Structural and functional insights into asymmetric enzymatic dehydration of alkenols

Bettina M Nestl; Christopher Geinitz; Stephanie Popa; Sari Rizek; Robert J Haselbeck; Rosary Stephen; Michael A Noble; Max-Philipp Fischer; Erik C Ralph; Hoi Ting Hau; Henry Man; Muhiadin Omar; Johan P Turkenburg; Stephen van Dien; Stephanie J Culler; Gideon Grogan; Bernhard Hauer

doi:10.1038/nchembio.2271

Structural and functional insights into asymmetric enzymatic dehydration of alkenols

Bettina M Nestl, Christopher Geinitz, Stephanie Popa, Sari Rizek, Robert J Haselbeck, Rosary Stephen, Michael A Noble, Max-Philipp Fischer, Erik C Ralph, Hoi Ting Hau, Henry Man, Muhiadin Omar, Johan P Turkenburg, Stephen van Dien, Stephanie J Culler, Gideon Grogan, Bernhard Hauer

Chemistry

Research output: Contribution to journal › Article › peer-review

Abstract

The asymmetric dehydration of alcohols is an important process for the direct synthesis of alkenes. We report the structure and substrate specificity of the bifunctional linalool dehydratase isomerase (LinD) from the bacterium Castellaniella defragrans that catalyzes in nature the hydration of β-myrcene to linalool and the subsequent isomerization to geraniol. Enzymatic kinetic resolutions of truncated and elongated aromatic and aliphatic tertiary alcohols (C5-C15) that contain a specific signature motif demonstrate the broad substrate specificity of LinD. The three-dimensional structure of LinD from Castellaniella defragrans revealed a pentamer with active sites at the protomer interfaces. Furthermore, the structure of LinD in complex with the product geraniol provides initial mechanistic insights into this bifunctional enzyme. Site-directed mutagenesis confirmed active site amino acid residues essential for its dehydration and isomerization activity. These structural and mechanistic insights facilitate the development of hydrating catalysts, enriching the toolbox for novel bond-forming biocatalysis.

Original language	English
Pages (from-to)	275-281
Number of pages	7
Journal	NATURE CHEMICAL BIOLOGY
Volume	13
Issue number	3
Early online date	9 Jan 2017
DOIs	https://doi.org/10.1038/nchembio.2271
Publication status	Published - 1 Mar 2017

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© 2017, The publisher.This is an author-produced version of the published paper. Uploaded in accordance with the publisher’s self-archiving policy. Further copying may not be permitted; contact the publisher for details

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Nestl, B. M., Geinitz, C., Popa, S., Rizek, S., Haselbeck, R. J., Stephen, R., Noble, M. A., Fischer, M-P., Ralph, E. C., Hau, H. T., Man, H., Omar, M., Turkenburg, J. P., van Dien, S., Culler, S. J., Grogan, G., & Hauer, B. (2017). Structural and functional insights into asymmetric enzymatic dehydration of alkenols. NATURE CHEMICAL BIOLOGY, 13(3), 275-281. https://doi.org/10.1038/nchembio.2271

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abstract = "The asymmetric dehydration of alcohols is an important process for the direct synthesis of alkenes. We report the structure and substrate specificity of the bifunctional linalool dehydratase isomerase (LinD) from the bacterium Castellaniella defragrans that catalyzes in nature the hydration of β-myrcene to linalool and the subsequent isomerization to geraniol. Enzymatic kinetic resolutions of truncated and elongated aromatic and aliphatic tertiary alcohols (C5-C15) that contain a specific signature motif demonstrate the broad substrate specificity of LinD. The three-dimensional structure of LinD from Castellaniella defragrans revealed a pentamer with active sites at the protomer interfaces. Furthermore, the structure of LinD in complex with the product geraniol provides initial mechanistic insights into this bifunctional enzyme. Site-directed mutagenesis confirmed active site amino acid residues essential for its dehydration and isomerization activity. These structural and mechanistic insights facilitate the development of hydrating catalysts, enriching the toolbox for novel bond-forming biocatalysis.",

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Nestl, BM, Geinitz, C, Popa, S, Rizek, S, Haselbeck, RJ, Stephen, R, Noble, MA, Fischer, M-P, Ralph, EC, Hau, HT, Man, H, Omar, M, Turkenburg, JP, van Dien, S, Culler, SJ, Grogan, G & Hauer, B 2017, 'Structural and functional insights into asymmetric enzymatic dehydration of alkenols', NATURE CHEMICAL BIOLOGY, vol. 13, no. 3, pp. 275-281. https://doi.org/10.1038/nchembio.2271

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AU - Geinitz, Christopher

AU - Popa, Stephanie

AU - Rizek, Sari

AU - Haselbeck, Robert J

AU - Stephen, Rosary

AU - Noble, Michael A

AU - Fischer, Max-Philipp

AU - Ralph, Erik C

AU - Hau, Hoi Ting

AU - Man, Henry

AU - Omar, Muhiadin

AU - Turkenburg, Johan P

AU - van Dien, Stephen

AU - Culler, Stephanie J

AU - Grogan, Gideon

AU - Hauer, Bernhard

N1 - © 2017, The publisher.This is an author-produced version of the published paper. Uploaded in accordance with the publisher’s self-archiving policy. Further copying may not be permitted; contact the publisher for details

PY - 2017/3/1

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N2 - The asymmetric dehydration of alcohols is an important process for the direct synthesis of alkenes. We report the structure and substrate specificity of the bifunctional linalool dehydratase isomerase (LinD) from the bacterium Castellaniella defragrans that catalyzes in nature the hydration of β-myrcene to linalool and the subsequent isomerization to geraniol. Enzymatic kinetic resolutions of truncated and elongated aromatic and aliphatic tertiary alcohols (C5-C15) that contain a specific signature motif demonstrate the broad substrate specificity of LinD. The three-dimensional structure of LinD from Castellaniella defragrans revealed a pentamer with active sites at the protomer interfaces. Furthermore, the structure of LinD in complex with the product geraniol provides initial mechanistic insights into this bifunctional enzyme. Site-directed mutagenesis confirmed active site amino acid residues essential for its dehydration and isomerization activity. These structural and mechanistic insights facilitate the development of hydrating catalysts, enriching the toolbox for novel bond-forming biocatalysis.

AB - The asymmetric dehydration of alcohols is an important process for the direct synthesis of alkenes. We report the structure and substrate specificity of the bifunctional linalool dehydratase isomerase (LinD) from the bacterium Castellaniella defragrans that catalyzes in nature the hydration of β-myrcene to linalool and the subsequent isomerization to geraniol. Enzymatic kinetic resolutions of truncated and elongated aromatic and aliphatic tertiary alcohols (C5-C15) that contain a specific signature motif demonstrate the broad substrate specificity of LinD. The three-dimensional structure of LinD from Castellaniella defragrans revealed a pentamer with active sites at the protomer interfaces. Furthermore, the structure of LinD in complex with the product geraniol provides initial mechanistic insights into this bifunctional enzyme. Site-directed mutagenesis confirmed active site amino acid residues essential for its dehydration and isomerization activity. These structural and mechanistic insights facilitate the development of hydrating catalysts, enriching the toolbox for novel bond-forming biocatalysis.

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Structural and functional insights into asymmetric enzymatic dehydration of alkenols

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Gideon James Grogan

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Structural and functional insights into asymmetric enzymatic dehydration of alkenols

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Gideon James Grogan

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