ACS Publications. Most Trusted. Most Cited. Most Read
Amine Transaminase from Exophiala Xenobiotica—Crystal Structure and Engineering of a Fold IV Transaminase that Naturally Converts Biaryl Ketones

OR SEARCH CITATIONS

My Activity
Publications

Figure 1Loading Img
Download Hi-Res ImageDownload to MS-PowerPointCite This:ACS Catal. 2019, 9, 2, 1140-1148
    Research Article

    Amine Transaminase from Exophiala Xenobiotica—Crystal Structure and Engineering of a Fold IV Transaminase that Naturally Converts Biaryl Ketones
    Click to copy article linkArticle link copied!

    • Aline Telzerow
      Aline Telzerow
      Institute of Molecular Biotechnology, Graz University of Technology, Petersgasse 14, 8010 Graz, Austria
      InnoSyn B. V., Urmonderbaan 22, 6167RD Geleen, The Netherlands
      More by Aline Telzerow
    • Juraj Paris
      Juraj Paris
      Chair of Organic Chemistry I, Faculty of Chemistry, Bielefeld University, Universitätsstraße 25, 33615 Bielefeld, Germany
      Entrechem, S.L., Vivero Ciencias de la Salud, Santo Domingo de Guzmán, s/n, 33011 Oviedo, Spain
      More by Juraj Paris
    • Maria Håkansson
      Maria Håkansson
      SARomics Biostructures AB, Medicon Village, Scheelevägen 2, 22381 Lund, Sweden
      More by Maria Håkansson
    • Javier González-Sabín
      Javier González-Sabín
      Entrechem, S.L., Vivero Ciencias de la Salud, Santo Domingo de Guzmán, s/n, 33011 Oviedo, Spain
      More by Javier González-Sabín
      Orcidhttp://orcid.org/0000-0003-3764-4750
    • Nicolás Ríos-Lombardía
      Nicolás Ríos-Lombardía
      Entrechem, S.L., Vivero Ciencias de la Salud, Santo Domingo de Guzmán, s/n, 33011 Oviedo, Spain
      More by Nicolás Ríos-Lombardía
    • Martin Schürmann
      Martin Schürmann
      InnoSyn B. V., Urmonderbaan 22, 6167RD Geleen, The Netherlands
      More by Martin Schürmann
    • Harald Gröger
      Harald Gröger
      Chair of Organic Chemistry I, Faculty of Chemistry, Bielefeld University, Universitätsstraße 25, 33615 Bielefeld, Germany
      More by Harald Gröger
      Orcidhttp://orcid.org/0000-0001-8582-2107
    • Francisco Morís
      Francisco Morís
      Entrechem, S.L., Vivero Ciencias de la Salud, Santo Domingo de Guzmán, s/n, 33011 Oviedo, Spain
      More by Francisco Morís
    • Robert Kourist
      Robert Kourist
      Institute of Molecular Biotechnology, Graz University of Technology, Petersgasse 14, 8010 Graz, Austria
      More by Robert Kourist
    • Helmut Schwab
      Helmut Schwab
      Institute of Molecular Biotechnology, Graz University of Technology, Petersgasse 14, 8010 Graz, Austria
      More by Helmut Schwab
    • Kerstin Steiner*
      Kerstin Steiner
      Institute of Molecular Biotechnology, Graz University of Technology, Petersgasse 14, 8010 Graz, Austria
      *E-mail: [email protected]
      More by Kerstin Steiner
      Orcidhttp://orcid.org/0000-0002-9037-3972
    Other Access OptionsSupporting Information (1)

    ACS Catalysis

    Cite this: ACS Catal. 2019, 9, 2, 1140–1148
    Click to copy citationCitation copied!
    https://doi.org/10.1021/acscatal.8b04524
    Published December 20, 2018
    Copyright © 2018 American Chemical Society
    Request reuse permissions

