Article
Microbiology
Emmanuele Severi, Mariana Bunoro Batista, Adelie Lannoy, Phillip J. Stansfeld, Tracy Palmer
Summary: The twin arginine transport (Tat) pathway is a mechanism in prokaryotes and chloroplasts that exports folded proteins across membranes. Through mutagenesis experiments, it has been found that a binding site located in the sixth transmembrane helix is crucial for the binding of TatA.
Article
Multidisciplinary Sciences
May N. Taw, Jason T. Boock, Belen Sotomayor, Daniel Kim, Mark A. Rocco, Dujduan Waraho-Zhmayev, Matthew P. DeLisa
Summary: The study suggests that the Tat translocase may use molecular chaperones to monitor the conformational status of its substrates, with the TatB component playing a key role in sensing the folding state of Tat substrates.
SCIENTIFIC REPORTS
(2022)
Article
Immunology
Elia Riquelme, Sabina Omarova, Berengere Ize, David O'Callaghan
Summary: Bacteria use the Tat system to export folded proteins. The Tat system is essential for the pathogen Brucella spp. By bioinformatic screening, 28 B. suis proteins with putative Tat signal sequences were identified.
INFECTION AND IMMUNITY
(2023)
Article
Biochemistry & Molecular Biology
Binhan Hao, Wenjie Zhou, Steven M. Theg
Summary: The TMH lengths of TatA and TatB are evolutionarily tuned to 15 amino acids, with activity dropping off following any modification of this length. Even with as few as 11 residues, TatA and TatB can still insert into the membrane bilayer, albeit with a decline in membrane integrity. These findings support a model of Tat transport utilizing toroidal pores formed when the membrane bilayer reaches a critical threshold.
JOURNAL OF BIOLOGICAL CHEMISTRY
(2022)
Article
Biochemistry & Molecular Biology
Wenjie Zhou, Binhan Hao, Terry M. Bricker, Steven M. Theg
Summary: The twin arginine translocation (Tat) pathway plays an important role in protein transport in bacteria, archaea, and plants. By using a real-time luminescence assay, this study reveals that both Delta psi and Delta pH contribute energetically to Tat transport in Escherichia coli spheroplasts.
JOURNAL OF BIOLOGICAL CHEMISTRY
(2023)
Article
Biotechnology & Applied Microbiology
Cuiping Pang, Song Liu, Guoqiang Zhang, Jingwen Zhou, Guocheng Du, Jianghua Li
Summary: Several strategies were adopted to improve the extracellular secretion of LOX from Escherichia coli, including signal peptides and cell wall permeability. Despite marginal improvements in LOX secretion capacity with signal peptides, the extracellular LOX yield was not sufficient for industrial production. An autolysis system based on the bacteriophage lysis gene phi X174-E was constructed to enhance extracellular protein production, leading to an optimized extracellular LOX yield of 368 +/- 1.4 U mL(-1) in a 5-L bioreactor.
MICROBIAL CELL FACTORIES
(2022)
Review
Physiology
Sharbani Kaushik, Haoze He, Ross E. Dalbey
Summary: This review summarizes the crucial role of signal peptides in protein export and membrane protein insertion, as well as their function as key factors for protein targeting and translocation machinery activation. The importance of signal peptide removal and degradation, as well as the emerging research on signal peptidases as novel targets for antibiotic development, are also discussed.
FRONTIERS IN PHYSIOLOGY
(2022)
Article
Biochemical Research Methods
Joshua Diaz, Miles Pietsch, Marissa Davila, Gerardo Jaimes, Alexis Hudson, Jean-Philippe Pellois
Summary: This study examines the impact of payload conjugation on the cell-penetrating activity of dfTAT. The results show that smaller payloads have little effect on dfTAT's cell delivery activity, while larger payloads significantly reduce both its endocytic uptake and endosomal escape efficiency.
BIOCONJUGATE CHEMISTRY
(2023)
Article
Microbiology
Manasa Bharathwaj, Chaille T. Webb, Grishma Vadlamani, Christopher J. Stubenrauch, Tracy Palmer, Trevor Lithgow
Summary: The cell envelope of Gram-negative bacteria consists of two membranes surrounding the periplasm and peptidoglycan layer. Beta-lactam antibiotics target the periplasmic penicillin-binding proteins that synthesize peptidoglycan, resulting in cell death. Bacteria can resist the effects of beta-lactam drugs by populating the periplasmic space with beta-lactamases, which are spread through lateral transfer of genes encoding them. Understanding the limitations and evolution of beta-lactamases like BKC-1 is important for controlling the spread of antimicrobial resistance.
