Article
Engineering, Manufacturing
Jonathan W. Pegues, Michael A. Melia, Raymond Puckett, Shaun R. Whetten, Nicolas Argibay, Andrew B. Kustas
Summary: The study presents an efficient method for rapidly evaluating the properties of complex concentrated alloys using powder-based directed energy deposition additive manufacturing. Through in situ alloying, a broad range of compositions in a single metallurgical sample was synthesized, allowing for a quick assessment of the alloy's performance.
ADDITIVE MANUFACTURING
(2021)
Article
Chemistry, Multidisciplinary
Christoph Kung, Rosario Vanella, Michael A. Nash
Summary: Engineering catalytic and biophysical properties of enzymes is essential for their applications in biomedical and industrial fields. In this study, a high-throughput screening and directed evolution strategy was developed to enhance the performance of an enzyme called d-Amino acid oxidase, which is a candidate for cancer therapy. By using single-cell hydrogel encapsulation and fluorescence activated cell sorting (FACS), variants with improved substrate binding affinity were isolated and characterized. This study highlights the difficulty in predicting allosteric sites and the advantages of scalable library screening technologies for evolving catalytic enzymes.
REACTION CHEMISTRY & ENGINEERING
(2023)
Article
Biochemical Research Methods
Matthew Rienzo, Ke-Chih Lin, Kellen C. Mobilia, Eric K. Sackmann, Volker Kurz, Adam H. Navidi, Jarett King, Robert M. Onorato, Lawrence K. Chao, Tony Wu, Hanxiao Jiang, Justin K. Valley, Troy A. Lionberger, Michael D. Leavell
Summary: The high-throughput perfusion-based screening system allows real-time monitoring of microbial strain productivity to improve small molecule secretion efficiency. By applying whole-genome random mutagenesis, peak specific productivities of high-productivity strains can be substantially increased, leading to time savings in screening processes.
Review
Chemistry, Multidisciplinary
Romany J. McLure, Sheena E. Radford, David J. Brockwell
Summary: Directed evolution is a powerful tool for engineering new functions in biomolecules and gaining insights into protein behavior. It accelerates the process of natural evolution through multiple rounds of gene diversification and selection.
TRENDS IN CHEMISTRY
(2022)
Article
Biochemistry & Molecular Biology
Gheorghita Menghiu, Vasile Ostafe, Radivoje Prodanovic, Rainer Fischer, Raluca Ostafe
Summary: Chitinases catalyze the degradation of chitin, a polymer found in various organisms. Developing improved chitinases is important for addressing environmental concerns and potential applications of chitosan products. This study focused on creating a high-throughput screening system and selecting enhanced chitinase variants by utilizing fluorescence-activated cell sorting and mutagenesis techniques.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2021)
Article
Food Science & Technology
Xiangdong Zhao, Roland Kerpes, Thomas Becker
Summary: This study investigates the impacts of MTP configurations on S. pastorianus beer fermentation and finds that lager strains in MTPs accelerate cell reproduction and sugar consumption. The results of microscale beer fermentation in different MTP configurations differ greatly from shaking flasks and European Brewing Convention tubes. The air-tight MTP cover not only balances flavors but also maintains the integrity of endogenous carbon dioxide pressure during fermentation. Excluding air in MTP reduces fermentative activities.
EUROPEAN FOOD RESEARCH AND TECHNOLOGY
(2022)
Article
Chemistry, Multidisciplinary
Melody A. Morris, Carolyn E. Mills, Justin M. Paloni, Eric A. Miller, Hadley D. Sikes, Bradley D. Olsen
Summary: Evolution shapes diverse properties of proteins. Incorporating proteins into materials is desired for applications, but high-throughput selection techniques are lacking. This study established a high-throughput platform to assess binding affinity for ordered sensing proteins. Fusion proteins library was constructed, and selected variants showed high binding in films. The assembly of fusion proteins and the functionality of binding protein are key factors in biosensing performance. Integration of directed evolution with assembled systems is necessary for designing better materials.
Article
Oncology
Xun Qin, Zhen Ni, Jianjun Jiang, Xiguang Liu, Xiaoying Dong, Mei Li, Kai Miao, Shuan Rao, Wenqing Zhang, Kaican Cai
Summary: This study identifies the overexpression of Piezo-Type Mechanosensitive Ion Channel Component 1 (PIEZO1) in human esophageal squamous cell carcinoma (ESCC) tumors, and demonstrates that the combination of PIEZO1 antibody and monomethyl auristatin E (MMAE) can effectively kill PIEZO1 high-expressed ESCC tumor cells. It provides a new strategy for personalized therapy of ESCC.
