Article
Chemistry, Physical
Ritu Toor, Lysandre Hourdin, Sharvina Shanmugathasan, Pauline Lefrancois, Stephane Arbault, Veronique Lapeyre, Laurent Bouffier, Jean-Paul Douliez, Valerie Ravaine, Adeline Perro
Summary: The design of enzymatic droplet-sized reactors is an important challenge with potential applications in medical diagnostics, water purification, bioengineering, and the food industry. This study demonstrates the construction of enzymatic reactors using coacervates formed by ampholyte polymer chains stabilized with agar. The coacervates can sequester and preserve the catalytic activity of enzymes such as glucose oxidase and catalase. The study investigates the enzymatic cascade reaction within the coacervates and provides new insights into the impact of proximity and confinement on enzymatic activity.
JOURNAL OF COLLOID AND INTERFACE SCIENCE
(2023)
Article
Engineering, Environmental
Evamaria Hofmann, Lena Schmauser, Julia Neugebauer, Didier Touraud, Fabrice Gallou, Werner Kunz
Summary: A cascade reaction involving a Heck reaction and an enzyme-catalyzed reaction was conducted in different aqueous solutions, and the impact of solvent structuring was investigated. Surfactant-free microemulsions were found to be the most effective solvent for the reaction, allowing for a reduction in reaction temperature. The study also revealed that the presence of surfactants is not the main factor enabling organic reactions in water, but rather the solubility and stability of the reaction components.
CHEMICAL ENGINEERING JOURNAL
(2023)
Article
Biotechnology & Applied Microbiology
Patrick R. Harris, Martha A. Grover, Ronald W. Rousseau, Andreas S. Bommarius
Summary: This study characterized the kinetics of cephalexin synthesis and hydrolysis of the precursor PGME under different substrate concentrations. It was found that previous models were inaccurate in predicting reaction yields, and 7-ADCA was discovered to be a potent inhibitor of cephalexin hydrolysis. The study also compared different kinetic models for cephalexin synthesis and simulated a continuous stirred-tank reactor.
BIOTECHNOLOGY AND BIOENGINEERING
(2022)
Article
Nanoscience & Nanotechnology
Tien K. Nguyen, Satoshi Abe, Makoto Kasamatsu, Basudev Maity, Keitaro Yamashita, Kunio Hirata, Mariko Kojima, Takafumi Ueno
Summary: This study demonstrates the potential of engineering in-cell protein crystals to immobilize two enzymes that promote a cascade reaction, significantly enhancing the reactivity of the reaction. This higher reactivity is attributed to the efficient diffusion of substrate and intermediate through extended channels designed within the nanoporous crystal.
ACS APPLIED NANO MATERIALS
(2021)
Article
Engineering, Chemical
Xiaoyan Chen, Fan Wang, Qiang Yu, Shijie Liu, Wen Wang, Yu Zhang, Zhongming Wang, Zhenhong Yuan
Summary: The study applied a one pot cascade enzymatic catalysis for u-allulose synthesis using GI and DAE enzymes, finding that increasing enzyme loading and the molar ratios of GI vs DAE could enhance the u-allulose yield. Optimal enzyme loadings varied with different substrate concentrations.
CHEMICAL ENGINEERING SCIENCE
(2022)
Article
Chemistry, Physical
Dylan K. Courtney, Yun Su, Tyler B. Jacobson, Daven B. Khana, Aditya Ailiani, Daniel Amador-Noguez, Brian F. Pfleger
Summary: In this study, researchers have reconstituted the reversed s-oxidation (R-sox) pathway in vitro and demonstrated that by controlling the ratio of three enzymes and the ratio of NADH and NADPH, the average chain length of the alcohol product profile can be influenced.
Article
Biotechnology & Applied Microbiology
Lisa Bretschneider, Ingeborg Heuschkel, Katja Buehler, Rohan Karande, Bruno Buehler
Summary: This study successfully constructed a strain of Pseudomonas taiwanensis that efficiently converts cyclohexane to adipic acid, laying the foundation for environmentally friendly production of adipic acid and related polymers.
