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Fungal biocatalysts for labdane diterpene hydroxylation

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Abstract

Labdane diterpenes and their derivatives have shown remarkable biological activities and are useful as chiral building blocks for the synthesis of a variety of bioactive compounds. There is great interest in developing biocatalyst technology to achieve regio- and stereoselective hydroxylation of unactivated C–H bonds in complex natural products, since the functionalization of unactivated C–H bonds generally requires hard reaction conditions and highly reactive oxidizing agents, which are limited regarding the control of regio- and stereoselectivity. Filamentous fungi are efficient biocatalysts capable of catalyzing a wide variety of hydroxylation reactions, and the use of whole cell biocatalysts provides advantages regarding cofactor regeneration and is much less expensive. Therefore, the goal of this study was to select biocatalysts to develop biotransformation processes that can be scalable under mild reaction conditions for hydroxylation of a labdane diterpene, 3β-acetoxy-copalic acid, which contains the trans-decalin moiety and a side chain dienic system appropriate for the preparation of a variety of compounds. Biotransformation processes were carried out and five filamentous fungi were selected as capable of producing hydroxylated diterpenes at positions C-3, C-6, C-7 and C-18 of the trans-decalin moiety and C-13 of the side chain dienic system. Hydroxylation reactions occurred with regio- and stereoselectivity by using some fungi that produced only the 6α, 7α and 13α-hydroxyl derivatives. The chemical structures of the hydroxylated diterpenes were determined from spectrometric and spectroscopic data, and the relative stereochemistry of stereogenic centers was established from coupling constants, by NOE-diff experiments and/or by computational calculations.

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Acknowledgements

The authors are grateful to São Paulo Research Foundation, FAPESP, [Grants 2011/13630-7, 2012/01766-4 and 2016/25201-7], Conselho Nacional de Desenvolvimento Científico e Tecnológico, CNPq, [Grants 306345/2016-1, 306441/2017–9, 313648/2018-2], and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, CAPES, [finance code 001].

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Av. do Café, s/n, Ribeirão Prêto, SP, 14040-903, Brazil

    Tatiane Cruz de Carvalho, Eliane de Oliveira Silva & Niege Araçari Jacometti Cardoso Furtado

  2. Organic Chemistry Department, Chemistry Institute, Federal University of Bahia, Rua Barão de Jeremoabo, s/n, Salvador, BA, 40110-060, Brazil

    Eliane de Oliveira Silva

  3. Nucleus of Research in Sciences and Technology, University of Franca, Franca, SP, Brazil

    Gilberto Augusto Soares, Renato Luis Tame Parreira & Sérgio Ricardo Ambrósio

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  1. Tatiane Cruz de Carvalho
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  2. Eliane de Oliveira Silva
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Cruz de Carvalho, T., de Oliveira Silva, E., Soares, G.A. et al. Fungal biocatalysts for labdane diterpene hydroxylation. Bioprocess Biosyst Eng 43, 1051–1059 (2020). https://doi.org/10.1007/s00449-020-02303-x

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