Article
Biotechnology & Applied Microbiology
Martha Sophia Smit, Mpeyake Jacob Maseme, Jacqueline van Marwijk, Jasmin Cara Aschenbrenner, Diederik Johannes Opperman
Summary: Cytochrome P450 monooxygenases (CYP450s) are crucial in the synthesis and breakdown of secondary metabolites in eukaryotes, especially in plants and fungi. CYP505E3 from Aspergillus terreus demonstrates remarkable omega-7 hydroxylase activity, making it a promising catalyst for delta-dodecalactone synthesis. Other CYP505Es also display similar activity, but their occurrence is limited to certain species like Aspergillus and Penicillium.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2023)
Article
Biochemistry & Molecular Biology
Yuri D. Ivanov, Amir Taldaev, Andrey V. Lisitsa, Elena A. Ponomarenko, Alexander I. Archakov
Summary: The study predicted the three-dimensional structures of CYP102A1/WT proteins and their mutant forms using AlphaFold programs. The structures of monomers in the homodimer were found to be consistent with experimental structures, while the predicted structures of isolated monomers were different. Significant differences were observed in the structure of monomers compared to homodimers for the mutant forms. Comparative analysis showed a correlation between the electron transfer rate constants and the distances between prosthetic groups.
Review
Engineering, Chemical
Bekir Engin Eser, Yan Zhang, Li Zong, Zheng Guo
Summary: Cytochrome P450s (CYPs) are widely distributed in all kingdoms of life, with self-sufficient CYPs being particularly notable for their efficient catalytic capabilities and simpler systems, playing important roles in various synthetic and degradative pathways.
CHINESE JOURNAL OF CHEMICAL ENGINEERING
(2021)
Article
Chemistry, Physical
Cheri Louise Jacobs, Rodolpho do Aido-Machado, Carmien Tolmie, Martha Sophia Smit, Diederik Johannes Opperman
Summary: Selective oxyfunctionalization of non-activated C-H bonds is a significant challenge in synthetic chemistry. In this study, the biocatalytic hydroxylation of medium-chain n-alkanes by the bacterial enzyme CYP153A71 was characterized. The enzyme was found to convert 1-alkanols to aldehydes, fatty acids, and alpha,omega-diols. The crystal structure of the enzyme bound to octanoic acid provided insights into its regioselectivity and substrate binding orientation.
Article
Chemistry, Multidisciplinary
Thien-Kim Le, Jinhyun Kim, Ngoc Anh Nguyen, Thi Huong Ha Nguyen, Eun-Gene Sun, Su-Min Yee, Hyung-Sik Kang, Soo-Jin Yeom, Chan Beum Park, Chul-Ho Yun
Summary: Photobiocatalysis is a green platform for driving redox enzymatic reactions using solar energy without high-cost cofactors. Natural flavins were used as photosensitizers to develop a visible light-driven whole-cell platform for human cytochrome P450 (CYP) photobiocatalysis. The research showed that flavins can mediate light-driven biocatalysis by human CYPs in whole-cell systems, demonstrating the general applicability of solar-powered P450 photobiocatalytic system.
Article
Chemistry, Multidisciplinary
Sascha Grobe, Christoffel P. S. Badenhorst, Thomas Bayer, Emil Hamnevik, Shuke Wu, Christoph W. Grathwol, Andreas Link, Sven Koban, Henrike Brundiek, Beatrice Grossjohann, Uwe T. Bornscheuer
Summary: In this study, the cytochrome P450 monooxygenase CYP107D1 was engineered for the stereo- and regioselective 7 beta-hydroxylation of lithocholic acid to yield ursodeoxycholic acid. By identifying specificity-determining residues and utilizing a synthetic small library, a nearly perfectly regio- and stereoselective triple mutant was developed, which produces UDCA at significantly higher levels than the initial variant. This biocatalyst offers new possibilities for environmentally friendly synthesis of UDCA from biological waste product.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2021)
Article
Biochemistry & Molecular Biology
Jinia Akter, Tegan P. Stockdale, Stella A. Child, Joel H. Z. Lee, James J. Voss, Stephen G. Bell
Summary: CYP102A1 enzyme can be engineered into a peroxygenase by mutating Thr268Glu, which enhances its hydroxylation activity towards saturated fatty acids and offers the potential for simpler cytochrome P450 systems for selective oxidations. The peroxygenase variant showed similar product distributions as the wild-type enzyme and regioselectively hydroxylated specific positions in the fatty acids tested.