METABOLIC ENGINEERING
(2022)
Article
Agricultural Engineering
Guoyan Zhang, Yingfeng An, Hossain M. Zabed, Junhua Yun, Amreesh Parvez, Mei Zhao, Cunsheng Zhang, Yuvaraj Ravikumar, Jia Li, Xianghui Qi
Summary: A novel galactitol dehydrogenase was discovered and successfully used to establish an oxidoreductive pathway for D-tagatose synthesis in Bacillus subtilis. Pathway fine-tuning and optimization of fermentation conditions resulted in higher D-tagatose production.
BIORESOURCE TECHNOLOGY
(2023)
Review
Food Science & Technology
Anshula Sharma, Gaganjot Gupta, Tawseef Ahmad, Sheikh Mansoor, Baljinder Kaur
Summary: Enzyme engineering techniques, such as directed evolution and site-directed mutagenesis, play a crucial role in enhancing catalytic properties, improving industrial efficiency, and producing novel metabolites. These strategies offer excellent opportunities to tailor existing enzymes and construct highly efficient industrial enzymes for cost-effective production of value-added products.
FOOD REVIEWS INTERNATIONAL
(2021)
Article
Agricultural Engineering
Misa Mojca Cajnko, Jost Oblak, Miha Grilc, Blaz Likozar
Summary: Enzymes play a crucial role in lignin valorization, with different enzymes involved in various elementary reaction mechanisms. Degradation of lignin depends on enzyme activity and the interaction with proteins in ways that target specific covalent bonds in lignin. Various factors influence enzymatic activity, which must be taken into consideration for optimal functionality and synthesis yields.
BIORESOURCE TECHNOLOGY
(2021)
Article
Engineering, Environmental
Hao Yuan, Shuting Yi, Lufan Jia, Sicheng Chen, Ting Guo, Tao Meng
Summary: In this study, a living cell-inspired nanoscale multienzyme complex was designed to enable efficient cascade reactions and enhance the detection sensitivity of biosensors. The systematic study of this nanocomplex can inspire further investigation into the rules of cellular catalytic behaviors.
CHEMICAL ENGINEERING JOURNAL
(2023)
Article
Environmental Sciences
Ze-Gang Dong, Fang Xu, Ellen Mitchell, Bo Long
Summary: High-level ab initio calculations and variational transition state theory were used to study the aminolysis of trifluoroacetaldehyde catalyzed by a single water molecule, which significantly reduces the reaction energy barrier. Kinetic calculations showed a wide range of rate coefficients for the reaction. The study also identified a competitive elimination pathway for trifluoroacetaldehyde under certain conditions, which may lead to the growth of secondary organic aerosols.
ATMOSPHERIC ENVIRONMENT
(2021)
Article
Nanoscience & Nanotechnology
Hyeryeong Lee, Yuna Bang, In Seop Chang
Summary: This study constructed an enzyme cascade-induced bioelectrocatalytic system by using solid binding peptide (SBP) as a molecular binder to coimmobilize invertase (INV) and flavin adenine dinucleotide (FAD)-dependent glucose dehydrogenase gamma-alpha complex (GDH γ α) on a single electrode surface. It emphasizes the importance of interenzyme orientation in electrocatalytic systems and demonstrates the efficacy of SBP fusion technology as a generic tool for developing cascade-induced direct bioelectrocatalytic systems.
ACS APPLIED MATERIALS & INTERFACES
(2023)
Article
Chemistry, Multidisciplinary
Yan Xiong, Stanislav Tsitkov, Henry Hess, Oleg Gang, Yifei Zhang
Summary: This study establishes a cascade system using reversible DNA-directed colocalization of enzymes on microspheres to directly evaluate the colocalization effect. In a regime of highly dilute microspheres of particular sizes, the colocalized cascade shows enhanced activity compared to the freely diffusing cascade. The findings provide theoretical support for the rational design of highly efficient multienzyme catalysts.
Review
Agriculture, Multidisciplinary
Jiawei Meng, Yingying Zhu, Hao Wang, Hongzhi Cao, Wanmeng Mu
Summary: Human milk oligosaccharides (HMOs) are considered the new gold standard for premium infant formula due to their unique beneficial effects for infants. HMOs are a collection of unconjugated glycans, with over 200 distinct structures identified. The production of HMOs involves glycosylation using specific glycosyltransferases and nucleotide sugar donors.