    Abstract

    Click to copy section linkSection link copied!
    Abstract Image

    Amine transaminases are frequently used for the production of chiral amines starting from prochiral ketones. These amines can be applied as active pharmaceutical ingredients or drug precursors. However, there are still limitations to the use of amine transaminases when it comes to bulky ketone substrates, such as biaryl ketones. Using data mining, an (R)-selective amine transaminase from Exophiala xenobiotica was identified which naturally converts biaryl ketone substrates to the corresponding amines with up to 85% conversion and excellent enantioselectivity (>99% ee). Its protein crystal structure was obtained with a resolution of 1.52 Å, which enabled us to explain this interesting substrate acceptance. Structure-guided protein engineering resulted in a quintuple variant with increased stability. Moreover, the amino acid exchange T273S increased the activity and broadened the substrate scope, enabling conversions of various biaryl ketones with up to >99%. A preparative biotransformation of 1-(4-(pyridin-3-yl)phenyl)ethenone at 75 mM (15 g/L) resulted in 96% of isolated yield of the respective amine.

    Copyright © 2018 American Chemical Society

    Subjects

    what are subjects

    Keywords

    what are keywords

    Read this article

    To access this article, please review the available access options below.

    Get instant access

    Purchase Access

    Read this article for 48 hours. Check out below using your ACS ID or as a guest.

    Recommended

    Access through Your Institution

    You may have access to this article through your institution.

    Your institution does not have access to this content. Add or change your institution or let them know you’d like them to include access.

    Supporting Information

    Click to copy section linkSection link copied!

    The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acscatal.8b04524.

    • Conserved motifs of fold IV PLP-dependent enzymes, list of primers, protein crystallography data; additional experimental procedures and data for protein engineering; omit electron density of PLP in the structure, biocatalyst formulation study, transamination of 1i using isopropylamine as amino donor, substrate loading studies, enzyme loading studies; detailed results of enzymatic transamination of ketones 1a–j; assignments of the absolute configurations, spectroscopic data of amines 2a–b, e-j; HPLC analytical data; copies of chiral-HPLC chromatograms; copies of NMR spectra (PDF)

    Terms & Conditions

    Most electronic Supporting Information files are available without a subscription to ACS Web Editions. Such files may be downloaded by article for research use (if there is a public use license linked to the relevant article, that license may permit other uses). Permission may be obtained from ACS for other uses through requests via the RightsLink permission system: http://pubs.acs.org/page/copyright/permissions.html.

    Cited By

    Click to copy section linkSection link copied!

    This article is cited by 36 publications.

