Article
Pharmacology & Pharmacy
Songya Zhang, Lin Zhang, Anja Greule, Julien Tailhades, Edward Marschall, Panward Prasongpholchai, Daniel J. Leng, Jingfan Zhang, Jing Zhu, Joe A. Kaczmarski, Ralf B. Schittenhelm, Oliver Einsle, Colin J. Jackson, Fabrizio Alberti, Andreas Bechthold, Youming Zhang, Manuela Tosin, Tong Si, Max J. Cryle
Summary: WS9326A is a peptide antibiotic synthesized by a non-ribosomal peptide synthetase (NRPS). The cytochrome P450 encoded by sas16 (P450Sas) is critical for the formation of an unusual amino acid residue in WS9326A. In this study, the researchers identified the substrate of P450Sas and elucidated its role in the biosynthetic pathway of WS9326A. The results suggest that P450Sas catalyzes the direct dehydrogenation of a dipeptide intermediate, expanding the range of P450 enzymes that can be used for the production of biologically active peptides.
ACTA PHARMACEUTICA SINICA B
(2023)
Article
Chemistry, Multidisciplinary
Anja Greule, Thierry Izore, Daniel Machell, Mathias H. Hansen, Melanie Schoppet, James J. De Voss, Louise K. Charkoudian, Ralf B. Schittenhelm, Jeffrey R. Harmer, Max J. Cryle
Summary: Cytochrome P450 enzymes are enzymes that can perform oxidative reactions using heme. However, some of these enzymes can be damaged by oxidation, leading to decreased enzyme activity. Research has shown that the heme orientation in the active site of certain cytochrome P450 enzymes may contribute to this oxidative damage.
FRONTIERS IN CHEMISTRY
(2022)
Article
Chemistry, Multidisciplinary
Hulya Aldemir, Shuangjie Shu, Francoise Schaefers, Hanna Hong, Rene Richarz, Sabrina Harteis, Manuel Einsiedler, Tobias M. Milzarek, Sabine Schneider, Tobias A. M. Gulder
Summary: The arylomycin antibiotics are potent inhibitors of bacterial type I signal peptidase, containing a biaryl structural motif reminiscent of glycopeptide antibiotics. AryC, a cytochrome P450 enzyme, performs biaryl coupling in arylomycin biosynthesis without the need for any protein interaction partner, due to its strongly hydrophobic cavity at the surface pointing to the substrate tunnel. This unique reactivity enables chemo-enzymatic assembly of arylomycin A2 combining liquid- and solid-phase peptide synthesis advantages.
CHEMISTRY-A EUROPEAN JOURNAL
(2022)
Article
Biochemistry & Molecular Biology
Lara E. Zetzsche, Suman Chakrabarty, Alison R. H. Narayan
Summary: This study optimized a whole-cell biocatalytic platform for the synthesis of biaryl molecules in yeast through recombinant production of the fungal P450 KtnC. Moreover, engineering redox self-sufficient fusion enzymes further enhanced the efficiency of the system. This provides a platform for engineering currently underexplored fungal P450s into selective biocatalysts for the synthesis of complex biaryl compounds.
ACS CHEMICAL BIOLOGY
(2022)
Article
Biochemistry & Molecular Biology
Joshua Kyle Stanfield, Hiroki Onoda, Shinya Ariyasu, Chie Kasai, Eleanor Mary Burfoot, Hiroshi Sugimoto, Osami Shoji
Summary: Cytochrome P450 enzymes have shown promise as biocatalysts in environmentally friendly catalytic systems. Here, we demonstrate that decoy molecules can trick CYP102A5 and A7 into becoming active and hydroxylating non-native substrates. Additionally, differences in decoy molecule selectivity and binding were observed.
