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)
Article
Biotechnology & Applied Microbiology
Ming Yang, Wanlu Li, Lin Zhou, Xiao Lin, Wenyu Zhang, Yaoyao Shen, Hai Deng, Hou-wen Lin, Yongjun Zhou
Summary: This article investigates the synthesis of bacterial trialkyl-substituted aromatic polyketides with antidiabetic and immunosuppressant activities. Through gene deletion and complementation experiments, two essential genes for the biosynthesis of these compounds were identified and their functions were analyzed.
SYNTHETIC AND SYSTEMS BIOTECHNOLOGY
(2023)
Article
Chemistry, Organic
Jordan T. Brazeau-Henrie, Andre R. Paquette, Allison Q. . O'Rourke, Michael G. Darnowski, Christopher N. Boddy
Summary: The total and chemoenzymatic synthesis of the depsipeptide natural product seongsanamide E, 3, is described. The synthetic C-terminal N-acetylcysteamine thioester of linear natural product 1 was macrolactonized by the excised recombinant purified seongsanamide thioesterase (Sgd-TE) domain, generating 3. Sgd-TE also effects the ring opening of 3. Chemical synthesis provided 3 through a macrolactamization strategy. This work confirms the biosynthesis of 3 and demonstrates the power of Sgd-TE as a biocatalyst.
Article
Multidisciplinary Sciences
Woo Cheol Lee, Sungjae Choi, Ahjin Jang, Kkabi Son, Yangmee Kim
Summary: The study focused on the crystal structures of the beta-ketoacyl-ACP reductase ApeQ produced by an APE gene cluster in Gram-negative bacteria. It was found that ApeQ plays a crucial role in APE biosynthesis and cannot be substituted by a similar enzyme from the FAS gene cluster. Structural and functional analysis revealed a positively charged arginine patch in ApeQ as the cognate ACP-binding site.
SCIENTIFIC REPORTS
(2021)
Article
Biochemistry & Molecular Biology
Waraporn Bunnak, Ashley J. Winter, Colin M. Lazarus, Matthew P. Crump, Paul R. Race, Pakorn Wattana-Amorn
Summary: This study reports the solution characterization of singlet and doublet acyl carrier protein (ACP(2) and ACP(1)-ACP(2))-thioesterase (TE) domains from NR-PKS involved in menisporopsin A biosynthesis. Small-angle X-ray scattering (SAXS) studies in combination with homology modelling reveal that these polypeptides adopt a distinctive beads-on-a-string configuration, characterized by highly flexible interdomain linkers. These models provide a platform for studying domain organization and interdomain interactions in fungal NR-PKSs, which may aid in designing functionally optimized polyketide scaffolds.
Article
Biochemistry & Molecular Biology
Peter D. Giang, Luke R. Churchman, Jeanette E. Stok, Rochelle M. Soo, James J. De Voss
Summary: A study found that Rhodococcus globerulus (R. globerulus) isolated from soil beneath Eucalyptus sp. can thrive on mono-terpenes such as 1,8-cineole, p-cymene and (R)- and (S)-limonene as its sole sources of carbon and energy. The research revealed a novel cytochrome P450 (CYP108N12) in R. globerulus which may be responsible for initiating the biodegradation of structurally similar monoterpene compounds. This finding suggests that CYP108N12 has potential as a biocatalyst with specific hydroxylation capabilities.
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS
(2022)
Article
Chemistry, Physical
Zhan Song, Meijing Wei, Yinghao Fang, Fuping Lu, Minze Jia, Hui-Min Qin, Shuhong Mao
Summary: Custom exogenous ferredoxin (Fdx) was employed to enhance the conversion rate of 4-androstene-3,17-dione (AD) by facilitating electron transfer between reductase and oxygenase through a newly created conduit. The specific amino acid residues in the multienzyme complex responsible for direct electron transfer (DET) pathways were analyzed and some unique residues from the reductase or ferredoxin domains were identified. The electrostatic interactions in mutant MT2 (Y110R in KshA, T85D in DmFdx2) were strengthened using an iterative saturation mutagenesis (ISM) strategy. Moreover, Fdx was proposed as a scaffold for immobilization and self-assembly of the multi-enzyme complex, leading to increased AD conversion rates.
