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Glycosylation plays a pivotal role in tuning the folding and function of proteins. Because most human therapeutic proteins are glycosylated, understanding and controlling glycosylation is important for the design, optimization, and manufacture of biopharmaceuticals. Unfortunately, natural eukaryotic glycosylation pathways are complex and often produce heterogeneous glycan patterns, making the production of glycoproteins with chemically precise and homogeneous glycan structures difficult. To overcome these limitations, bacterial glycoengineering has emerged as a simple, cost-effective, and scalable approach to produce designer glycoprotein therapeutics and vaccines in which the glycan structures are engineered to reduce heterogeneity and improve biological and biophysical attributes of the protein. Here, we discuss recent advances in bacterial cell-based and cell-free glycoengineering that have enabled the production of biopharmaceutical glycoproteins with customized glycan structures.
Assuntos
Bactérias , Glicoproteínas , Glicosilação , Humanos , Bactérias/metabolismo , Bactérias/genética , Glicoproteínas/metabolismo , Glicoproteínas/química , Polissacarídeos/metabolismo , Polissacarídeos/química , Sistema Livre de Células , Engenharia de Proteínas/métodos , Produtos Biológicos/metabolismo , AnimaisRESUMO
The gut microbiome possesses numerous biochemical enzymes that biosynthesize metabolites that impact human health. Bile acids comprise a diverse collection of metabolites that have important roles in metabolism and immunity. The gut microbiota-associated enzyme that is responsible for the gateway reaction in bile acid metabolism is bile salt hydrolase (BSH), which controls the host's overall bile acid pool. Despite the critical role of these enzymes, the ability to profile their activities and substrate preferences remains challenging due to the complexity of the gut microbiota, whose metaproteome includes an immense diversity of protein classes. Using a systems biochemistry approach employing activity-based probes, we have identified gut microbiota-associated BSHs that exhibit distinct substrate preferences, revealing that different microbes contribute to the diversity of the host bile acid pool. We envision that this chemoproteomic approach will reveal how secondary bile acid metabolism controlled by BSHs contributes to the etiology of various inflammatory diseases.
RESUMO
The gut microbiome possesses numerous biochemical enzymes that biosynthesize metabolites that impact human health. Bile acids comprise a diverse collection of metabolites that have important roles in metabolism and immunity. The gut microbiota-associated enzyme that is responsible for the gateway reaction in bile acid metabolism is bile salt hydrolase (BSH), which controls the host's overall bile acid pool. Despite the critical role of these enzymes, the ability to profile their activities and substrate preferences remains challenging due to the complexity of the gut microbiota, whose metaproteome includes an immense diversity of protein classes. Using a systems biochemistry approach employing activity-based probes, we have identified gut microbiota-associated BSHs that exhibit distinct substrate preferences, revealing that different microbes contribute to the diversity of the host bile acid pool. We envision that this chemoproteomic approach will reveal how secondary bile acid metabolism controlled by BSHs contributes to the etiology of various inflammatory diseases.
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Main group chemistry is often considered less "dynamic" than transition metal (TM) chemistry because of predictable VSEPR-based central atom geometries, relatively slower redox switching and lack of electronic d-d transitions. However, we delineate what has been made possible with main group chemistry to give it its proper due and up-to-date treatment. The huge untapped potential regarding photophysical properties and functioning hereby spurred us to review a range of corrole reports addressing primarily photophysical trends, synthetic aspects, and important guidelines regarding substitution and inorganic principles. We also look at Ag and Au systems and also consider substitutions such as CF3, halogens, additives and also counterions. Throughout, as well as at the end of this review, we suggest various future directions; further future industrial catalytic and health science research is encouraged.
RESUMO
We describe the pertinent research steps and analysis, many of which are chemical, to achieve a novel molecular probe for glutathione (GSH) which has been published and patented based on two recent articles: "Exceptional time response, stability and selectivity in doubly-activated phenyl selenium-based glutathione-selective platform" and "Enhanced Doubly Activated Dual Emission Fluorescent Probes for Selective Imaging of Glutathione or Cysteine in Living Systems" (Kim et al., 2015; Mulay et al., 2018). The papers involve coumarin probes. Reaction/detection unfolds with aminothiol attack at an electrophilic ring carbon position. An adjacent -CHO group is heavily involved in resonance aspects of the C-Se position, as well as the binding of the pendant N-group; the coumarin lactone carbonyl also allows for resonance to be achieved (vide infra). The leaving group, -SePh, while precedented in some systems, depends on electronic tuning (Fig. 1). For 1, the response times with GSH was ~100ms; a 100-fold fluorescence increase is observed (Compound 1). The probe also reacts with cysteine (Cys) and homocysteine (Hcy), albeit differently. For glutathione probing, the greater wavelength maxima (1: 550nm, DACP-1: 555nm, DACP-2: 590nm) enabled eventual cell studies (confocal microscopy) and animal studies. The limits of detection (LOD, 1: 270nM DACP-1: 10.1nM DACP-2: 17.0nM), as measured using the 3σ/k method. We provide a didactic presentation from probe conception to probe in vivo testing, etc., with additional considerations presented; a variety of factors/issues (2.1-2.28) help maintain a realistic sequence, a flow from wider to narrower, of the factors that go into developing medical, biological and neurodegenerative disease-related probes, meant to help other researchers follow our intention, gain perspective, and overcome current limitations.
Assuntos
Doenças Neurodegenerativas , Selênio , Aldeídos , Animais , Cumarínicos , Cisteína , Corantes Fluorescentes , Glutationa , Células HeLa , HumanosRESUMO
Advancements in metal nanoparticle synthesis using plant extracts and their anticancer activity have received significant attention in recent years. The green approach for the synthesis of gold nanoparticles (AuNPs) using leaf extract of Sasa borealis is reported in this study. Synthesis of AuNPs was performed at 50 °C, and nanoparticle formation was observed after 20 min incubation. AuNPs formation was confirmed by the UV-visible spectrum peak at 542 nm. The synthesized AuNPs were oval, spherical with sizes around 10-30 nm observed using the transmission electron microscope. Energy dispersive X-ray analysis was utilized for the detection of elemental compound. The face centered cubic structure was confirmed by X-ray diffraction pattern. The reduction of tetrachloroauric acid into AuNPs by the phytochemical compounds of S. borealis extract was determined by Fourier transform infrared spectroscopy and the presence of biomolecules was studied by GC-MS. The synthesized AuNPs was tested for toxic effect on HEK293 cells and anticancer activity on AGS cells by WST-1® assay. Condensation or fragmentation is a characteristics of apoptosis, which was confirmed by 4,6-diamidino-2-pheynylindole dihydrochoride (DAPI) staining. The S. borealis-mediated AuNPs have good activity as an anticancer agent and it will be beneficial in cancer therapeutics.