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
(2023)
Article
Multidisciplinary Sciences
Emily C. Sheppard, Sally Rogers, Nicholas J. Harmer, Richard Chahwan
SCIENTIFIC REPORTS
(2019)
Review
Biochemistry & Molecular Biology
Rhys Cutlan, Simone De Rose, Michail N. Isupov, Jennifer A. Littlechild, Nicholas J. Harmer
BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS
(2020)
Article
Multidisciplinary Sciences
Sam Barker, Sarah V. Harding, David Gray, Mark I. Richards, Helen S. Atkins, Nicholas J. Harmer
Summary: Researchers developed a high-throughput assay to screen for potential co-therapy compounds to enhance the effectiveness of ceftazidime. Through screening and investigation, they identified chloroxine as a potentially effective co-therapy, reducing the number of drug-resistant bacteria.
Article
Chemistry, Multidisciplinary
Nicholas J. Harmer, Alison M. Hill
Summary: Due to the COVID-19 pandemic, the shift to online teaching and assessment posed challenges in teaching laboratory skills and ensuring fair assessment. A solution was found in using bespoke laboratory videos and unique data sets for students, as well as utilizing R to produce student data packs to prevent collusion. Results showed no difference in student performance, with mean and median marks remaining consistent with previous years.
JOURNAL OF CHEMICAL EDUCATION
(2021)
Article
Chemistry, Multidisciplinary
Melissande Louis, Thomas Clamens, Ali Tahrioui, Florie Desriac, Sophie Rodrigues, Thibaut Rosay, Nicholas Harmer, Suraya Diaz, Magalie Barreau, Pierre-Jean Racine, Eric Kipnis, Teddy Grandjean, Julien Vieillard, Emeline Bouffartigues, Pierre Cornelis, Sylvie Chevalier, Marc G. J. Feuilloley, Olivier Lesouhaitier
Summary: Pseudomonas aeruginosa biofilms can be prevented and dispersed by the human hormone Atrial Natriuretic Peptide (hANP). hANP has a dose-dependent effect and can enhance the antibiofilm action of diverse antibiotics, leading to almost complete eradication of biofilms. It lacks toxicity and acts synergistically with antibiotics.
Article
Infectious Diseases
Jua Iwasaki, Donald D. Lorimer, Mirella Vivoli-Vega, Emily A. Kibble, Christopher S. Peacock, Jan Abendroth, Stephen J. Mayclin, David M. Dranow, Phillip G. Pierce, David Fox, Maria Lewis, Nicole M. Bzdyl, Sofie S. Kristensen, Timothy J. J. Inglis, Charlene M. Kahler, Charles S. Bond, Anja Hasenkopf, Florian Seufert, Jens Schmitz, Laura E. Marshall, Andrew E. Scott, Isobel H. Norville, Peter J. Myler, Ulrike Holzgrabe, Nicholas J. Harmer, Mitali Sarkar-Tyson
Summary: This study investigated the efficacy of small-molecule compounds targeting Burkholderia pseudomallei Mip protein (BpMip) for broad-spectrum antimicrobial activity. The results showed that two of the compounds inhibited the PPIase activity of Mip proteins from other pathogens and reduced the intracellular burden of these pathogens in vitro. These findings suggest that Mip is a potential antivirulence target and the compounds may serve as promising candidates for broad-spectrum drugs.
JOURNAL OF ANTIMICROBIAL CHEMOTHERAPY
(2022)
Article
Biochemistry & Molecular Biology
Alice R. Cross, Sumita Roy, Mirella Vivoli Vega, Martin Rejzek, Sergey A. Nepogodiev, Matthew Cliff, Debbie Salmon, Michail N. Isupov, Robert A. Field, Joann L. Prior, Nicholas J. Harmer
Summary: This study found that the orthologs StrM and CBU1838 in bacteria Streptomyces griseus and Coxiella burnetii, respectively, have similar functions to the TDP enzyme and can support the biosynthesis of TDP-rhamnose. The research also showed that there are differences in proton exchange rates at the 3 and 5 positions, and the enzymes form an active site through binding with TDP. This is important for understanding the complete biosynthetic pathways of these unique sugars.