JOURNAL OF INORGANIC BIOCHEMISTRY
(2023)
Article
Chemistry, Multidisciplinary
Yuanyuan Shi, Zhibo Jiang, Xiaowen Hu, Xiaomin Hu, Renjie Gu, Bingya Jiang, Lijie Zuo, Xingxing Li, Hongmin Sun, Cong Zhang, Lifei Wang, Linzhuan Wu, Bin Hong
Summary: The study elucidated the C-S bond forming activity of cytochrome P450 monooxygenase CxnD in chuangxinmycin biosynthesis, showing that CxnD can generate a unique skeleton structure through in vivo and in vitro analyses. The X-ray crystal structure and structure-based mutagenesis revealed details of substrate binding mode.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2021)
Review
Biochemistry & Molecular Biology
Wolfgang Huettel, Michael Mueller
Summary: Phenol coupling reaction is crucial in the biosynthesis, but challenging to control selectivity. Various enzymes catalyze this reaction, especially laccases with complex catalytic systems affecting activity and selectivity. Although the field of enzymatic phenol coupling is still in its infancy, the recent identification of diverse enzymes could facilitate biotechnological development.
NATURAL PRODUCT REPORTS
(2021)
Article
Biochemistry & Molecular Biology
Nicole Hauser, Kendra A. Ireland, Vasiliki T. Chioti, Clarissa C. Forneris, Katherine M. Davis, Mohammad R. Seyedsayamdost
Summary: The emergence of multidrug-resistant pathogens poses a threat to public health, calling for new antimicrobial agents. Vancomycin, as the archetypal glycopeptide antibiotic used against drug-resistant Gram-positive pathogens, provides a promising starting point. The recent chemoenzymatic synthesis of vancomycin suggests that this approach can be widely applied.
ACS CHEMICAL BIOLOGY
(2023)
Article
Biochemistry & Molecular Biology
Yvonne Carius, Michael Hutter, Flora Kiss, Rita Bernhardt, C. Roy D. Lancaster
Summary: This study revealed the crystal structure of cytochrome P450 CYP106A1 from Priestia megaterium, which shows a rare sixth beta-sheet. Comparison with the previously studied CYP106A2 and docking studies with the substrate cortisol revealed a different orientation of the steroid molecule in the active sites, explaining the observed differences in substrate conversion and product formation by the two enzymes.
Article
Biochemistry & Molecular Biology
Irina F. Sevrioukova
Summary: CYP3A7 is a fetal/neonatal liver enzyme involved in estriol synthesis and xenobiotic metabolism, with reduced ligand binding capacity compared to adult liver enzyme CYP3A4. Structural analysis of CYP3A7 reveals decreased flexibility and altered interactions in the active site, limiting protein plasticity and potentially impacting reactivity and oxidation of substrates.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2021)
Article
Biochemistry & Molecular Biology
Pradeep Subedi, Hackwon Do, Jun Hyuck Lee, Tae-Jin Oh
Summary: Cytochrome P450 enzymes are heme-containing enzymes that catalyze hydroxylation of various biological molecules, with substrates selectivity possibly governed by highly flexible loops and unique sequences around the substrate entrance region. The newly identified CYP101D5 from Sphingomonas echinoides catalyzes hydroxylation of beta-ionone and flavonoids, suggesting that spatial constraints at the substrate-recognition site originate from the B/C loop. Charge distribution at the substrate binding site may also play a role in substrate selectivity and preference for CYP101D5.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Article
Biochemistry & Molecular Biology
Christopher S. Campomizzi, George E. Ghanatios, D. Fernando Estrada
Summary: Cytochromes P450 are versatile enzymes involved in endogenous and exogenous metabolism, undergoing structural changes related to function. This study demonstrates the utility of fluorine (19F)-NMR spectroscopy in monitoring structural changes in CYP121A1, revealing insights into its role in substrate recognition and mechanistic details of this essential enzyme from Mycobacterium tuberculosis.
JOURNAL OF BIOLOGICAL CHEMISTRY
(2021)
Article
Biochemistry & Molecular Biology
Chao Shi, Mingyue Gu, Zhangxin Chen, Xiaonan Huang, Juan Guo, Luqi Huang, Jie Deng, Ke He, Lei Zhang, Lixin Huang, Zhenzhan Chang
Summary: The study revealed the crystal structures of CYP76AH1 and its natural substrate miltiradiene, providing important insights into its catalytic mechanism and the basis for improving tanshinone production.