JOURNAL OF INORGANIC BIOCHEMISTRY
(2023)
Article
Biochemistry & Molecular Biology
Freeborn Rwere, Sangchoul Im, Lucy Waskell
Summary: By altering the 140s FMN binding loop of CYPOR, the activity with cyt P450s can be affected, with the mutant activity primarily determined by its ability to reduce ferric cyt P450. The different behaviors of the mutants are mainly due to the altered structure and mobility of the loop.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2021)
Article
Chemistry, Multidisciplinary
Alecia R. Gee, Isobella S. J. Stone, Tegan P. Stockdale, Tara L. Pukala, James J. De Voss, Stephen G. Bell
Summary: We have achieved efficient oxidation reactions on unactivated carbon-hydrogen bonds by modifying the enzyme structure.
CHEMICAL COMMUNICATIONS
(2023)
Review
Chemistry, Applied
Rui Zhu, Yang Liu, Yueying Yang, Qing Min, Hua Li, Lixia Chen
Summary: Steroids are a class of drugs with diverse pharmacological properties. Hydroxylation of steroids significantly affects their biological activities and properties. Achieving regio- and stereo-selective hydroxylation on steroids using modern organic synthesis techniques is extremely challenging. However, recent studies have identified and engineered cytochrome P450 monooxygenases that possess the ability to catalyze regio- and stereo-selective oxidations of nonactivated C-H bonds in complex organic molecules.
ADVANCED SYNTHESIS & CATALYSIS
(2022)
Article
Chemistry, Multidisciplinary
Danilo Correddu, Sabrina Helmy Aly, Giovanna Di Nardo, Gianluca Catucci, Cristina Prandi, Marco Blangetti, Chiara Bellomo, Elisabetta Bonometti, Guido Viscardi, Gianfranco Gilardi
Summary: This study found that the cytochrome P450 BM3 mutants have enhanced and specific epoxidation activity for geraniol and linalool, forming valuable oxidized derivatives, highlighting the significance of these enzymes for industrial applications.
Review
Chemistry, Physical
Donya Valikhani, Juan M. Bolivar, Joelle N. Pelletier
Summary: Cytochrome P450s are a large superfamily of heme proteins with important applications in drug metabolism studies and biocatalytic transformations. Immobilization of P450 enzymes on biosensing platforms and in biocatalytic reactions plays a key role in improving operational stability. Various methods and materials have been developed to address the challenges associated with the unique characteristics of P450s in different applications.
Article
Chemistry, Physical
Chun-Chi Chen, Meng Dai, Lilan Zhang, Jing Zhao, Wei Zeng, Min Shi, Jian-Wen Huang, Weidong Liu, Rey-Ting Guo, Aitao Li
Summary: We investigated the mechanism of the P450tol enzyme from Rhodococcus coprophilus TC-2, which can hydroxylate toluene at the benzylic site. We solved the crystal structures of P450tol and its complex with the substrate, revealing how P450tol restricts the binding position of toluene and enables precise hydroxylation at the benzylic site. Additionally, we found that P450tol can also hydroxylate larger substrates and synthesized a self-sufficient chimeric enzyme with improved activity and stability. These findings highlight the potential of P450tol in biotechnological and pharmaceutical applications.
Review
Biotechnology & Applied Microbiology
Danilo Correddu, Giovanna Di Nardo, Gianfranco Gilardi
Summary: Members of class VII cytochromes P450 are catalytically self-sufficient enzymes that have been isolated from extremophiles or detoxifying bacteria. Protein engineering has generated mutants capable of performing specific organic reactions, which can be further researched for biotechnological applications. The detailed 3D structure of these enzymes provides a basis for directed evolution studies aimed at expanding their biotechnological potential.