CATALYSIS SCIENCE & TECHNOLOGY
(2023)
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
Biotechnology & Applied Microbiology
Ling Wang, Xiaowei Wu, Chenghua Gao, Lingrui Wei, Qian Li, Aitao Li
Summary: A new cytochrome P450 enzyme, CYP68J5_fg, was identified from Fusarium graminearum, showing steroidal 12 beta- and 15 alpha-hydroxylase activities. This is the first fungal P450 enzyme with 12 beta-hydroxylase activity discovered, offering opportunities for enhancing efficiency and selectivity in steroid drug synthesis.
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
(2023)
Article
Biochemistry & Molecular Biology
Yen-Ting Lin, Sam P. de Visser
Summary: There are two types of cytochrome P450 enzymes in nature: monooxygenases and peroxygenases. They play roles in substrate degradation or biosynthesis, with monooxygenases using dioxygen and peroxygenases using H2O2 in their catalytic cycles. P450 peroxygenases do not require external redox partners or proton sources, making them self-sufficient and promising for biotechnological applications.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2021)
Article
Chemistry, Multidisciplinary
Jie Chen, Sheng Dong, Wenhan Fang, Yiping Jiang, Zhifeng Chen, Xiangquan Qin, Cong Wang, Haifeng Zhou, Longyi Jin, Yingang Feng, Binju Wang, Zhiqi Cong
Summary: We report the selective hydroxylation of alkylbenzenes using an engineered P450 peroxygenase driven by a dual-functional small molecule (DFSM). By combining different P450BM3 variants with DFSMs, more than half of all possible hydroxylated products were obtained from each substrate with high regioselectivity (>99%), enantioselectivity (>99% ee), and high total turnover numbers (up to 80963). Synergistic effects between exogenous DFSMs and the protein environment controlled the regio- and enantioselectivity, as revealed by crystal structure analysis, molecular dynamic simulations, and theoretical calculations. This work has implications for exogenous-molecule-modulated enzymatic regiodivergent and enantioselective hydroxylation with potential applications in synthetic chemistry.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2023)
Review
Chemistry, Multidisciplinary
Lynn Buyachuihan, Franziska Stegemann, Martin Grininger
Summary: Megasynthases are multienzyme complexes responsible for producing primary metabolites and complex natural products. They build fatty acids and polyketides stepwise by assembling and modifying small acyl moieties. The shuttling of growing intermediates between catalytic domains is mediated by acyl carrier proteins (ACPs). This review presents the current understanding of substrate shuttling and proposes a mechanism for programming ACP movement in megasynthases.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2023)
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
Xiaodong Zhang, Yun Hu, Wei Peng, Chenghua Gao, Qiong Xing, Binju Wang, Aitao Li
Summary: In this study, the oxidizing activity of CYP109B1 was reconstituted by coupling redox pairs or fusing it to the reductase domain of two self-sufficient P450 enzymes. The best redox pair for CYP109B1 was found to be Fdr_0978/Fdx_1499 from Synechococcus elongatus, showing over 99% conversion with 73% 15 beta selectivity for testosterone. The enzyme displayed good activity and selectivity toward testosterone derivatives, with a shift in selectivity for specific substrates.
FRONTIERS IN CHEMISTRY
(2021)
Article
Chemistry, Physical
Shinya Ariyasu, Kai Yonemura, Chie Kasai, Yuichiro Aiba, Hiroki Onoda, Yuma Shisaka, Hiroshi Sugimoto, Takehiko Tosha, Minoru Kubo, Takashi Kamachi, Kazunari Yoshizawa, Osami Shoji
Summary: This study reports the catalytic oxidation of methane to methanol by wild-type P450BM3 without mutagenesis. The findings have significant implications for the development of biological methane oxidation technology.