Article
Chemistry, Multidisciplinary
Hendrik Puetz, Christoph Janknecht, Francisca Contreras, Mariia Vorobii, Tetiana Kurkina, Ulrich Schwaneberg
Summary: A sensitive high-throughput screening system has been developed to improve the efficiency of polyamide and polyurethane recycling by tailoring polyamidases through directed depolymerase evolution. With global polymer production set to exceed 400 megatons annually, the development of recycling methods that enable a climate-neutral, circular polymer economy is crucial to address environmental pollution and climate change. This study presents the first validated high-throughput screening system for optimizing polyamidases in polyamide degradation, enabling the detection of degradation products in cell-free extract and resulting in a 1.9-fold increase in turnover frequency with just one round of random mutagenesis.
ACS SUSTAINABLE CHEMISTRY & ENGINEERING
(2023)
Review
Biotechnology & Applied Microbiology
Zi-Xu Liu, Si-Ling Huang, Jin Hou, Xue-Ping Guo, Feng-Shan Wang, Ju-Zheng Sheng
Summary: Valuable polysaccharides are typically produced through fermentation using wild-type or metabolically-engineered microbial strains, with high-throughput screening techniques being used to identify cells with high levels of polysaccharide synthesis. Strategies such as fluorescence labeling, fluorescence quenching, and biosensors are effective for cell sorting in industrial strains and pathway optimization for polysaccharide synthesis. Additionally, high-throughput screening using individual whole cells is critical in the discovery and evolution of enzymes involved in polysaccharide biosynthesis, with these enzymes and their mutants being valuable catalysts for saccharide synthesis.
MICROBIAL CELL FACTORIES
(2021)
Article
Environmental Sciences
Heidi F. Hubbard, Caroline L. Ring, Tao Hong, Cara C. Henning, Daniel A. Vallero, Peter P. Egeghy, Michael-Rock Goldsmith
Summary: Ex Priori is a flexible exposure model based on Excel, used to prioritize potential exposures for chemicals with minimal data. It quickly visualizes exposure rankings from consumer product use and considers the impact of consumer use pattern changes on exposure risk.
Article
Chemistry, Multidisciplinary
Ulrich Markel, Pia Lanvers, Daniel F. Sauer, Malte Wittwer, Gaurao Dhoke, Mehdi D. Davari, Johannes Schiffels, Ulrich Schwaneberg
Summary: Enzymatic oxidative decarboxylation is a promising reaction, but efficient screening methods for the directed evolution of decarboxylases are lacking. In this study, a simple photoclick assay was developed for detecting decarboxylation products and applied in a proof-of-principle directed evolution study on the decarboxylase OleT. The assay was compatible with different OleT operation modes and identified enzyme variants with altered substrate preference. Overall, this photoclick assay shows potential to accelerate the directed evolution of OleT and other decarboxylases.
CHEMISTRY-A EUROPEAN JOURNAL
(2021)
Review
Biochemical Research Methods
Kendreze Holland, John Blazeck
Summary: This review discusses the application of the eukaryotic yeast Saccharomyces cerevisiae as a model host in whole cell biocatalytic conversions, protein evolution, and scientific inquiries into human disease pathogenesis. The review highlights the past and present genome-scale screening platforms and recent advances in biomolecule evolution and high throughput selection and screening strategies. The potential future developments in yeast high throughput engineering are also described.
JOURNAL OF BIOLOGICAL ENGINEERING
(2022)
Article
Biochemistry & Molecular Biology
Jorge A. Lerma Romero, Christian Meyners, Andreas Christmann, Lisa M. Reinbold, Anna Charalampidou, Felix Hausch, Harald Kolmar
Summary: This article describes a protein engineering strategy that identified variants with higher affinity binding of conformation-specific FKBP51 selective inhibitors. These variants will provide a better understanding of the conformational flexibility of the FKBP51 binding pocket and may enable the isolation of new selective ligands.
FRONTIERS IN MOLECULAR BIOSCIENCES
(2022)
Review
Chemistry, Multidisciplinary
Xabier Rodriguez-Martinez, Enrique Pascual-San-Jose, Mariano Campoy-Quiles
Summary: The discovery of novel high-performing materials in organic solar cells has rapidly increased efficiency, but traditional experimentation methods are unable to evaluate the vast catalog of materials efficiently. High-throughput experimental and computational methods are being utilized to accelerate the discovery of new materials, with machine-learning algorithms playing a key role in retrieving quantitative structure-activity relationships.
