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Nature Communications Jun 2024Co-immobilization of cells and enzymes is often essential for the cascade biocatalytic processes of industrial-scale feasibility but remains a vast challenge. Herein, we...
Co-immobilization of cells and enzymes is often essential for the cascade biocatalytic processes of industrial-scale feasibility but remains a vast challenge. Herein, we create a facile co-immobilization platform integrating enzymes and cells in covalent organic frameworks (COFs) to realize the highly efficient cascade of inulinase and E. coli for bioconversion of natural products. Enzymes can be uniformly immobilized in the COF armor, which coats on the cell surface to produce cascade biocatalysts with high efficiency, stability and recyclability. Furthermore, this one-pot in situ synthesis process facilitates a gram-scale fabrication of enzyme-cell biocatalysts, which can generate a continuous-flow device conversing inulin to D-allulose, achieving space-time yield of 161.28 g L d and high stability (remaining >90% initial catalytic efficiency after 7 days of continuous reaction). The created platform is applied for various cells (e.g., E. coli, Yeast) and enzymes, demonstrating excellent universality. This study paves a pathway to break the bottleneck of extra- and intracellular catalysis, creates a high-performance and customizable platform for enzyme-cell cascade biomanufacturing, and expands the scope of biocatalysis process intensification.
Topics: Biocatalysis; Escherichia coli; Enzymes, Immobilized; Glycoside Hydrolases; Cells, Immobilized; Metal-Organic Frameworks; Saccharomyces cerevisiae
PubMed: 38951487
DOI: 10.1038/s41467-024-49831-8 -
Journal of the American Chemical Society Jul 2024Here, we report a set of new polymerization reactions enabled by the 1,2-regioselective hydro- and silylcupration of enyne-type propargylic electrophiles. Highly...
Here, we report a set of new polymerization reactions enabled by the 1,2-regioselective hydro- and silylcupration of enyne-type propargylic electrophiles. Highly regioregular head-to-tail poly(2-butyne-1,4-diyl)s (HT-PBD), bearing either methyl or silylmethyl side chains, are synthesized for the first time. A rapid entry into carbon-rich copolymers with adjustable silicon content is developed via in situ monomer bifurcation. Furthermore, a one-pot polymerization/semireduction sequence is developed to access a -poly(butadiene)-derived backbone by a ligand swap on copper hydride species. Interestingly, borocupration, typically exhibiting identical regioselectivity with its hydro- and silyl analogues, seems to proceed in a 3,4-selective manner. Computational studies suggest the possible role of the propargylic leaving group in this selectivity switch. This work presents a new class of regioregular -carbon-rich polymers and meanwhile a novel approach to organosilicon materials.
PubMed: 38951483
DOI: 10.1021/jacs.4c05524 -
Methods in Molecular Biology (Clifton,... 2024Nonviral transfection has been used to express various recombinant proteins, therapeutics, and virus-like particles (VLP) in mammalian and insect cells. Virus-free...
Nonviral transfection has been used to express various recombinant proteins, therapeutics, and virus-like particles (VLP) in mammalian and insect cells. Virus-free methods for protein expression require fewer steps for obtaining protein expression by eliminating virus amplification and measuring the infectivity of the virus. The nonviral method uses a nonlytic plasmid to transfect the gene of interest into the insect cells instead of using baculovirus, a lytic system. In this chapter, we describe one of the transfection methods, which uses polyethyleneimine (PEI) as a DNA delivery material into the insect cells to express the recombinant protein in both adherent and suspension cells.
Topics: Animals; Recombinant Proteins; Transfection; Polyethyleneimine; Plasmids; Insecta; Sf9 Cells; Cell Line; Gene Expression; Spodoptera
PubMed: 38951345
DOI: 10.1007/978-1-0716-3961-0_23 -
Environmental Monitoring and Assessment Jun 2024Cigarette butts (CBs) and Microplastics (MPs) have serious harmful effects on the environment and living organisms despite their small size. This research aims to...
Cigarette butts (CBs) and Microplastics (MPs) have serious harmful effects on the environment and living organisms despite their small size. This research aims to investigate the abundance and pollution status of CBs and MPs in Tahtalı Dam Basin (West Anatolia, Türkiye) which is the most important drinking and irrigation water resources. Clean Environment Index (CEI) and Cigarette Butt Pollution Index (CBPI) were used to determine pollution degree of the basin. The total number of CBs were 1.478 items, the total number of MPs were 477 items/m in the basin. As a result of this study, MP particles weren't found in Balaban Stream. Highest number of MP particles observed in 100-250 μm (45%) size class. The most abundant MP type and colour were, fragment (54%) and white (42%), respectively. Polyethylene terephthalate (50%) was the most abundant type of polymer according to the ATR- FTIR analysis. As a result of the CEI and CBPI, the upstream stations of the stream were classified as "clean" status, while downstream sampling points of the stream and Balaban Lake coasts were classified as "extremely dirty" status. The calculated volumes of MP particulates from mining facility, agricultural and recreational activities indicate that anthropogenic factors are the most important MP source in the Tahtalı Dam Basin. This study is the first study about MP and CB pollution of the freshwater ecosystems in the region.