Article
Biochemistry & Molecular Biology
Elif Eren, Norman R. Watts, Davide Randazzo, Ira Palmer, Dan L. Sackett, Paul T. Wingfield
Summary: HIV-1 Rev is a regulatory protein that plays a crucial role in transporting viral mRNA from the nucleus to the cytoplasm. This study reveals the structure of a Rev-tubulin complex and demonstrates its ability to destabilize microtubules, thereby modulating the cellular cytoskeleton.
Article
Biochemical Research Methods
Joseph Ryan Polli, Joseph P. Balthasar
Summary: Cell penetrating peptides conjugated to delivery vehicles can enhance the delivery of macromolecules. This study investigates the effects of conjugation to cell penetrating and endosomal escape peptides on the pharmacokinetics and distribution of an anti-carcinoembryonic antigen monoclonal antibody in a mouse model of colorectal cancer. The results show that peptide conjugation decreases exposure in tissues and tumors, increases plasma clearance, and distribution volume. The isoelectric point of the peptide conjugates affects the pharmacokinetic parameters.
BIOCONJUGATE CHEMISTRY
(2022)
Article
Microbiology
Jinfeng Shao, Gunjan Arora, Javier Manzella-Lapeira, Joseph A. Brzostowski, Sanjay A. Desai
Summary: This study developed a new method called RISE, which enables continuous nondestructive tracking of antigen exposure on infected cells. Using RISE, researchers tracked the membrane insertion of a parasite antigen called CLAG3 in infected red blood cells. They found that CLAG3 insertion occurs at specific parasite stages and is required for the formation of a nutrient uptake channel. Furthermore, they defined constraints on protein insertion at the host cell membrane by varying the size and charge of the extracellular domain.
Article
Chemistry, Analytical
Heng Tang, Junran Peng, Xin Jiang, Shuang Peng, Fang Wang, Xiaocheng Weng, Xiang Zhou
Summary: We presented an efficient RNA imaging strategy based on CRISPR-Cas and Tat peptide with TRAP-tag. By fusing modified CRISPR-Cas RNA hairpin binding proteins with a Tat peptide array that recruits modified RNA aptamers, this strategy allows highly precise and efficient visualization of endogenous RNA in cells. The modular design of CRISPR-TRAP-tag enables the substitution of sgRNAs, RNA hairpin binding proteins, and aptamers to optimize imaging quality and live cell affinity. We successfully visualized exogenous GCN4, endogenous mRNA MUC4, and lncRNA SatIII in single live cells using CRISPR-TRAP-tag.
Article
Biotechnology & Applied Microbiology
Jinyang Li, Yidi Wang, Kun Yang, Xiaolu Wang, Yuan Wang, Honglian Zhang, Huoqing Huang, Xiaoyun Su, Bin Yao, Huiying Luo, Xing Qin
Summary: In this study, an efficient protein expression system was established in Myceliophthora thermophila, enabling high-level expression and secretion of various proteins. The system was stable and could be used for the production of different enzymes, expanding the biotechnological potential of M. thermophila.
MICROBIAL CELL FACTORIES
(2023)
Article
Biochemical Research Methods
David Romero-Suarez, Tune Wulff, Yixin Rong, Tadas Jakociunas, Satoshi Yuzawa, Jay D. Keasling, Michael K. Jensen
Summary: Protein misfolding and aggregation are associated with neurodegenerative diseases and suboptimal protein expression in biotechnology. A promoter-reporter system based on yeast response to misfolded protein was developed to monitor protein aggregates, showing a 12-fold output dynamic range and the ability to distinguish different protein states. Optimization of promoters and culture conditions improved the performance of the reporter system.
ACS SYNTHETIC BIOLOGY
(2021)
Article
Biotechnology & Applied Microbiology
Jingqi Chen, Oscar P. Kuipers
Summary: This study reinforces the importance of a well-installed LanBTC complex in the cell membrane for the biosynthesis and transport of lanthipeptides. The recruitment of SpaT from the peripheral cell membrane to the cell poles by the NisBC complex in L. lactis was observed, which may explain the mechanism by which the secretion of the premature peptide is prevented.