Article
Genetics & Heredity
Cerith Jones, Gordon Webster, Alex J. Mullins, Matthew Jenner, Matthew J. Bull, Yousef Dashti, Theodore Spilker, Julian Parkhill, Thomas R. Connor, John J. LiPuma, Gregory L. Challis, Eshwar Mahenthiralingam
Summary: Burkholderia gladioli is a diverse bacterium with significant ecological impacts on humans, animals, and plants, spanning diseases and beneficial interactions. Through exploring its genomic diversity and metabolic potential, it has shed light on its roles as a plant pathogen, food poisoning agent, and lung pathogen for CF patients. This research has identified potential therapeutic strategies to minimize poisoning risks associated with this bacterium.
MICROBIAL GENOMICS
(2021)
Article
Microbiology
Matthieu Nicault, Ali Zaiter, Stephane Dumarcay, Patrick Chaimbault, Eric Gelhaye, Pierre Leblond, Cyril Bontemps
Summary: The study explored the impact of co-culture between Streptomyces and fungi on the production of bioactive compounds, finding that co-culture had a significant effect on the metabolic expression of each partner and facilitated the expression of specific compounds.
Article
Ecology
Valentin Waschulin, Chiara Borsetto, Robert James, Kevin K. Newsham, Stefano Donadio, Christophe Corre, Elizabeth Wellington
Summary: The study used long-read sequencing and genome mining to recover over 1400 mostly full-length biosynthetic gene clusters (BGC) from soil in Mars Oasis, Antarctica, demonstrating the rich diversity of BGCs from uncultivated bacterial lineages. Highly divergent BGCs were found in various phyla including a potential novel family of RiPPs, highlighting the biosynthetic potential of underexplored phyla and unexplored lineages within well-studied producer phyla. The findings showcase long-read metagenomic sequencing as a promising approach to accessing the genetic reservoir of specialized metabolite gene clusters in the uncultured majority of microbes.
Article
Multidisciplinary Sciences
Virginia S. Lioy, Jean-Noel Lorenzi, Soumaya Najah, Thibault Poinsignon, Herve Leh, Corinne Saulnier, Bertrand Aigle, Sylvie Lautru, Annabelle Thibessard, Olivier Lespinet, Pierre Leblond, Yan Jaszczyszyn, Kevin Gorrichon, Nelle Varoquaux, Ivan Junier, Frederic Boccard, Jean-Luc Pernodet, Stephanie Bury-Mone
Summary: The linear chromosome structure of Streptomyces bacteria is linked to genetic compartmentalization, and the onset of metabolic differentiation involves rearrangement of chromosome architecture.
NATURE COMMUNICATIONS
(2021)
Article
Biochemistry & Molecular Biology
Katarzyna H. Maslowska, Florencia Villafanez, Luisa Laureti, Shigenori Iwai, Vincent Pages
Summary: This study reveals that Translesion Synthesis (TLS) is locally controlled by PCNA ubiquitination at each individual lesion after a genotoxic stress. Activation of the DDR does not promote TLS and mutagenesis, in contrast to the SOS response in bacteria.
NUCLEIC ACIDS RESEARCH
(2022)
Article
Biochemistry & Molecular Biology
Jean-Hugues Guervilly, Marion Blin, Luisa Laureti, Emilie Baudelet, Stephane Audebert, Pierre-Henri Gaillard
Summary: The tumour suppressor SLX4 interacts with MSH2 and regulates the activity of MutS beta and MutS alpha, uncovering an unexpected role of SLX4 in inhibiting repair activity of MMR.
NUCLEIC ACIDS RESEARCH
(2022)
Article
Multidisciplinary Sciences
Li Su, Laurence Hotel, Cedric Paris, Clara Chepkirui, Alexander O. Brachmann, Joern Piel, Christophe Jacob, Bertrand Aigle, Kira J. Weissman
Summary: In this study, the authors reprogrammed the stambomycin PKS by deleting internal modules, resulting in the production of 37-membered mini-stambomycin metabolites. However, the yield of the mini-stambomycin products was reduced and substantial amounts of shunt metabolites were also produced. The findings also revealed an unprecedented off-loading mechanism of stalled intermediates. These discoveries contribute to the understanding of factors that affect the productivity of engineered assembly lines.