    1. Sam Mathew, Dominik Renn, Magnus Rueping. Advances in One-Pot Chiral Amine Synthesis Enabled by Amine Transaminase Cascades: Pushing the Boundaries of Complexity. ACS Catalysis 2023, 13 (8) , 5584-5598. https://doi.org/10.1021/acscatal.3c00555
    2. Fulong Li, Yan Du, Youxiang Liang, Yuwen Wei, Yukun Zheng, Huimin Yu. Redesigning an (R)-Selective Transaminase for the Efficient Synthesis of Pharmaceutical N-Heterocyclic Amines. ACS Catalysis 2023, 13 (1) , 422-432. https://doi.org/10.1021/acscatal.2c05177
    3. Qinglong Meng, Carlos Ramírez-Palacios, Nikolas Capra, Mattijs E. Hooghwinkel, Sebastian Thallmair, Henriëtte J. Rozeboom, Andy-Mark W. H. Thunnissen, Hein J. Wijma, Siewert J. Marrink, Dick B. Janssen. Computational Redesign of an ω-Transaminase from Pseudomonas jessenii for Asymmetric Synthesis of Enantiopure Bulky Amines. ACS Catalysis 2021, 11 (17) , 10733-10747. https://doi.org/10.1021/acscatal.1c02053
    4. Moritz Voss, Chao Xiang, Jérémy Esque, Alberto Nobili, Marian J. Menke, Isabelle André, Matthias Höhne, Uwe T. Bornscheuer. Creation of (R)-Amine Transaminase Activity within an α-Amino Acid Transaminase Scaffold. ACS Chemical Biology 2020, 15 (2) , 416-424. https://doi.org/10.1021/acschembio.9b00888
    5. Juraj Paris, Aline Telzerow, Nicolás Ríos-Lombardía, Kerstin Steiner, Helmut Schwab, Francisco Morís, Harald Gröger, Javier González-Sabín. Enantioselective One-Pot Synthesis of Biaryl-Substituted Amines by Combining Palladium and Enzyme Catalysis in Deep Eutectic Solvents. ACS Sustainable Chemistry & Engineering 2019, 7 (5) , 5486-5493. https://doi.org/10.1021/acssuschemeng.8b06715
    6. Xiao Gao, Wenhe Zhang, Xiaowei Wei, Lu Zhao, Changli Che, Zhuobing Zhang, Hongli Wei, Bin Qin, Weidong Liu, Xian Jia, Song You. Structure-guided engineering an (R)-transaminase from Mycobacterium neoaurum for efficient synthesis of chiral N-heterocyclic amines. International Journal of Biological Macromolecules 2025, 287 , 138591. https://doi.org/10.1016/j.ijbiomac.2024.138591
    7. Stefania Cananà, Federica De Nardi, Marco Blangetti, Stefano Parisotto, Cristina Prandi. Biocatalysis in Non‐Conventional Media: Unlocking the Potential for Sustainable Chiral Amine Synthesis. Chemistry – A European Journal 2024, https://doi.org/10.1002/chem.202304364
    8. Xiaoping Yue, Yitong Li, Mankun Wei, Yu Duan, Lin Yang, Fen-Er Chen. Rational redesign of the loop dynamics of carbonyl reductase LfSDR1 to improve the stereoselectivity for asymmetric synthesis of bulky chiral alcohols. International Journal of Biological Macromolecules 2024, 274 , 133345. https://doi.org/10.1016/j.ijbiomac.2024.133345
    9. J. M. Carceller, K. S. Arias, M. J. Climent, S. Iborra, A. Corma. One-pot chemo- and photo-enzymatic linear cascade processes. Chemical Society Reviews 2024, 53 (15) , 7875-7938. https://doi.org/10.1039/D3CS00595J
    10. Quan Luo, Guan Zhou, Zhongxia Li, Jiangpeng Dong, Hang Zhao, Huifang Xu, Xuefeng Lu. ω-transaminase-catalyzed synthesis of (R)-2-(1-aminoethyl)-4-fluorophenol, a chiral intermediate of novel anti-tumor drugs. Enzyme and Microbial Technology 2024, 175 , 110406. https://doi.org/10.1016/j.enzmictec.2024.110406
    11. Tingting Li, Hai Zhu, Weiwei Jia, Xia Tian, Ziwen Xu, Jiang Zhu, Wencheng Liu, Yang Cao. Identification, characterization and application of M16AT, a new organic solvent–tolerant ( R )–enantio–selective type IV amine transaminase from Mycobacterium sp. ACS1612. ChemBioChem 2024, 363 https://doi.org/10.1002/cbic.202300812