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
(2022)
Article
Biochemistry & Molecular Biology
Ruben Cebrian, Ricardo Lucas, Maria Victoria Fernandez-Cantos, Koen Slot, Pablo Penalver, Marta Martinez-Garcia, Antonio Parraga-Leo, Maria Violante de Paz, Federico Garcia, Oscar P. P. Kuipers, Juan Carlos Morales
Summary: In this study, a family of aminoalkyl resveratrol derivatives with antibacterial activity inspired by cationic amphipathic antimicrobial peptides was discovered. These molecules showed good antimicrobial activity against anaerobic bacteria and exhibited synergistic effects with traditional antibiotics against Gram-negative pathogens. The mechanism of action was found to be at the membrane level and no cross-resistance was observed, although some cytotoxicity was detected.
JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
(2023)
Article
Biochemical Research Methods
Patricia Arias-Orozco, Yunhai Yi, Fleur Ruijne, Ruben Cebrian, Oscar P. Kuipers
Summary: This study explores the capabilities and limitations of the ProcM-like enzyme SyncM in synthesizing lanthipeptides. By modifying the substrate structure, researchers designed the first single-SyncA-based antimicrobial. The findings of this study will aid future bioengineering research and broaden the application scope of SyncM.
ACS SYNTHETIC BIOLOGY
(2022)
Article
Microbiology
Fangfang Liu, Auke J. J. van Heel, Jingqi Chen, Oscar P. P. Kuipers
Summary: Circular bacteriocins, a subgroup of ribosomally synthesized and post-translationally modified peptides, exhibit high stability and resistance to degradation, making them potential candidates for various applications. However, little is known about their biosynthesis and bioactivity. In this study, an efficient cloning and expression system for circularin A was developed, allowing for successful production and characterization of this circular peptide. Mutagenesis studies revealed the effects of specific residue mutations on bioactivity. The production conditions for bacteriocin were also optimized. Overall, this research provides valuable insights into circularin A biosynthesis and paves the way for further study.
FRONTIERS IN MICROBIOLOGY
(2022)
Article
Biotechnology & Applied Microbiology
Luiza P. Morawska, Oscar P. Kuipers
Summary: Bacillus subtilis, a soil-dwelling bacterium, can interact with various other microorganisms. A study found that it can transfer plasmids to lactic acid bacteria, without involving conjugation or activating the competent state. The study also discovered non-conjugative cell-to-cell plasmid transfer in non-competent strains of Lactococcus lactis sp. cremoris. This suggests that cell-to-cell transformation is a common form of horizontal gene transfer and could be used as an alternative tool for natural strain improvement.
MICROBIAL BIOTECHNOLOGY
(2023)
Review
Biochemistry & Molecular Biology
Silja Mordhorst, Fleur Ruijne, Anna L. Vagstad, Oscar P. Kuipers, Joern Piel
Summary: Peptide natural products are valuable structures for drug design and engineering. This review focuses on comparing structural features between nonribosomal and ribosomally synthesised and posttranslationally modified peptides, and highlights synthetic biology strategies for emulating nonribosomal peptides using ribosomal pathways.
RSC CHEMICAL BIOLOGY
(2023)
Article
Microbiology
Micaela Mossop, Luca Robinson, Jhih-Hang Jiang, Anton Y. Peleg, Luke Blakeway, Nenad Macesic, Audrey Perry, Stephen Bourke, Fatima R. Ulhuq, Tracy Palmer
Summary: One third of CF patients in the UK are co-infected with both Staphylococcus aureus and Pseudomonas aeruginosa, which contribute to lung tissue destruction and respiratory failure. This study aims to characterize twenty-five clinical S. aureus isolates from CF patients, including those with mono- and coinfection with P. aeruginosa, using molecular and phenotypic tools.
JOURNAL OF MEDICAL MICROBIOLOGY
(2023)
Article
Microbiology
Emmanuele Severi, Mariana Bunoro Batista, Adelie Lannoy, Phillip J. Stansfeld, Tracy Palmer
Summary: The twin arginine transport (Tat) pathway is a mechanism in prokaryotes and chloroplasts that exports folded proteins across membranes. Through mutagenesis experiments, it has been found that a binding site located in the sixth transmembrane helix is crucial for the binding of TatA.
Article
Biochemical Research Methods
Fangfang Liu, Auke J. van Heel, Oscar P. Kuipers
Summary: In this study, the leader and terminal residue requirements for the biosynthesis of circularin A were investigated through systematic mutational analyses. The results showed that a leader with only one Met residue is sufficient for producing mature circularin A. Short-sidechain hydrophobic residues at positions Val1 and Ala2 of the N-terminus and an aromatic residue at the C-terminal Tyr69 position are required for the formation of active peptide derivatives. The requirements for the residue at position Ala68 are more relaxed. Overall, this study provides new insights into the biosynthesis of circular bacteriocins and their potential applications in modified peptide engineering.
ACS SYNTHETIC BIOLOGY
(2023)
Article
Genetics & Heredity
Kieran Bowran, Stephen R. Garrett, Arnoud H. M. van Vliet, Tracy Palmer
Summary: The type VIIb protein secretion system (T7SSb) is a competitive mechanism found in Bacillota bacteria. EssC, a crucial component of T7SSb, plays a significant role in substrate recognition. Analysis of Listeria monocytogenes genomes reveals that EssC has seven sequence variants, each associated with specific candidate substrate proteins.
MICROBIAL GENOMICS
(2023)
Article
Chemistry, Multidisciplinary
Roderick Corstiaan Abraham Versloot, Patricia Arias-Orozco, Matthijs Jonathan Tadema, Florian Leonardus Rudolfus Lucas, Xinghong Zhao, Siewert J. J. Marrink, Oscar Paul Kuipers, Giovanni Maglia
Summary: Mass spectrometry cannot distinguish between molecules with the same mass-to-charge ratio, but nanopore technology can differentiate diastereomeric and enantiomeric differences in peptides that cannot be observed by conventional MS analysis. The resolution in nanopore analysis is likely due to multiple chiral interactions during peptide transport across the nanopore, similar to chiral chromatography. Nanopore technology also offers possibilities for the rapid and cost-effective analysis of peptides, including those that cannot be easily differentiated by mass spectrometry.
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
(2023)
Review
Microbiology
Yuxin Fu, Yanli Xu, Fleur Ruijne, Oscar P. Kuipers
Summary: Potent non-ribosomal antimicrobial peptides can be mimicked by combining posttranslational modifications of gene-encoded lanthipeptides, allowing screening of mutant libraries. Synthetic biology provides new avenues in peptide engineering to produce new-to-nature peptides with enhanced bioactivities. The modularity of lanthipeptides enables their engineering and screening in a high-throughput manner, making them promising tools for the production and screening of novel peptides, including mimics of potent non-ribosomally produced antimicrobial peptides.
FEMS MICROBIOLOGY REVIEWS
(2023)
Article
Microbiology
Longcheng Guo, Joseph Wambui, Chenhui Wang, Francis Muchaamba, Maria Victoria Fernandez-Cantos, Jaap Broos, Taurai Tasara, Oscar P. Kuipers, Roger Stephan
Summary: Cesin, a novel variant of nisin, efficiently kills bacteria by inhibiting cell wall synthesis without pore formation. It demonstrates a broad and potent antimicrobial profile, attributed to lipid II binding and electrostatic interactions with teichoic acids. The stability tests show that cesin is highly stable under different environmental conditions.
MICROBIOLOGY SPECTRUM
(2023)
Article
Chemistry, Medicinal
Ruben Cebrian, Efres Belmonte-Reche, Valentina Pirota, Anne de Jong, Juan Carlos Morales, Mauro Freccero, Filippo Doria, Oscar P. Kuipers
Summary: Guanidine DNA quadruplex structures play a critical role in regulating important biological activities in cells and their potential as therapeutic drug targets in bacteria is still underexplored. This study tested the biological activity of a G4-DNA ligand library against different types of bacteria and found that the activity varied depending on the bacterial group, possibly due to the different prevalence and role of G4-DNA structures in each group.
JOURNAL OF MEDICINAL CHEMISTRY
(2022)
Article
Medicine, Research & Experimental
Luiza P. Morawska, Jhonatan A. Hernandez-Valdes, Oscar P. Kuipers
Summary: Microbial communities are constantly exposed to unpredictable changes in their environment, leading microorganisms to develop the ability to switch phenotypes quickly. Bet-hedging is a strategy in which populations diversify their phenotypes randomly to spread risks, allowing them to persist in dynamic habitats. However, this strategy may come with a fitness cost in specific environments.
WIRES MECHANISMS OF DISEASE
(2022)
Article
Biochemistry & Molecular Biology
Ankita Chadda, Alexander G. Kozlov, Binh Nguyen, Timothy M. Lohman, Eric A. Galburt
Summary: In this study, it was found that the DNA damage response in Mycobacterium tuberculosis differs from well-studied model bacteria. The DNA repair helicase UvrD1 in Mtb is activated through a redox-dependent process and is closely associated with the homo-dimeric Ku protein. Additionally, Ku protein is shown to stimulate the helicase activity of UvrD1.
JOURNAL OF MOLECULAR BIOLOGY
(2024)