Topics: Turkey; Water Pollutants, Chemical; Environmental Monitoring; Microplastics; Risk Assessment; Drinking Water; Agricultural Irrigation; Rivers
PubMed: 38951278
DOI: 10.1007/s10661-024-12814-9 -
Environmental Monitoring and Assessment Jul 2024Microplastics, an emerging contaminant, are widespread in oceans around the world, and rivers are the key conveyors of these pollutants into the oceans. There exists a...
Microplastics, an emerging contaminant, are widespread in oceans around the world, and rivers are the key conveyors of these pollutants into the oceans. There exists a dearth of available data pertaining to seasonal fluctuation, spatial distribution and risk assessment of microplastics in rivers extending from upper reaches to the lower reaches. The collection of such data is of utmost importance for the purpose of formulating beneficial management strategies for riverine microplastics. In order to bridge this research gap, an investigation was made in the Periyar River in Kerala, India, which is exposed to anthropogenic stress and is at risk of microplastic pollution. A total of eighteen sites (six sites each from downstream, midstream and upstream) along the 244 km of the river were investigated across three seasons in a year. The study revealed a discernible pattern in the spatial distribution of microplastic concentrations, wherein there was a rise in abundance from the upstream to midstream and then a sudden increase of abundance along the downstream regions towards the lower reaches. The highest mean microplastic abundance of 124.95 items/L was obtained during the monsoon season followed by post-monsoon season i.e. 123.21 items/L and pre-monsoon i.e. 120.50 items/L. The predominant forms of microplastics were found to be fibres, fragments and filaments. Most prevalent polymer types acquired were polyethylene (PE) and polypropylene (PP). Pollution hazard index (PHI) and pollution load index (PLI) were also evaluated to assess the water quality of this river. The findings of this study conclude that the Periyar River is polluted with microplastics throughout its course and offer significant insights into the detection of microplastic origins in river systems and lend support to the implementation of potential measures aimed at mitigating their impact.
Topics: India; Water Pollutants, Chemical; Rivers; Environmental Monitoring; Microplastics; Risk Assessment; Seasons; Water Pollution, Chemical
PubMed: 38951273
DOI: 10.1007/s10661-024-12820-x -
Mikrochimica Acta Jun 2024Cystinuria is a genetic disorder, and in severe cases, it might lead to kidney failure. As an important biomarker for cystinuria, the level of arginine (Arg) in urine is...
Cystinuria is a genetic disorder, and in severe cases, it might lead to kidney failure. As an important biomarker for cystinuria, the level of arginine (Arg) in urine is a vital indicator for cystinuria screening. Therefore, it is urgently needed to detect Arg with high selectivity and sensitivity. In this work, a boric acid functionalized Zr-based metal-organic framework UiO-PhbA is prepared by grafting phenylboronic acid on UiO-66-NH through a Schiff base reaction using a covalent post-synthesis modification (CPSM) strategy. The prepared UiO-PhbA exhibits a sensitive and specific fluorescence "turn-on" response to Arg and can be exploited to detect Arg in human serum and urine samples with a broad linear range of 0.6-350 µM and low limit of detection (LOD) of 18.45 nM. This study provides a new and reliable rapid screening protocol for sulfite oxidase deficiency-related diseases.
Topics: Humans; Cystinuria; Metal-Organic Frameworks; Fluorescent Dyes; Arginine; Limit of Detection; Biomarkers; Boronic Acids; Spectrometry, Fluorescence; Zirconium
PubMed: 38951266
DOI: 10.1007/s00604-024-06519-6 -
Planta Jul 2024Our findings shed light on the regulation of anthocyanin and proanthocyanidin biosynthesis in chickpea seed coats. Expression of R2R3-MYB transcription factors CaLAP1...
Our findings shed light on the regulation of anthocyanin and proanthocyanidin biosynthesis in chickpea seed coats. Expression of R2R3-MYB transcription factors CaLAP1 and CaLAP2 enhanced the anthocyanins and proanthocyanidins content in chickpea. The seed coat color is a major economic trait in leguminous crop chickpea (Cicer arietinum). Anthocyanins and proanthocyanidins (PAs) are two classes of flavonoids that mainly contribute to the flower, seed coat and color of Desi chickpea cultivars. Throughout the land plant lineage, the accumulation of anthocyanins and PAs is regulated by MYB and bHLH transcription factors (TFs), which form an MBW (MYB, bHLH, and WD40) complex. Here, we report two R2R3-MYB TFs in chickpea belonging to the anthocyanin-specific subgroup-6, CaLAP1 (Legume Anthocyanin Production 1), and CaLAP2 (Legume Anthocyanin Production 2), which are mainly expressed in the flowers and developmental stages of the seeds. CaLAP1 and CaLAP2 interact with TT8-like CabHLH1 and WD40, forming the MBW complex, and bind to the promoter sequences of anthocyanin- and PA biosynthetic genes CaCHS6, CaDFR2, CaANS, and CaANR, leading to anthocyanins and PA accumulation in the seed coat of chickpea. Moreover, these CaLAPs partially complement the anthocyanin-deficient phenotype in the Arabidopsis thaliana sextuple mutant seedlings. Overexpression of CaLAPs in chickpea resulted in significantly higher expression of anthocyanin and PA biosynthetic genes leading to a darker seed coat color with higher accumulation of anthocyanin and PA. Our findings show that CaLAPs positively modulate anthocyanin and PA content in seed coats, which might influence plant development and resistance to various biotic and abiotic stresses.
