Article
Chemistry, Multidisciplinary
Jia-Liang Chen, Yin Yang, Tiesheng Shi, Xun-Cheng Su
Summary: We evaluated the delivery of lanthanide ions (Ln(3+)) into live cells using paramagnetic NMR spectroscopy. Free Ln(3+) ions are toxic to live cells, leading to the leakage of target proteins into the extracellular media. The citrate-Ln(3+) complex is a more efficient and gentle reagent for live cell delivery compared to free Ln(3+) form.
CHEMICAL COMMUNICATIONS
(2023)
Article
Agricultural Engineering
Lu Lin, Mengyue Gong, Yanfeng Liu, Jianghua Li, Xueqin Lv, Guocheng Du, Long Liu
Summary: In this study, the production of 2'-FL was increased through metabolic engineering. By increasing the gene copies of FutC and deleting multiple genes in competitive pathways, the titer of 2'-FL was raised to 3.3 g/L and reached 10.3 g/L in a large-scale fermentation.
BIORESOURCE TECHNOLOGY
(2022)
Article
Biotechnology & Applied Microbiology
Jianli Yu, Yang Guo, Yi Gu, Xiying Fan, Fei Li, Haipeng Song, Rui Nian, Wenshuai Liu
Summary: This study developed a novel fusion tag based on repetitive amino acid sequence motifs found in silk fibroin protein, which significantly enhanced the expression of nanobodies in Escherichia coli without affecting their affinity and thermal stability. This finding has the potential to expand the applications of nanobodies in biomedical and biotechnology fields.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2022)
Article
Biotechnology & Applied Microbiology
Bum Seok Park, Yun Hee Choi, Min Woo Kim, Beom Gi Park, Eun-Jung Kim, Jin Young Kim, Jung Hwa Kim, Byung-Gee Kim
Summary: In this study, the production of 2'-FL was significantly increased by relieving catabolite repression (CR) and optimizing the catalytic activity and solubility of the fucosyltransferase (FT) enzyme. The combination of genetic engineering and protein engineering strategies resulted in a 2.4-fold increase in 2'-FL titer. The highest titer achieved was 40 g l(-1) with a productivity of 0.55 g l(-1) h(-1) and a specific yield of 1.0 g g(-1) dry cell weight.
BIOTECHNOLOGY AND BIOENGINEERING
(2022)
Article
Food Science & Technology
Lingyan Du, Jianming Yue, Yiying Zhu, Sheng Yin
Summary: In this study, a genetic manipulation strategy was developed to construct an indigo biosynthesis cell factory in E. coli for industrial production. The over-expression of styAB gene increased the capacity of indigo biosynthesis, while the expression of tryptophanase and chaperone had different effects on indigo yield.
Article
Microbiology
Huanan Li, Lihong Huang, Yuandi Yu, Xingxing Ren, Bo Li, Jiahao Zhang, Ming Liao, Wenbao Qi
Summary: In this study, replication-competent recombinant influenza viruses were generated by inserting a Strep-Tag into PB2, and host factors interacting with viral RdRp bearing a 627 K or 627E PB2 were purified. Through mass spectrometry and network analysis, 57 host factors interacting with RdRp were identified. These proteins may act as host range and virulence determinants of influenza virus.
VETERINARY MICROBIOLOGY
(2021)
Article
Veterinary Sciences
Lei Yin, Qianwen Li, Zeping Wang, Jian Tu, Ying Shao, Xiangjun Song, Xiaocheng Pan, Kezong Qi
Summary: The E. coli type III secretion system 2 (ETT2) plays a crucial role in the virulence of avian pathogenic E. coli (APEC), with ygeG identified as a chaperone protein that affects biofilm formation, cell adherence, invasion, and bacterial virulence. Deletion of ygeG also enhances bacterial virulence in chicken infection models, regulating genes involved in biofilm formation, outer membrane proteins, fimbrial genes, and virulence effector proteins.
RESEARCH IN VETERINARY SCIENCE
(2021)
Article
Biotechnology & Applied Microbiology
Zhendong Li, Cong Gao, Chao Ye, Liang Guo, Jia Liu, Xiulai Chen, Wei Song, Jing Wu, Liming Liu
Summary: With the aim of increasing the production efficiency of microbial shikimate, researchers identified ten key target genes and used them to promote metabolic flux towards shikimate biosynthesis in the tryptophan-overproducing strain Escherichia coli TRP0. By deleting quinate dehydrogenase and introducing the shikimate dehydrogenase mutant AroET61W/L241I, the byproducts quinate and 3-dehydroshikimic acid were significantly reduced. Furthermore, the use of a high concentration shikimate responsive promoter PrpoS and dynamic regulation of the tolerance target protein ProV resulted in an increase in shikimate productivity. With these advancements, the shikimate titer reached the highest reported levels in E. coli to date.