ENERGY & ENVIRONMENTAL SCIENCE
(2021)
Review
Chemistry, Multidisciplinary
Jay S. Wright, Peter J. H. Scott, Patrick G. Steel
Summary: The review focuses on the borylation of heteroarenes, with an emphasis on the influence of steric and electronic effects on regiochemical outcome. It is shown that the borylation of heterocyclic substrates is more challenging, with the position and nature of the heteroatoms significantly affecting the reactivity and regioselectivity of the reaction.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2021)
Article
Chemistry, Medicinal
John G. M. Mina, Rebecca L. Charlton, Edubiel Alpizar-Sosa, Douglas O. Escrivani, Christopher Brown, Amjed Alqaisi, Maria Paula G. Borsodi, Claudia P. Figueiredo, Emanuelle de Lima, Emily A. Dickie, Wenbin Wei, Robson Coutinho-Silva, Andy Merritt, Terry K. Smith, Michael P. Barrett, Bartira Rossi-Bergmann, Paul W. Denny, Patrick G. Steel
Summary: This study successfully identified an over-the-counter antihistamine, clemastine fumarate, as a potential candidate for the treatment of leishmaniasis. Through multiple screenings and experiments, the drug showed promising activity and effectiveness, suggesting it could be repurposed for the treatment of this disease.
ACS INFECTIOUS DISEASES
(2021)
Article
Cell Biology
Hany M. Elsheikha, Alaa T. Al-Sandaqchi, Mohammad S. R. Harun, Francesca Winterton, Ali Altharawi, Nashwa A. Elsaied, Carl W. Stevenson, William MacNaughtan, John G. M. Mina, Paul W. Denny, Gianfelice Cinque, Ka Lung Andrew Chan
Summary: This study used FTIR microspectroscopy and synchrotron-based FTIR microspectroscopy to analyze the chemical changes associated with T. gondii infection of hBMECs. The results showed that both methods were able to reveal the chemical alterations in the infected cells. Furthermore, RNA sequencing analysis revealed significant changes in host cell genes and pathways in response to T. gondii infection.
Article
Plant Sciences
Nur Afiqah Sukiran, Susanna Pollastri, Patrick G. Steel, Marc R. Knight
Summary: Synthesized small molecules are useful for studying hormonal signaling in plant growth and as agrochemicals. A1 is a novel growth-promoting chemical that enhances growth in both shoot and root tissues by increasing dry and fresh weight. A1 acts via degradation of DELLA proteins and requires endogenous gibberellins for growth promotion in Arabidopsis.
Article
Pharmacology & Pharmacy
Exequiel O. J. Porta, Jaime A. Isern, Karunakaran Kalesh, Patrick G. Steel
Summary: In this study, a new therapeutic target for the treatment of leishmaniasis was discovered using an activity-based protein profiling strategy to investigate serine proteases (SPs) in Leishmania parasites. Significant differences in the expression levels of active Leishmania serine hydrolases (SHs) were observed throughout the parasite's life cycle and between different species. Two targetable SPs were identified in Leishmania mexicana using quantitative proteomic mass spectrometry. The results of this study provide templates for the discovery of new drugs for the treatment of leishmaniasis.
FRONTIERS IN PHARMACOLOGY
(2022)
Article
Chemistry, Multidisciplinary
Amy V. Hall, Isobel E. F. Gostick, Dmitry S. Yufit, Gloria Y. Marchant, Preyanthiny Kirubakaran, Shadrack J. Madu, Mingzhong Li, Patrick G. Steel, Jonathan W. Steed
Summary: The crystal structures and solvates of miltefosine were studied, revealing the existence of hydrates and methanol solvates. Water and solvent molecules play an important role in the stability of the zwitterionic structures.
CRYSTAL GROWTH & DESIGN
(2022)
Article
Infectious Diseases
Edubiel A. Alpizar-Sosa, Nur Raihana Binti Ithnin, Wenbin Wei, Andrew W. Pountain, Stefan K. Weidt, Anne M. Donachie, Ryan Ritchie, Emily A. Dickie, Richard J. S. Burchmore, Paul W. Denny, Michael P. Barrett
Summary: Amphotericin B resistance in Leishmania is associated with changes in sterol composition, which are caused by gene mutations. The sensitivity of the cells to the drug also varies depending on the growth medium. Some resistant lines show reduced virulence, but still retain their resistance phenotype.