Topics: Cicer; Seeds; Anthocyanins; Plant Proteins; Gene Expression Regulation, Plant; Proanthocyanidins; Transcription Factors; Plants, Genetically Modified; Arabidopsis; Flowers
PubMed: 38951258
DOI: 10.1007/s00425-024-04470-7 -
Nature Biomedical Engineering Jul 2024By combining living cells with therapeutics, cell-drug conjugates can potentiate the functions of both components, particularly for applications in drug delivery and... (Review)
Review
By combining living cells with therapeutics, cell-drug conjugates can potentiate the functions of both components, particularly for applications in drug delivery and therapy. The conjugates can be designed to persist in the bloodstream, undergo chemotaxis, evade surveillance by the immune system, proliferate, or maintain or transform their cellular phenotypes. In this Review, we discuss strategies for the design of cell-drug conjugates with specific functions, the techniques for their preparation, and their applications in the treatment of cancers, autoimmune diseases and other pathologies. We also discuss the translational challenges and opportunities of this class of drug-delivery systems and therapeutics.
PubMed: 38951139
DOI: 10.1038/s41551-024-01230-6 -
Scientific Reports Jul 2024Polyethylene glycols (PEGs) are used in industrial, medical, health care, and personal care applications. The cycling and disposal of synthetic polymers like PEGs pose...
Polyethylene glycols (PEGs) are used in industrial, medical, health care, and personal care applications. The cycling and disposal of synthetic polymers like PEGs pose significant environmental concerns. Detecting and monitoring PEGs in the real world calls for immediate attention. This study unveils the efficacy of time-of-flight secondary ion mass spectrometry (ToF-SIMS) as a reliable approach for precise analysis and identification of reference PEGs and PEGs used in cosmetic products. By comparing SIMS spectra, we show remarkable sensitivity in pinpointing distinctive ion peaks inherent to various PEG compounds. Moreover, the employment of principal component analysis effectively discriminates compositions among different samples. Notably, the application of SIMS two-dimensional image analysis visually portrays the spatial distribution of various PEGs as reference materials. The same is observed in authentic cosmetic products. The application of ToF-SIMS underscores its potential in distinguishing PEGs within intricate environmental context. ToF-SIMS provides an effective solution to studying emerging environmental challenges, offering straightforward sample preparation and superior detection of synthetic organics in mass spectral analysis. These features show that SIMS can serve as a promising alternative for evaluation and assessment of PEGs in terms of the source, emission, and transport of anthropogenic organics.
Topics: Cosmetics; Spectrometry, Mass, Secondary Ion; Polyethylene Glycols; Principal Component Analysis
PubMed: 38951137
DOI: 10.1038/s41598-024-65504-4 -
Acta Biomaterialia Jun 2024The precise mechanisms underlying the cellular response to static electric cues remain unclear, limiting the design and development of biomaterials that utilize this...
The precise mechanisms underlying the cellular response to static electric cues remain unclear, limiting the design and development of biomaterials that utilize this parameter to enhance specific biological behaviours. To gather information on this matter we have explored the interaction of collagen type-I, the most abundant mammalian extracellular protein, with polyvinylidene fluoride (PVDF), an electroactive polymer with great potential for tissue engineering applications. Our results reveal significant differences in collagen affinity, conformation, and interaction strength depending on the electric charge of the PVDF surface, which subsequently affects the behaviour of mesenchymal stem cells seeded on them. These findings highlight the importance of surface charge in the establishment of the material-protein interface and ultimately in the biological response to the material. STATEMENT OF SIGNIFICANCE: The development of new tissue engineering strategies relies heavily on the understanding of how biomaterials interact with biological tissues. Although several factors drive this process and their driving principles have been identified, the relevance and mechanism by which the surface potential influences cell behaviour is still unknown. In our study, we investigate the interaction between collagen, the most abundant component of the extracellular matrix, and poly(vinylidenefluoride) with varying surface charges. Our findings reveal substantial variations in the binding forces, structure and adhesion of collagen on the different surfaces, which collectively explain the differential cellular responses. By exposing these differences, our research fills a critical knowledge gap and paves the way for innovations in material design for advanced tissue regeneration strategies.
PubMed: 38950807
DOI: 10.1016/j.actbio.2024.06.039