JOURNAL OF BIOLOGICAL CHEMISTRY
(2022)
Article
Biochemistry & Molecular Biology
Sumita Roy, Mirella Vivoli Vega, Jessica R. Ames, Nicole Britten, Amy Kent, Kim Evans, Michail N. Isupov, Nicholas J. Harmer
Summary: N-acetyl-D-glucosamine (GlcNAc) is an important component in bacterial cell walls and its metabolism starts with phosphorylation to GlcNAc-6-phosphate. The ROK kinase NagK is responsible for this step in bacteria. In this study, the structure of NagK in complex with GlcNAc and ATP analog AMP-PNP was solved, revealing conformational changes and a sequential random enzyme mechanism. Molecular dynamics modeling and site-directed mutagenesis further confirmed the essential catalytic metal and base residues. Overall, this study provides comprehensive insights into the activity of ROK kinases.
JOURNAL OF BIOLOGICAL CHEMISTRY
(2023)
Article
Biochemical Research Methods
Nicolas Julian Scheuplein, Theresa Lohr, Mirella Vivoli Vega, Dyan Ankrett, Florian Seufert, Lukas Kirchner, Nicholas J. Harmer, Ulrike Holzgrabe
Summary: A fluorescence polarization assay was established to screen and develop inhibitors of BpMip. The assay showed high efficiency in screening large compound libraries and identifying potent new inhibitors.
Article
Chemistry, Multidisciplinary
Alison M. Hill, Nicholas J. Harmer
Summary: Gamification has a proven track record of improving student engagement and learning in the chemical sciences. Providing a game fiction, encouraging teamwork, and breaking games into smaller quests have been found to be particularly effective approaches. In this study, a game was created to enhance students' engagement and learning of analytical biochemistry techniques through solving a murder mystery. The game was well-received by students, who reported enjoying the session and gaining confidence in the methods involved. A revised session was developed to address feedback and increase the level of challenge.
JOURNAL OF CHEMICAL EDUCATION
(2023)
Article
Biochemistry & Molecular Biology
Ida Freda, Cecile Exertier, Anna Barile, Antonio Chaves-Sanjuan, Mirella Vivoli Vega, Michail N. Isupov, Nicholas J. Harmer, Elena Gugole, Paolo Swuec, Martino Bolognesi, Anita Scipioni, Carmelinda Savino, Martino Luigi Di Salvo, Roberto Contestabile, Beatrice Vallone, Angela Tramonti, Linda Celeste Montemiglio
Summary: Specificity in protein-DNA recognition is determined by the structural and chemical features of the targeted DNA molecule. Using cryo-EM and crystallography, we elucidated the interactions driving the DNA recognition and binding by PdxR, a bacterial transcription factor. Our findings revealed the importance of electrostatic interactions and DNA bending in the allosteric regulation of DNA binding by PdxR. This study provides detailed insights into the structure and dynamics of the PdxR-DNA complex and the regulatory mechanisms of the MocR family of transcription factors.
NUCLEIC ACIDS RESEARCH
(2023)
Article
Microbiology
Melissande Louis, Ali Tahrioui, Courtney J. Tremlett, Thomas Clamens, Jerome Leprince, Benjamin Lefranc, Eric Kipnis, Teddy Grandjean, Emeline Bouffartigues, Magalie Barreau, Florian Defontaine, Pierre Cornelis, Marc G. J. Feuilloley, Nicholas J. Harmer, Sylvie Chevalier, Olivier Lesouhaitier
Summary: Biofilms are resistant to antimicrobial drugs and host immune defenses, allowing pathogens to survive in harsh environments. Atrial Natriuretic Peptide (hANP) has been shown to have strong anti-biofilm activity against Pseudomonas aeruginosa through its binding to the AmiC protein. In this study, we evaluated the potential anti-biofilm activity of osteocrin (OSTN), an h-NPRC agonist that binds to the AmiC sensor.
Article
Biology
Adam Thomas, Rhys Cutlan, William Finnigan, Mark van der Giezen, Nicholas Harmer
COMMUNICATIONS BIOLOGY
(2019)