PLOS NEGLECTED TROPICAL DISEASES
(2022)
Article
Immunology
Edubiel A. Alpizar-Sosa, Yasmine Kumordzi, Wenbin Wei, Phillip D. Whitfield, Michael P. Barrett, Paul W. Denny
Summary: With the global spread of the neglected tropical disease leishmaniasis, the validation of new drug targets in the causative insect vector is crucial. Using CRISPR Cas9 technology, a study reveals the essentiality of a gene and the possibility of simultaneous deletion of two genes, providing insights into the physiological process of the parasite.
FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY
(2022)
Article
Chemistry, Organic
Mingyan Ding, Jonathan A. Reuven, Andrew C. Hones, Mark A. Fox, Patrick G. Steel
Summary: The iridium catalysed C-H borylation of polyfluorinated arenes and heteroarenes occurs rapidly and efficiently. Steric parameters dominate the borylation reactions, but there is also an underlying electronic influence on reaction selectivity. The regioselectivity of borylation in fluorinated (hetero)arenes is mainly determined by electronic effects, except for ortho-borylation between two fluorine atoms where steric effects become apparent. Good agreement between predicted and observed selectivities is achieved by considering transition state energies in iridium geometries.
EUROPEAN JOURNAL OF ORGANIC CHEMISTRY
(2022)
Editorial Material
Immunology
Luiz R. O. Tosi, Paul William Denny, Camila I. De Oliveira, Jeziel D. Damasceno
FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY
(2023)
Article
Biochemistry & Molecular Biology
Zisis Koutsogiannis, John G. Mina, Christin A. Albus, Matthijs A. Kol, Joost C. M. Holthuis, Ehmke Pohl, Paul W. Denny
Summary: The study found that the ceramide synthases TgCerS1 and TgCerS2 in Toxoplasma gondii function differently, with TgCerS1 being catalytically active and TgCerS2 being inactive. While TgCerS1 is important but not indispensable for parasite growth, TgCerS2 plays a greater role in parasite fitness. This unique arrangement of ceramide synthase isoforms in Toxoplasma and other Apicomplexa highlights their unusual biology.
Article
Infectious Diseases
Nailma S. Aprigio-Santos, Carlos F. Estevez-Castro, Juan P. Macedo, Daniela F. Chame, Thiago Castro-Gomes, Mariana Santos-Cardoso, Gabriela A. Burle-Caldas, Courtney N. Covington, Patrick G. Steel, Terry K. Smith, Paul W. Denny, Santuza M. R. Teixeira
Summary: This study demonstrates the essential role of IPC synthase (IPCS), an enzyme involved in the synthesis of inositol-phosphorylceramide (IPC), in the growth and differentiation of Trypanosoma cruzi. The disruption of IPCS gene negatively affects the parasite's ability to differentiate into infective forms and establish an infection in vivo. These findings suggest that IPCS could be a potential target for drug development against Chagas disease.
PLOS NEGLECTED TROPICAL DISEASES
(2023)
Article
Microbiology
Exequiel O. Porta, Liqian Gao, Paul W. Denny, Patrick G. Steel, Karunakaran Kalesh
Summary: This study systematically characterizes the perturbing effect of HSP90 inhibition on the phosphoproteome of Leishmania mexicana, revealing distinct molecular effects in different life cycle stages. The findings provide insight into selective molecular targeting in the therapeutically relevant amastigote stage.
MICROBIOLOGY SPECTRUM
(2023)
Article
Chemistry, Multidisciplinary
Elizabeth V. Bedwell, Flavio da Silva Emery, Giuliano C. Clososki, Patrick G. Steel
Summary: Heterocycles are important in fragment-based drug discovery due to their prevalence in biologically active compounds. This study describes the synthesis of 5-halo-1H-pyrazolo[3,4-c]pyridine scaffolds and demonstrates how these compounds can be selectively elaborated along multiple growth-vectors. The findings highlight the potential utility of pyrazolo[3,4-c]pyridines in fragment-based drug discovery.
Article
Chemistry, Organic
Maria Schwarz, Rebecca F. M. Eno, Stefanie Freitag-Pohl, Christopher R. Coxon, Hannah E. Straker, David J. Wortley, David J. Hughes, Glynn Mitchell, Jenny Moore, Ian Cummins, Nawaporn Onkokesung, Melissa Brazier-Hicks, Robert Edwards, Ehmke Pohl, Patrick G. Steel
Summary: The study identified AmGSTF1 as a functional biomarker of MHR in black-grass and further explored its role in MHR using a combination of chemical and structural biology. It was found that the inhibitory activity of NBD-Cl is associated with the occlusion of the active site and inhibition of substrate binding, while flavonoids showed high affinity for AmGSTF1 and could reverse herbicide resistance in MHR black-grass.
ORGANIC & BIOMOLECULAR CHEMISTRY
(2021)