TRENDS IN BIOTECHNOLOGY
(2021)
Article
Chemistry, Physical
Keita Omura, Yuichiro Aiba, Kazuto Suzuki, Shinya Ariyasu, Hiroshi Sugimoto, Osami Shoji
Summary: The self-sufficient enzyme P450BM3 was reconstituted with manganese protoporphyrin IX, resulting in Mn-BM3. This artificial enzyme was found to catalyze the monooxygenation of various substrates by activating molecular oxygen. The study also revealed the involvement of a radical intermediate in the hydroxylation process.
Review
Chemistry, Multidisciplinary
Frank Hollmann, Diederik J. Opperman, Caroline E. Paul
Summary: This review highlights the synthetic state-of-the-art and potential of enzymes catalyzing reductions, ranging from carbonyl, enone, and aromatic reductions to reductive aminations. Enzymes play a key role in producing chiral products with new functionalities, leading to alternative synthetic routes for high-added-value compounds and bulk chemicals.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2021)
Article
Chemistry, Physical
Nathalie Nett, Sabine Duewel, Luca Schmermund, Gerrit E. Benary, Kara Ranaghan, Adrian Mulholland, Diederik J. Opperman, Sabrina Hoebenreich
Summary: The engineered panel of ene-reductases (ERs) from Thermus scotoductus SA-01 shows control over facial selectivity, thermostability, organic solvent tolerance, and a broad substrate scope. The TsER variant panel exhibits excellent enantiomeric excess (ee) and yields during bi-phasic preparative scale synthesis.
MOLECULAR CATALYSIS
(2021)
Article
Multidisciplinary Sciences
Minkyung Baek, Frank DiMaio, Ivan Anishchenko, Justas Dauparas, Sergey Ovchinnikov, Gyu Rie Lee, Jue Wang, Qian Cong, Lisa N. Kinch, R. Dustin Schaeffer, Claudia Millan, Hahnbeom Park, Carson Adams, Caleb R. Glassman, Andy DeGiovanni, Jose H. Pereira, Andria V. Rodrigues, Alberdina A. van Dijk, Ana C. Ebrecht, Diederik J. Opperman, Theo Sagmeister, Christoph Buhlheller, Tea Pavkov-Keller, Manoj K. Rathinaswamy, Udit Dalwadi, Calvin K. Yip, John E. Burke, K. Christopher Garcia, Nick V. Grishin, Paul D. Adams, Randy J. Read, David Baker
Summary: Through the three-track network, we achieved accuracies approaching those of DeepMind in CASP14, enabling rapid solution of challenging x-ray crystallography and cryo-electron microscopy structure modeling problems, and providing insights into the functions of proteins with currently unknown structure.
Article
Biochemistry & Molecular Biology
Cintia Soledad Ramirez, Carmien Tolmie, Diederik Johannes Opperman, Pablo Javier Gonzalez, Maria Gabriela Rivas, Carlos Dante Brondino, Felix Martin Ferroni
Summary: This study presents the crystal structure of a copper-containing nitrite reductase (NirK) from Sm and its role in symbiosis, as well as complex structural alignments and docking studies with related proteins. The activity of SmNirK, although lower than classical NirKs, is affected by structural differences and regulation within the symbiotic environment. In vitro kinetics demonstrated similar electron donation performance from SmPaz1 and SmPaz2 to SmNirK, facilitated by conserved residues mediating interprotein electron transfer.
Article
Chemistry, Physical
Cheri Louise Jacobs, Rodolpho do Aido-Machado, Carmien Tolmie, Martha Sophia Smit, Diederik Johannes Opperman
Summary: Selective oxyfunctionalization of non-activated C-H bonds is a significant challenge in synthetic chemistry. In this study, the biocatalytic hydroxylation of medium-chain n-alkanes by the bacterial enzyme CYP153A71 was characterized. The enzyme was found to convert 1-alkanols to aldehydes, fatty acids, and alpha,omega-diols. The crystal structure of the enzyme bound to octanoic acid provided insights into its regioselectivity and substrate binding orientation.