NATURE COMMUNICATIONS
(2022)
Article
Chemistry, Organic
Yongchen Wang, Venkaiah Chintalapudi, Haraldur G. Gudmundsson, Gregory L. Challis, Edward A. Anderson
Summary: The stereochemical assignment of macrolide antibiotics stambomycins A-D was based on sequence analysis of the associated polyketide synthase, except for two stereocentres at C28 and C50. The synthesis of two C50 diastereomers of the C33-C51 region of the stambomycins confirms the PKS-based configurational assignment and establishes a strategy for accessing the extended stambomycin framework.
ORGANIC CHEMISTRY FRONTIERS
(2022)
Article
Biochemical Research Methods
Laurence Legon, Christophe Corre, Declan G. Bates, Ahmad A. Mannan
Summary: gcFront is a software tool that uses genetic algorithms to quickly find optimal gene knockout designs and coupling strength for synthesis. It provides users with flexibility in selecting experimental designs.
Article
Biochemistry & Molecular Biology
Abigail L. Whittock, Jack M. Woolley, Nazia Auckloo, Christophe Corre, Vasilios G. Stavros
Summary: This study investigates the relaxation pathway and long-term photostability of mycosporine-like amino acids. The results reveal that the excited state lifetimes of these molecules are short and energy is transferred to the solvent through a conical intersection. High levels of photostability are observed under continuous irradiation.
Article
Genetics & Heredity
Luisa Laureti, Lara Lee, Gaelle Philippin, Michel Kahi, Vincent Pages
Summary: In this study, the researchers found that the presynaptic phase is crucial for modulating lesion tolerance pathways. Inhibition of the extension of the ssDNA gap or the loading of RecA leads to a decrease in Homology Directed Gap Repair (HDGR) and an increase in Translesion Synthesis (TLS). On the other hand, defects in the postsynaptic phase do not increase TLS. The authors suggest that defects in the presynaptic phase delay the formation of the D-loop and increase the time window allowed for TLS.
Article
Biochemistry & Molecular Biology
Christian Hobson, Matthew Jenner, Xinyun Jian, Daniel Griffiths, Douglas M. Roberts, Matias Rey-Carrizo, Gregory L. Challis
Summary: This study reports the discovery of a new variant of a dehydratase domain that eliminates two molecules of water in sequential steps during polyketide chain assembly, leading to the introduction of a conjugated E,Z-diene. These findings expand the catalytic versatility of modular PKSs and highlight the role of a conserved histidine residue in the reaction.
NATURE CHEMICAL BIOLOGY
(2022)
Correction
Multidisciplinary Sciences
Li Su, Laurence Hotel, Cedric Paris, Clara Chepkirui, Alexander O. Brachmann, Jorn Piel, Christophe Jacob, Bertrand Aigle, Kira J. Weissman
NATURE COMMUNICATIONS
(2022)
Article
Microbiology
Thierry Dulermo, Clara Lejeune, Ece Aybeke, Sonia Abreu, Jean Bleton, Michelle David, Ariane Deniset-Besseau, Pierre Chaminade, Annabelle Thibessard, Pierre Leblond, Marie-Joelle Virolle
Summary: Streptomyces coelicolor M145 is a model strain used to study antibiotic biosynthesis regulation in Streptomyces species. A variant of this strain was unexpectedly obtained during the deletion process of the gene encoding isocitrate lyase (sco0982). This variant produces less antibiotic and has higher lipid content compared to the original strain.
Article
Chemistry, Multidisciplinary
Abigail L. Whittock, Xuefei Ding, Xavier E. Ramirez E. Barker, Nazia Auckloo, Rebecca A. Sellers, Jack M. Woolley, Krishnan Tamareselvy, Marine Vincendet, Christophe Corre, Emma Pickwell-MacPherson, Vasilios G. Stavros
Summary: Biomimicry is playing a crucial role in researching new materials for various applications. This study focuses on a crude extract from red algae, Palmaria palmata, which contains mycosporine-like amino acids - a type of photoprotective molecules. The researchers have applied this extract onto a surface and confirmed that it retains its photoprotection and light-to-heat conversion properties. They also demonstrated how to monitor stability in real-world applications using terahertz spectroscopy.