    12. Sofia A. Shilova, Maria G. Khrenova, Ilya O. Matyuta, Alena Y. Nikolaeva, Tatiana V. Rakitina, Natalia L. Klyachko, Mikhail E. Minyaev, Konstantin M. Boyko, Vladimir O. Popov, Ekaterina Yu. Bezsudnova. To the Understanding of Catalysis by D-Amino Acid Transaminases: A Case Study of the Enzyme from Aminobacterium colombiense. Molecules 2023, 28 (5) , 2109. https://doi.org/10.3390/molecules28052109
    13. Lin Yang, Kaiyue Zhang, Meng Xu, Youyu Xie, Xiangqi Meng, Hualei Wang, Dongzhi Wei. Mechanism‐Guided Computational Design of ω‐Transaminase by Reprograming of High‐Energy‐Barrier Steps. Angewandte Chemie International Edition 2022, 61 (52) https://doi.org/10.1002/anie.202212555
    14. Lin Yang, Kaiyue Zhang, Meng Xu, Youyu Xie, Xiangqi Meng, Hualei Wang, Dongzhi Wei. Mechanism‐Guided Computational Design of ω‐Transaminase by Reprograming of High‐Energy‐Barrier Steps. Angewandte Chemie 2022, 134 (52) https://doi.org/10.1002/ange.202212555
    15. Guozeng Wang, Zhihao Jiang, Qing Xiao, Chang Jiang, Xian'ai Shi. Visible spectrophotometric assay for characterization of ω-transaminases. Analytical Biochemistry 2022, 658 , 114933. https://doi.org/10.1016/j.ab.2022.114933
    16. Qinglong Meng, Carlos Ramírez-Palacios, Hein J. Wijma, Dick B. Janssen. Protein engineering of amine transaminases. Frontiers in Catalysis 2022, 2 https://doi.org/10.3389/fctls.2022.1049179
    17. Tingting Wang, Changmeng Liu, Dong Xu, Jiaxi Xu, Zhanhui Yang. Iridium-Catalyzed and pH-Dependent Reductions of Nitroalkenes to Ketones. Molecules 2022, 27 (22) , 7822. https://doi.org/10.3390/molecules27227822
    18. Yuriy V. Sheludko, Sjoerd Slagman, Samantha Gittings, Simon J. Charnock, Henrik Land, Per Berglund, Wolf‐Dieter Fessner. Enantioselective Synthesis of Pharmaceutically Relevant Bulky Arylbutylamines Using Engineered Transaminases. Advanced Synthesis & Catalysis 2022, 364 (17) , 2972-2981. https://doi.org/10.1002/adsc.202200403
    19. Amol D. Pagar, Hyunwoo Jeon, Taresh P. Khobragade, Sharad Sarak, Pritam Giri, Seonga Lim, Tae Hyeon Yoo, Byoung Joon Ko, Hyungdon Yun. Non-Canonical Amino Acid-Based Engineering of (R)-Amine Transaminase. Frontiers in Chemistry 2022, 10 https://doi.org/10.3389/fchem.2022.839636
    20. Fulong Li, Youxiang Liang, Yuwen Wei, Yukun Zheng, Yan Du, Huimin Yu. Biochemical and Structural Characterization of an ( R )‐Selective Transaminase in the Asymmetric Synthesis of Chiral Hydroxy Amines. Advanced Synthesis & Catalysis 2021, 363 (19) , 4582-4589. https://doi.org/10.1002/adsc.202100636
    21. Yunting Liu, Pengbo Liu, Shiqi Gao, Zihan Wang, Pengqian Luan, Javier González-Sabín, Yanjun Jiang. Construction of chemoenzymatic cascade reactions for bridging chemocatalysis and Biocatalysis: Principles, strategies and prospective. Chemical Engineering Journal 2021, 420 , 127659. https://doi.org/10.1016/j.cej.2020.127659
    22. Jesús Albarrán‐Velo, Vicente Gotor‐Fernández, Iván Lavandera. Markovnikov Wacker‐Tsuji Oxidation of Allyl(hetero)arenes and Application in a One‐Pot Photo‐Metal‐Biocatalytic Approach to Enantioenriched Amines and Alcohols. Advanced Synthesis & Catalysis 2021, 363 (16) , 4096-4108. https://doi.org/10.1002/adsc.202100351
    23. Ekaterina Yu. Bezsudnova, Alena Yu. Nikolaeva, Alina K. Bakunova, Tatiana V. Rakitina, Dmitry A. Suplatov, Vladimir O. Popov, Konstantin M. Boyko, . Probing the role of the residues in the active site of the transaminase from Thermobaculum terrenum. PLOS ONE 2021, 16 (7) , e0255098. https://doi.org/10.1371/journal.pone.0255098
    24. Yue Sun, Elia Calderini, Robert Kourist. A Reconstructed Common Ancestor of the Fatty Acid Photo‐decarboxylase Clade Shows Photo‐decarboxylation Activity and Increased Thermostability. ChemBioChem 2021, 22 (10) , 1833-1840. https://doi.org/10.1002/cbic.202000851