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
(2021)
Article
Chemistry, Multidisciplinary
Yongwei Zhao, Edward Marschall, Maxine Treisman, Alasdair McKay, Leo Padva, Max Crusemann, David R. Nelson, David L. Steer, Ralf B. Schittenhelm, Julien Tailhades, Max J. Cryle
Summary: This study investigates the use of peptide crosslinking cytochrome P450 enzymes in generating cyclic tripeptides from simple synthons. The enzymes produced both tyrosine-histidine and tyrosine-tryptophan crosslinked tripeptides, the latter being a rare example of phenolic crosslinking to an indole moiety. The tripeptides can be easily isolated after removing the leader peptide and can include a wide range of amino acids. This research suggests that P450 enzymes have the potential to play a significant role in the synthesis of high-value cyclic tripeptides, which can be further diversified using selective chemical techniques.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2022)
Article
Biotechnology & Applied Microbiology
Sheng Dong, Jingfei Chen, Xingwang Zhang, Fei Guo, Li Ma, Cai You, Xiao Wang, Wei Zhang, Xiaobo Wan, Shuang-Jiang Liu, Li-Shan Yao, Shengying Li, Lei Du, Yingang Feng
Summary: Selective oxidation of C-H bonds in alkylphenols is crucial for structural derivatization in pharmaceutical and biomanufacturing, as well as for biological degradation of toxic chemicals in environmental protection. A novel chemomimetic biocatalytic system using cytochrome P450 monooxygenase CreJ has been developed, showing excellent regio- and stereoselectivity in oxidizing various alkylphenol substrates. The results provide mechanistic insights into CreJ regio- and stereoselectivity, offering promising directions for further enzyme design and engineering efforts.
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
(2021)
Article
Multidisciplinary Sciences
Thierry Izore, Julien Tailhades, Mathias Henning Hansen, Joe A. Kaczmarski, Colin J. Jackson, Max J. Cryle
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
(2019)
Article
Biochemistry & Molecular Biology
Terry C. C. Lim Kam Sian, Saranjah Indumathy, Hanim Halim, Anja Greule, Max J. Cryle, Paul Bowness, Jamie Rossjohn, Stephanie Gras, Anthony W. Purcell, Ralf B. Schittenhelm
JOURNAL OF BIOLOGICAL CHEMISTRY
(2019)
Article
Chemistry, Organic
Yongwei Zhao, Robert J. A. Goode, Ralf B. Schittenhelm, Julien Tailhades, Max J. Cryle
JOURNAL OF ORGANIC CHEMISTRY
(2020)
Article
Biochemistry & Molecular Biology
Milda Kaniusaite, Julien Tailhades, Tiia Kittila, Christopher D. Fage, Robert J. A. Goode, Ralf B. Schittenhelm, Max J. Cryle
Summary: The biosynthesis of glycopeptide antibiotics demonstrates the exceptional ability of nonribosomal peptide synthesis to generate diverse and complex structures. An important study on the NRPS assembly lines involved in synthesizing GPAs revealed that they function as dynamic peptide assembly lines, allowing for flexible control over amino acid modifications and peptide formation, which is beneficial for the redesign of important biosynthetic systems.
Article
Microbiology
Rhys Grinter, Blair Ney, Rajini Brammananth, Christopher K. Barlow, Paul R. F. Cordero, David L. Gillett, Thierry Izore, Max J. Cryle, Liam K. Harold, Gregory M. Cook, George Taiaroa, Deborah A. Williamson, Andrew C. Warden, John G. Oakeshott, Matthew C. Taylor, Paul K. Crellin, Colin J. Jackson, Ralf B. Schittenhelm, Ross L. Coppel, Chris Greening
Review
Biochemistry & Molecular Biology
Yongwei Zhao, Y. T. Candace Ho, Julien Tailhades, Max Cryle
Summary: Researchers are intrigued by the complex biosynthesis of glycopeptide antibiotics, particularly the role of the X-domain in recruiting P450 enzymes. In vitro studies have provided insights into the tolerances and limitations of the GPA cyclisation cascade, paving the way for future reengineering of this important antibiotic class.
Article
Biochemistry & Molecular Biology
Milda Kaniusaite, Tiia Kittila, Robert J. A. Goode, Ralf B. Schittenhelm, Max J. Cryle
ACS CHEMICAL BIOLOGY
(2020)
Letter
Biochemistry & Molecular Biology
Daniel L. Machell, Mathias H. Hansen, Max J. Cryle
Summary: Detection of pyrophosphate is crucial for quantifying enzyme activity, especially adenylation domain activity in non-ribosomal peptide synthesis. After screening, viable replacement enzymes with higher activity have been identified, allowing the continued use of the established online assay for pyrophosphate detection.