Article
Biotechnology & Applied Microbiology
Martha Sophia Smit, Mpeyake Jacob Maseme, Jacqueline van Marwijk, Jasmin Cara Aschenbrenner, Diederik Johannes Opperman
Summary: Cytochrome P450 monooxygenases (CYP450s) are crucial in the synthesis and breakdown of secondary metabolites in eukaryotes, especially in plants and fungi. CYP505E3 from Aspergillus terreus demonstrates remarkable omega-7 hydroxylase activity, making it a promising catalyst for delta-dodecalactone synthesis. Other CYP505Es also display similar activity, but their occurrence is limited to certain species like Aspergillus and Penicillium.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2023)
Article
Chemistry, Multidisciplinary
Gabriela C. Schroeder, Martha S. Smit, Diederik J. Opperman
Summary: CYPs/P450s are versatile hemethiolate proteins with high stereo- and regio-selectivity, making them powerful biocatalysts for synthesizing value-added compounds. This review highlights recent advances in P450-mediated syntheses, including enzyme engineering strategies to enhance selectivity and facilitate bond formation, as well as approaches to overcome limitations of P450 catalysis for improved efficiency and biotechnological viability.
CURRENT OPINION IN GREEN AND SUSTAINABLE CHEMISTRY
(2023)
Article
Chemistry, Organic
Ana C. Ebrecht, Thato M. Mofokeng, Frank Hollmann, Martha S. Smit, Diederik J. Opperman
Summary: γ- and δ-lactones are important flavor and fragrance compounds, and their synthesis relies on suitable hydroxy fatty acid precursors. Three short unspecific peroxygenases were found to selectively hydroxylate C4 and C5 positions of C8-C12 fatty acids, leading to the corresponding γ- and δ-lactones after lactonization. The preference for C4 hydroxylation resulted in γ-lactones as the major products. Overoxidation of the hydroxy fatty acids was addressed by reducing the resulting oxo acids using an alcohol dehydrogenase in a bienzymatic cascade reaction.
Article
Chemistry, Physical
Jasmin C. Aschenbrenner, Ana C. Ebrecht, Carmien Tolmie, Martha S. Smit, Diederik J. Opperman
Summary: CYP505A30 is a fungal cytochrome P450 monooxygenase that can selectively oxidize n-alkanes, fatty alcohols, and fatty acids. Structural analysis reveals high similarity with bacterial CYP102A1, but differences in the active site suggest potential for rational mutation to alter regioselectivity. Mutations introduced increased regioselectivity for specific n-alkanes, providing a pathway for more selective production of non-vicinal diols.
CATALYSIS SCIENCE & TECHNOLOGY
(2021)
Article
Chemistry, Multidisciplinary
Derik Wilbers, Joseph Brehm, Richard J. Lewis, Jacqueline van Marwijk, Thomas E. Davies, David J. Morgan, Diederik J. Opperman, Martha S. Smit, Miguel Alcalde, Athanasios Kotsiopoulos, Susan T. L. Harrison, Graham J. Hutchings, Simon J. Freakley
Summary: The combination of heterogeneous catalysis and biocatalysis to form one-pot reaction cascades allows the integration of enzymatic transformations into existing chemical infrastructure. The use of peroxygenases for clean C-H activation in such tandem systems requires a constant supply of low-level hydrogen peroxide. The in situ generation of oxidants from cheap reactants at industrial scale shows potential for highly active cascade processes.
Article
Chemistry, Organic
Ana C. Ebrecht, Jasmin C. Aschenbrenner, Martha S. Smit, Diederik J. Opperman
Summary: CYP505A30 shows substrate promiscuity by not only regioselectively oxyfunctionalizing fatty acids and primary fatty alcohols, but also readily accepting alkanes for the production of non-vicinal diols through sequential oxygenation. This biocatalyst with versatile substrate specificity and activity has promising applications in various fields.
ORGANIC & BIOMOLECULAR CHEMISTRY
(2021)