METABOLIC ENGINEERING
(2023)
Article
Chemistry, Multidisciplinary
Charles Burridge, Christopher A. Waudby, Tomasz Wlodarski, Anais M. E. Cassaignau, Lisa D. Cabrita, John Christodoulou
Summary: This study used solution-state NMR spectroscopy to measure transverse proton relaxation rates for methyl groups in folded ribosome-nascent chain complexes, revealing interactions between the nascent chain and ribosome surface driven predominantly by electrostatics. By observing changes in these interactions as subsequent domain emerges, the impact on free energy landscapes associated with co-translational folding process can be deduced.
Article
Biochemistry & Molecular Biology
Youssef El Mouali, Milan Gerovac, Raminta Mineikaite, Joerg Vogel
Summary: FinO-domain proteins, as an emerging family of RNA-binding proteins, have diverse roles in bacterial post-transcriptional control and physiology, with the ability to act selectively or promiscuously on the same cellular RNA pool. The study extensively profiled cellular transcripts associated with the virulence plasmid-encoded FinO in Salmonella, identifying two major ligands: FinP and RepX. The research also observed cross-regulation of plasmids on the RNA level, with the FinO RBP controlling the replication of an antibiotic resistance plasmid in Salmonella.
NUCLEIC ACIDS RESEARCH
(2021)
Article
Multidisciplinary Sciences
Hisham Mazal, Marija Iljina, Inbal Riven, Gilad Haran
Summary: Recent research on AAA+ ring-shaped machines such as ClpB and Hsp104 has revealed that pore loops undergo large-amplitude fluctuations on the microsecond time scale and change their conformation in an ATP-dependent manner during substrate threading. The conformational dynamics of certain pore loops strongly correlate with disaggregation activity, suggesting they play a key role in substrate pulling. These findings support the proposal of an ultrafast Brownian-ratchet translocation mechanism that likely acts in parallel to the slower hand-over-hand process in AAA+ machines like ClpB.
Article
Chemistry, Analytical
Limin Yang, Yiping Zhang, Xingdong Dong, Xuan Wang, Mingming Li, Lei Jiang, Zhen Liu
Summary: In this study, a method for the indirect involvement of magnetic beads (MBs) in impedance aptasensing was developed. The key to this method is the construction of a Strep-Tag system integrated apta-magnetic probe, which enables the indirect attachment of the acetamiprid-binding aptamer (ABA) to Strep-Tactin-coated MBs (STMBs) for impedance measurements.
MICROCHEMICAL JOURNAL
(2023)
Article
Multidisciplinary Sciences
Lorena Ilcu, Lukas Denkhaus, Anton Brausemann, Lin Zhang, Oliver Einsle
Summary: Mono- and multiheme cytochromes c in Escherichia coli are matured through covalent attachment of heme using the complex Ccm-system. Cryo-electron microscopic structures of the heme translocation complex CcmABCD from E. coli are reported, showcasing its mechanism of shuttling heme across the membrane to the heme chaperone CcmE. ATP hydrolysis at CcmA and a conserved heme-handling motif (WxWD) at CcmC facilitate the rotation and covalent attachment of heme.
NATURE COMMUNICATIONS
(2023)
Article
Agriculture, Multidisciplinary
Daoyi Guo, Shaoting Wu, Xiao Fu, Hong Pan
Summary: This study established a biosynthetic pathway for the production of methyl cinnamate from glucose in engineered Escherichia coli, and successfully increased the titer of methyl cinnamate production.