Article
Biochemistry & Molecular Biology
Y. T. Candace Ho, Ralf B. Schittenhelm, Dumitrita Iftime, Evi Stegmann, Julien Tailhades, Max J. Cryle
Summary: Glycopeptide antibiotics inhibit bacterial cell-wall biosynthesis by sequestration of precursor lipid II. The oxidative crosslinking of the core peptide during GPA biosynthesis is essential and challenging. Understanding the activity and selectivity of Oxy enzymes is important for future engineering of this compound class.
Article
Chemistry, Multidisciplinary
Rania A. A. Hashad, Edwina Jap, Joanne L. L. Casey, Y. T. Candace Ho, Alexander Wright, Claudia Thalmann, Mark Sleeman, David W. W. Lupton, Christoph E. E. Hagemeyer, Max J. J. Cryle, Remy Robert, Karen Alt
Summary: A highly effective 2-step system using engineered methionine residues was used for site-specific antibody modification and conjugation. This system offers a novel way to fundamentally change the process of antibody bioconjugation. The versatility of this system was demonstrated by incorporating a fluorescent dye and can be applied to a wide variety of conjugation partners.
CHEMISTRY-A EUROPEAN JOURNAL
(2023)
Review
Biochemistry & Molecular Biology
Songya Zhang, Yunliang Chen, Jing Zhu, Qiujie Lu, Max J. Cryle, Youming Zhang, Fu Yan
Summary: Streptomyces bacteria are widely distributed in terrestrial and marine environments and are a rich source of active natural products due to their metabolic diversity. This review highlights the importance of nonribosomal lipopeptides as important natural products with diverse biological activities that play crucial roles in the lifestyle of Streptomyces. Recent advances in the biosynthesis of lipopeptide antibiotics produced by Streptomyces have greatly contributed to our understanding of their structures, properties, biosynthetic mechanisms, chemical and chemoenzymatic synthesis, and biological functions. The use of genome mining techniques has led to the discovery of many novel lipopeptides, further demonstrating their potential for future development in modern medicine.
NATURAL PRODUCT REPORTS
(2023)
Article
Chemistry, Multidisciplinary
Y. T. Candace Ho, Joe A. Kaczmarski, Julien Tailhades, Thierry Izore, David L. Steer, Ralf B. Schittenhelm, Manuela Tosin, Colin J. Jackson, Max J. Cryle
Summary: Nonribosomal peptide synthetases play a significant role in producing essential peptide natural products, with carrier proteins as their core component. By replacing the CP substrate thioesters with stabilized ester analogues, active condensation domain complexes are formed, while amide stabilization leads to non-functional complexes.
CHEMICAL COMMUNICATIONS
(2023)
Article
Chemistry, Multidisciplinary
Jennifer A. E. Payne, Ketav Kulkarni, Thierry Izore, Alex J. Fulcher, Anton Y. Peleg, Marie-Isabel Aguilar, Max J. Cryle, Mark P. Del Borgo
Summary: A new antimicrobial material was designed, synthesised and characterised by the self-assembly of two distinct beta-peptide monomers, which showed inhibitory effects on drug-resistant bacteria like MRSA. This study provides insights into the design of peptide-based supramolecular assemblies with antibacterial activity.
NANOSCALE ADVANCES
(2021)
Article
Chemistry, Multidisciplinary
Daniel J. Leng, Anja Greule, Max J. Cryle, Manuela Tosin
Summary: Chemical probes were used to capture biosynthetic intermediates generated in the nonribosomal peptide formation of vancomycin in vivo. The putative intercepted intermediates were characterised via HR-LC-MS2, providing insights into the timing of the first chlorination of the peptide backbone by the halogenase VhaA. This information is significant for enzyme engineering towards the production of novel glycopeptides.
CHEMICAL COMMUNICATIONS
(2021)
Article
Chemistry, Multidisciplinary
Milda Kaniusaite, Robert J. A. Goode, Julien Tailhades, Ralf B. Schittenhelm, Max J. Cryle