-) and (+) perillyl alcohol respectively. Activity 10-13 U/g dry wt
Enzymes: range of alkane monooxygenases .
Comments: No activity of archetypal alkane monooxygenase. Also various limonene analogues are substrate to the same hydroxylation of the methyl group. (-) Perillyl alcohol is a promising cancer-drug in phase II trials for various cancers.
Screening work performed by: Ann Fjällman and Wouter Duetz
Publications:
Duetz, W.A., Jourdat, C., Witholt, B. (2001). Process for the preparation of perillyl alcohol, PCT application PCT01103785.0, February 2001
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D-Limonene to (+) trans-carveol
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Reaction: fully regio- and enantio-specific hydroxylation of D-limonene in the 6-position yielding (+) trans-carveol. Actvity 15 U/g dry wt
Enzymes: range of toluene- and naphthalene-dioxygenases.
Comments: Highly unusual monohydroxylation of a non-aromatic compound by an oxygenase whose natural role is the dioxygenation of an aromatic compound.
Screening work performed by: Ann Fjällman and Wouter Duetz
Publications:
Duetz, W.A., Jourdat, C., Witholt, B. (2000). Process for the preparation of trans-carveol. PCT application PCT00124623.0, November 2000
Duetz, W.A., Fjällman, A.H.M., Ren, S., Jourdat, C., and Witholt, B. (2001). Biotransformation of D-limonene to (+) trans-carveol by toluene-grown cells of Rhodococcus opacus PWD4. Appl. Environ. Microbiol., 67: 2829-2832 ( pdf-file, 80 kb)
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p-hydroxyphenylacetic acid to
3,4-dihydroxybenzaldehyde
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Reaction: Combination of oxidations and hydrations leading to the conversion of p-hydroxypheylacetic acid to 3,4 dihydroxybenzaldehyde
Enzymes: Single or multiple oxidase.
Comments: 3,4 dihydroxybenzaldehyde is a fine-chemical that can be used to produce vanillin
Screening work performed by: Kevin O' Connor
Publications:
O'Connor, K.E., Witholt, B., Duetz, W.A. (2001). p-Hydroxyphenylacetic acid metabolism in Pseudomonas putida F6. J. Bacteriol. 183: 928-933 ( pdf-file, 96 kb)
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Diphenylacetylene to
6-phenylacetylene
picolinic Acid
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Reaction: dihydroxylation of one aromatic ring of diphenylacetylene. The cis-glycol formed is further oxidized to a catechol, which is subsequently ring-cleaved, followed by a chemical reaction with ammonium yielding the corresponding picolinic acid
Enzymes: a toluene dioxygenase, a cis-glycol dehydrogenase, and a catechol 2,3-dioxygenase (extradiol ring-cleavage enzyme). The last reaction is non-enzymatical
Comments: Product applicable as end-capping agent for a polymer
Screening work performed by: Jim Spain and Wouter Duetz
Publications:
Spain, J.C., Nishino, S.F., Witholt, B., Tan, L.S., Duetz, W.A. (2003) Production of 6-Phenylacetylene Picolinic Acid from Diphenylacetylene by a Toluene-Degrading Acinetobacter Strain. Appl. Environ. Microbiol. 69:4037-4042
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Enantiospecific reduction of
ethyl 3-keto-4,4,4-trifluorobutyrate
to its chiral alcohols
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Reaction: enantiospecific reduction of a carbonyl group to the corresponding chiral alcohols.
Some enzymes were found to give rise to predominantly the S-form (90% e.e., yield 81%).
Other enzymes were found to give rise to predominantly the R-form (90% e.e., yield 84%).
Enzymes: probably dehydrogenases involved in the degradation of aromatic amino acids
Screening work performed by: Jie Zhang
Publications:
Zhang, J, Duetz, W.A., Witholt, B., and Li, Z. (2004). Rapid identification of new bacterial alcohol dehydrogenases for (R)- and (S)-enantioselective reduction of ß-ketoesters. Chem. Commun. 2120 – 2121
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Enantiospecific reduction of
methyl 3-keto-3-(3’-pyridyl)-propionate
to its chiral alcohols
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Reaction: enantiospecific reduction of a carbonyl group to the corresponding chiral alcohols.
Some enzymes were found to give rise to predominantly the (+)-form (87% e.e., yield 82%).
Some enzymes were found to give rise to predominantly the (-)-form (98% e.e., yield 87%).
Enzymes: probably dehydrogenases involved in the degradation of aromatic amino acids
Screening work performed by: Jie Zhang
Publications:
Zhang, J, Duetz, W.A., Witholt, B., and Li, Z. (2004). Rapid identification of new bacterial alcohol dehydrogenases for (R)- and (S)-enantioselective reduction of ß-ketoesters. Chem. Commun. 2120 – 2121
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