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
(2022)
Article
Biochemistry & Molecular Biology
Subbaian Brindha, Yutaka Kuroda
Summary: The use of a solubility-enhancing peptide tag containing nine arginine residues can significantly improve the solubility of the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein expressed in Escherichia coli. This discovery has the potential to accelerate the development of anti-COVID-19 therapeutics. The folded RBD-C9R demonstrated binding activity with the host cell receptor and elicited a strong immune response in mice.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Article
Biochemistry & Molecular Biology
Patrick Pilak, Andre Schiefner, Judith Seiboth, Johannes Oehrlein, Arne Skerra
Review
Biotechnology & Applied Microbiology
Friedrich-Christian Deuschle, Elena Ilyukhina, Arne Skerra
Summary: Anticalin proteins are engineered versions of lipocalins that form a new class of clinical-stage biopharmaceuticals. Through combinatorial gene libraries and molecular selection techniques, the natural ligand specificities of lipocalins have been expanded from small molecules to peptides and proteins, leading to the development of promising Anticalin lead candidates in various disease areas. Anticalins offer an alternative to antibodies with potentially superior features as next-generation biologics, with examples of drug candidates under preclinical and clinical development.
EXPERT OPINION ON BIOLOGICAL THERAPY
(2021)
Article
Hematology
Marco Mannes, Arthur Dopler, Oliver Zolk, Sophia J. Lang, Rebecca Halbgebauer, Britta Hoechsmann, Arne Skerra, Christian K. Braun, Markus Huber-Lang, Hubert Schrezenmeier, Christoph Q. Schmidt
Summary: Use of C5 inhibitor eculizumab to block the terminal complement pathway has revolutionized clinical management of complement-mediated diseases. New findings show that C3/C5 convertases function differently than previously thought, with C3 inhibition not necessarily leading to C5 inhibition. Surface-deposited C4b alone can recruit and activate C5, and only C5 bound on C3b/C4b-decorated surfaces requires surface fixation for activation.
Article
Biochemistry & Molecular Biology
Uli Binder, Arne Skerra
Summary: This study successfully developed a long-acting version of Tα1, which effectively improves its plasma half-life in vivo, laying the foundation for the therapeutic development of the next generation of thymosin alpha 1. By combining PASylation technology with RimJ enzyme-mediated N-acetylation, the problems of stability, protein degradation, and low bioactivity of synthetic peptide drugs were addressed.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2021)
Article
Biochemistry & Molecular Biology
Andreas Eichinger, Irmgard Neumaier, Arne Skerra
Summary: Bovine butyrophilin (BTN1A1) is a glycoprotein found on milk fat globules that acts as a hydrophilic coat protein. Its structure differs from human immune receptors but shows similarity to human myelin oligodendrocyte glycoprotein. This discovery may provide insights into the correlation between neurological autoimmune diseases and cow milk consumption.
BIOLOGICAL CHEMISTRY
(2021)
Article
Oncology
Caroline Werner, Stefan Stangl, Lukas Salvermoser, Melissa Schwab, Maxim Shevtsov, Alexia Xanthopoulos, Fei Wang, Ali Bashiri Dezfouli, Dennis Tholke, Christian Ostheimer, Daniel Medenwald, Martin Windberg, Matthias Bache, Martin Schlapschy, Arne Skerra, Gabriele Multhoff
Summary: Tumor-specific biomarkers, such as Hsp70 in liquid biopsies, are useful for tumor detection, monitoring of responses, and outcome prediction. The compHsp70 ELISA, using two monoclonal antibodies, is a sensitive and reliable tool for measuring free and vesicular Hsp70 levels in tumor patients' circulation, serving as a potential biomarker for risk assessment and therapeutic monitoring.
Article
Chemistry, Multidisciplinary
Jonas Schilz, Charlotte Clement, Franziska Greiner, Arne Skerra
Summary: A novel strategy for the efficient and gentle affinity purification of PASylated proteins is reported, which allows direct one-step purification from cell extracts or culture supernatants while avoiding harsh elution conditions. This method can improve the efficiency of biological drug development.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2022)
Article
Biochemistry & Molecular Biology
Andreas Eichinger, Sabine Rauth, Dominik Hinz, Anna Feuerbach, Arne Skerra
Summary: In this study, the structural analysis of two Anticalin(R) proteins that tightly bind A beta(40) peptide, a key player in Alzheimer's disease, were described. These proteins, engineered based on human lipocalin 2, show compact single-domain binding properties and offer a potential alternative to antibodies. Both crystal structures of the engineered anticalins revealed their ability to recognize the same epitope in the middle of the beta-amyloid peptide. This finding suggests that anticalins may play a significant role in the pathophysiology of Alzheimer's disease.
BIOLOGICAL CHEMISTRY
(2022)
Article
Gastroenterology & Hepatology
Anita Annahazi, Thomas Erwin Berger, Ihsan Ekin Demir, Florian Zeller, Michael Muller, Markus Anneser, Arne Skerra, Klaus Michel, Michael Schemann
Summary: This study demonstrated for the first time the presence of functional 5-HT1P receptors on human submucous neurons and found no evidence for a role of 5-HT4 or 5-HT7 receptors in the postsynaptic activation of human submucous neurons by 5-HT.
NEUROGASTROENTEROLOGY AND MOTILITY
(2022)
Article
Biochemistry & Molecular Biology
Laura Job, Anja Kohler, Andreas Eichinger, Mauricio Testanera, Benjamin Escher, Franz Worek, Arne Skerra
Summary: Neurotoxic organophosphorus compounds (OPs) can be extremely dangerous when misused. A potential treatment involves using an enzyme to break down the nerve agent into harmless substances. The bacterium Brevundimonas diminuta (BdPTE) has been studied for its potential in this area. Engineering studies have led to improved versions of BdPTE with high catalytic efficiency. Unexpectedly, the crystal structure of a highly active variant revealed a new conformation in the active site region, leading to further amino acid exchanges and optimization of the enzyme for OP nerve agent detoxification.
Article
Biochemistry & Molecular Biology
Lea Naestle, Friedrich-Christian Deuschle, Volker Morath, Arne Skerra
Summary: The transferrin receptor (TfR) plays a role in transporting substances across the blood-brain barrier, offering a non-invasive way to deliver therapeutics to the brain. Researchers have identified a specific lipocalin, called FerryCalin, which binds to murine TfR and shows potential as a vehicle for delivering biopharmaceuticals to the brain. Through affinity maturation, lipocalin variants were engineered to bind to TfR in a non-competitive manner, with FerryCalin exhibiting a dissociation constant of 3.8 nM. This research provides insight into the potential use of FerryCalin or its mutants for brain drug delivery.
Article
Business
Stefanie Weniger, Svenja Jarchow, Oleg Nenadic
Summary: This study investigates the factors influencing entrepreneurs' preferences for corporate venture capital (CVC) investors compared to independent venture capital (IVC) investors. The results highlight the importance of resource needs, such as marketing resources and access to the corporate network, in the decision-making process. The study also challenges the assumption that CVC investments are seen as a pathway to acquisitions, and finds that entrepreneurs pursuing an IPO are less likely to prefer CVC.
JOURNAL OF SMALL BUSINESS AND ENTERPRISE DEVELOPMENT
(2023)
Article
Business
Svenja Jarchow, Christoph Kaserer, Henry Keppler
Summary: The study finds that family control positively impacts firm performance and valuation, particularly during times of crisis. Family firms in Germany exhibited significant outperformance in terms of ROA and Tobin's Q during the crisis years 2008-2010, especially when family influence in terms of equity ownership was stronger. These family firms also showed resilience by reducing leverage without sacrificing future cash flows and increasing capital expenditures and employment.
EURASIAN BUSINESS REVIEW
(2023)
Article
Biochemical Research Methods
Elena Jerschke, Andreas Eichinger, Arne Skerra
Summary: Using Anticalin technology, a lipocalin protein named Colchicalin has been engineered with high affinity for the toxic plant alkaloid colchicine, offering potential as an antidote and for sensitive detection in biological samples. The crystal structure of unliganded Colchicalin and its complex with colchicine were compared, revealing significant rearrangements in four structurally variable loops. Upon binding to colchicine, a dramatic shift and side-chain flip occur in loop #3 and Phe71, respectively, creating space in the ligand pocket. These findings demonstrate a conformational adaptation mechanism similar to that observed in antibodies, highlighting the general similarity between antigen/ligand binding by immunoglobulins and lipocalins.
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS
(2023)
Article
Biochemistry & Molecular Biology
Laura Job, Anja Koehler, Mauricio Testanera, Benjamin Escher, Franz Worek, Arne Skerra
Summary: In this study, the phosphotriesterase of Brevundimonas diminuta (BdPTE) was rationally engineered to improve its hydrolysis efficiency of malaoxon, a common pesticide metabolite. The mutant BdPTE(VRNVVLARY) showed significantly increased hydrolysis rate and improved properties including expression yield, thermal stability, and oxidation resistance. This mutant BdPTE could be a promising candidate for developing a biocatalytic antidote for the detoxification of common pesticides and related organophosphorus compounds.
PROTEIN ENGINEERING DESIGN & SELECTION
(2023)