    25. Andrea M. Chánique, Nicole Dimos, Ivana Drienovská, Elia Calderini, Mónica P. Pantín, Carl P. O. Helmer, Michael Hofer, Volker Sieber, Loreto P. Parra, Bernhard Loll, Robert Kourist. A Structural View on the Stereospecificity of Plant Borneol‐Type Dehydrogenases. ChemCatChem 2021, 13 (9) , 2262-2277. https://doi.org/10.1002/cctc.202100110
    26. Robert Kourist, Javier González‐Sabín. Compartmentalization in Biocatalysis. 2021, 89-112. https://doi.org/10.1002/9783527824465.ch4
    27. Aline Telzerow, Juraj Paris, Maria Håkansson, Javier González‐Sabín, Nicolas Ríos‐Lombardía, Harald Gröger, Francisco Morís, Martin Schürmann, Helmut Schwab, Kerstin Steiner. Expanding the Toolbox of R ‐Selective Amine Transaminases by Identification and Characterization of New Members. ChemBioChem 2021, 22 (7) , 1232-1242. https://doi.org/10.1002/cbic.202000692
    28. Javier González-Sabín. Enzymatic Cascade Reactions in Non-Conventional Media. 2021, 165-178. https://doi.org/10.1007/978-3-030-65718-5_10
    29. Ágnes Lakó, Zsófia Molnár, Ricardo Mendonça, László Poppe. Transaminase-mediated synthesis of enantiopure drug-like 1-(3′,4′-disubstituted phenyl)propan-2-amines. RSC Advances 2020, 10 (67) , 40894-40903. https://doi.org/10.1039/D0RA08134E
    30. Christian M. Heckmann, Louise J. Gourlay, Beatriz Dominguez, Francesca Paradisi. An (R)-Selective Transaminase From Thermomyces stellatus: Stabilizing the Tetrameric Form. Frontiers in Bioengineering and Biotechnology 2020, 8 https://doi.org/10.3389/fbioe.2020.00707
    31. Xinxing Gao, Xin Zhang, Nianqing Zhu, Yi Mou, Hailing Zhang, Xin Liu, Pinghe Wei. Reshaping the substrate binding region of (R)-selective ω-transaminase for asymmetric synthesis of (R)-3-amino-1-butanol. Applied Microbiology and Biotechnology 2020, 104 (9) , 3959-3969. https://doi.org/10.1007/s00253-020-10539-6
    32. Robert Kourist, Javier González‐Sabín. Non‐Conventional Media as Strategy to Overcome the Solvent Dilemma in Chemoenzymatic Tandem Catalysis. ChemCatChem 2020, 12 (7) , 1903-1912. https://doi.org/10.1002/cctc.201902192
    33. Ekaterina Yu. Bezsudnova, Vladimir O. Popov, Konstantin M. Boyko. Structural insight into the substrate specificity of PLP fold type IV transaminases. Applied Microbiology and Biotechnology 2020, 104 (6) , 2343-2357. https://doi.org/10.1007/s00253-020-10369-6
    34. Henrik Land, Federica Ruggieri, Anna Szekrenyi, Wolf‐Dieter Fessner, Per Berglund. Engineering the Active Site of an ( S )‐Selective Amine Transaminase for Acceptance of Doubly Bulky Primary Amines. Advanced Synthesis & Catalysis 2020, 362 (4) , 812-821. https://doi.org/10.1002/adsc.201901252
    35. Federica Ruggieri, Jonatan C. Campillo-Brocal, Shan Chen, Maria S. Humble, Björn Walse, Derek T. Logan, Per Berglund. Insight into the dimer dissociation process of the Chromobacterium violaceum (S)-selective amine transaminase. Scientific Reports 2019, 9 (1) https://doi.org/10.1038/s41598-019-53177-3
    36. Robert A. Hill, Andrew Sutherland. Hot off the Press. Natural Product Reports 2019, 36 (4) , 556-560. https://doi.org/10.1039/C9NP90010A
    Download PDF
    Go to ACS Catalysis
    Get e-Alerts
    Get e-Alerts

    ACS Catalysis

    Cite this: ACS Catal. 2019, 9, 2, 1140–1148
    Click to copy citationCitation copied!
    https://doi.org/10.1021/acscatal.8b04524
    Published December 20, 2018
    Copyright © 2018 American Chemical Society
    Request reuse permissions

    Article Views

    2374

    Altmetric

    -

    Citations

    36
    Learn about these metrics

    Article Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.

    Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.

    The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated.