Characterization and Crystal Structure of a Robust Cyclohexanone Monooxygenase

Overview

Journal Angew Chem Int Ed Engl

Specialty Chemistry

Date 2016 Nov 23

PMID 27873437

Citations 31

Authors

Elvira Romero

J Ruben Gomez Castellanos

Andrea Mattevi

Marco W Fraaije

Affiliations

Soon will be listed here.

Abstract

Cyclohexanone monooxygenase (CHMO) is a promising biocatalyst for industrial reactions owing to its broad substrate spectrum and excellent regio-, chemo-, and enantioselectivity. However, the low stability of many Baeyer-Villiger monooxygenases is an obstacle for their exploitation in industry. Characterization and crystal structure determination of a robust CHMO from Thermocrispum municipale is reported. The enzyme efficiently converts a variety of aliphatic, aromatic, and cyclic ketones, as well as prochiral sulfides. A compact substrate-binding cavity explains its preference for small rather than bulky substrates. Small-scale conversions with either purified enzyme or whole cells demonstrated the remarkable properties of this newly discovered CHMO. The exceptional solvent tolerance and thermostability make the enzyme very attractive for biotechnology.

Citing Articles

Towards biotechnological production of bio-based low molecular weight esters: a patent review.

Zago M, Branduardi P, Serra I RSC Adv. 2024; 14(40):29472-29489.

PMID: 39297040 PMC: 11409443. DOI: 10.1039/d4ra04131c.

Expanding the toolbox of Baeyer-Villiger and flavin monooxygenase biocatalysts for the enantiodivergent green synthesis of sulfoxides.

Wu J, Anselmi S, Carvalho A, Caswell J, Quinn D, Moody T Green Chem. 2024; 26(15):8685-8693.

PMID: 39081496 PMC: 11284623. DOI: 10.1039/d4gc02657h.

Impact of Pulmonary microbiota on lung cancer treatment-related pneumonia.

Zhao M, Hou W, Pu D, Li Z, Tu L, Ow C J Cancer. 2024; 15(14):4503-4512.

PMID: 39006071 PMC: 11242340. DOI: 10.7150/jca.93818.

ALDELE: All-Purpose Deep Learning Toolkits for Predicting the Biocatalytic Activities of Enzymes.

Wang X, Quinn D, Moody T, Huang M J Chem Inf Model. 2024; 64(8):3123-3139.

PMID: 38573056 PMC: 11040732. DOI: 10.1021/acs.jcim.4c00058.

A concise and scalable chemoenzymatic synthesis of prostaglandins.

Yin Y, Wang J, Li J Nat Commun. 2024; 15(1):2523.

PMID: 38514642 PMC: 10957970. DOI: 10.1038/s41467-024-46960-y.

References

Torres Pazmino D, Baas B, Janssen D, Fraaije M . Kinetic mechanism of phenylacetone monooxygenase from Thermobifida fusca. Biochemistry. 2008; 47(13):4082-93. DOI: 10.1021/bi702296k. View

Parra L, Acevedo J, Reetz M . Directed evolution of phenylacetone monooxygenase as an active catalyst for the Baeyer-Villiger conversion of cyclohexanone to caprolactone. Biotechnol Bioeng. 2015; 112(7):1354-64. DOI: 10.1002/bit.25564. View

Vogt G, Woell S, Argos P . Protein thermal stability, hydrogen bonds, and ion pairs. J Mol Biol. 1997; 269(4):631-43. DOI: 10.1006/jmbi.1997.1042. View

Kamerbeek N, Olsthoorn A, Fraaije M, Janssen D . Substrate specificity and enantioselectivity of 4-hydroxyacetophenone monooxygenase. Appl Environ Microbiol. 2003; 69(1):419-26. PMC: 152415. DOI: 10.1128/AEM.69.1.419-426.2003. View

Polyak I, Reetz M, Thiel W . Quantum mechanical/molecular mechanical study on the mechanism of the enzymatic Baeyer-Villiger reaction. J Am Chem Soc. 2012; 134(5):2732-41. DOI: 10.1021/ja2103839. View

Leisch H, Morley K, Lau P . Baeyer-Villiger monooxygenases: more than just green chemistry. Chem Rev. 2011; 111(7):4165-222. DOI: 10.1021/cr1003437. View

Fraaije M, Wu J, Heuts D, van Hellemond E, Lutje Spelberg J, Janssen D . Discovery of a thermostable Baeyer-Villiger monooxygenase by genome mining. Appl Microbiol Biotechnol. 2004; 66(4):393-400. DOI: 10.1007/s00253-004-1749-5. View

Sheng D, Ballou D, MASSEY V . Mechanistic studies of cyclohexanone monooxygenase: chemical properties of intermediates involved in catalysis. Biochemistry. 2001; 40(37):11156-67. DOI: 10.1021/bi011153h. View

Romero E, Gomez Castellanos J, Mattevi A, Fraaije M . Characterization and Crystal Structure of a Robust Cyclohexanone Monooxygenase. Angew Chem Int Ed Engl. 2016; 55(51):15852-15855. PMC: 5213842. DOI: 10.1002/anie.201608951. View

10.

Pechkova E, Sivozhelezov V, Nicolini C . Protein thermal stability: the role of protein structure and aqueous environment. Arch Biochem Biophys. 2007; 466(1):40-8. DOI: 10.1016/j.abb.2007.07.016. View

11.

Yachnin B, McEvoy M, MacCuish R, Morley K, Lau P, Berghuis A . Lactone-bound structures of cyclohexanone monooxygenase provide insight into the stereochemistry of catalysis. ACS Chem Biol. 2014; 9(12):2843-51. DOI: 10.1021/cb500442e. View

12.

Rainey F, Kroppenstedt R, Kempf A, Majazza A, Kutzner H, Stackebrandt E . Thermocrispum gen. nov., a new genus of the order Actinomycetales, and description of Thermocrispum municipale sp. nov. and Thermocrispum agreste sp. nov. Int J Syst Bacteriol. 1995; 45(1):67-77. DOI: 10.1099/00207713-45-1-67. View

13.

van Beek H, Wijma H, Fromont L, Janssen D, Fraaije M . Stabilization of cyclohexanone monooxygenase by a computationally designed disulfide bond spanning only one residue. FEBS Open Bio. 2014; 4:168-74. PMC: 3953729. DOI: 10.1016/j.fob.2014.01.009. View

14.

Zambianchi F, Pasta P, Carrea G, Colonna S, Gaggero N, Woodley J . Use of isolated cyclohexanone monooxygenase from recombinant Escherichia coli as a biocatalyst for Baeyer-Villiger and sulfide oxidations. Biotechnol Bioeng. 2002; 78(5):489-96. DOI: 10.1002/bit.10207. View

15.

Forneris F, Orru R, Bonivento D, Chiarelli L, Mattevi A . ThermoFAD, a Thermofluor-adapted flavin ad hoc detection system for protein folding and ligand binding. FEBS J. 2009; 276(10):2833-40. DOI: 10.1111/j.1742-4658.2009.07006.x. View

16.

Klibanov A . Improving enzymes by using them in organic solvents. Nature. 2001; 409(6817):241-6. DOI: 10.1038/35051719. View

17.

Dudek H, Fink M, Shivange A, Dennig A, Mihovilovic M, Schwaneberg U . Extending the substrate scope of a Baeyer-Villiger monooxygenase by multiple-site mutagenesis. Appl Microbiol Biotechnol. 2013; 98(9):4009-20. DOI: 10.1007/s00253-013-5364-1. View

18.

Opperman D, Reetz M . Towards practical Baeyer-Villiger-monooxygenases: design of cyclohexanone monooxygenase mutants with enhanced oxidative stability. Chembiochem. 2010; 11(18):2589-96. DOI: 10.1002/cbic.201000464. View

19.

Dudek H, de Gonzalo G, Torres Pazmino D, Stepniak P, Wyrwicz L, Rychlewski L . Mapping the substrate binding site of phenylacetone monooxygenase from Thermobifida fusca by mutational analysis. Appl Environ Microbiol. 2011; 77(16):5730-8. PMC: 3165276. DOI: 10.1128/AEM.00687-11. View

20.

Huijbers M, Montersino S, Westphal A, Tischler D, van Berkel W . Flavin dependent monooxygenases. Arch Biochem Biophys. 2013; 544:2-17. DOI: 10.1016/j.abb.2013.12.005. View