Extracellular vesicle (EV) research and applications increasingly leverages porous membrane technologies beyond traditional filtration methods used for isolation and purification. While filtration remains foundational in EV handling, emerging applications employ membranes directly in new EV research contexts, such as: three-dimensional cell culture, tissue barrier modeling, environmental monitoring, and horizontal EV transfer studies. Concurrently, advancements in membrane science are providing novel modifications leading to significantly enhanced membrane performance in EV applications. These innovations include: surface-enhanced membranes for plasmonic detection, selective EV corona characterization and manipulation, targeted EV removal strategies, self-assembled and patterned membranes to simplify production and improve consistency, customized pore geometries for optimized throughput, and self-cleaning membranes to mitigate fouling. Collectively, developments in these porous membrane technologies promise to markedly improve the effectiveness, precision, and scope of EV-based research and applications, particularly within diagnostics, therapeutics, and cancer biology.
Microbial biosurfactants are emerging as sustainable alternatives to synthetic surfactants due to their biodegradability, low toxicity, and multifunctional biological activities. Their potential in biomedical and pharmaceutical applications is increasingly recognized, yet integrated evaluations that link production strategies to translational outcomes remain limited. This review critically examines recent advances in biosurfactant production, optimization, and purification, as well as their applications in drug delivery, nanomedicine, antimicrobial therapy, vaccine formulation, wound healing, and immunomodulation. Major classes, including glycolipids, lipopeptides, phospholipids, and polymeric biosurfactants, are compared in terms of physicochemical properties and therapeutic potential. Evidence shows biosurfactants can reduce bacterial adhesion by up to 90%, inhibit biofilm formation by over 80%, and enhance antibiotic efficacy against multidrug-resistant pathogens. Biosurfactant-based carriers such as liposomes, micelles, nanoemulsions, and nanoparticles improve drug solubility, stability, bioavailability, and targeted release. Advances in engineered microbial platforms, low-cost substrates, and process optimization have enhanced production feasibility, though clinical translation remains constrained by cost, downstream processing, formulation stability, safety, and regulatory hurdles. Emerging approaches, including AI-assisted optimization and multiomics-guided discovery, promise next-generation biosurfactants with superior therapeutic performance.
Rare sugars have attracted substantial interest owing to their critical roles in agriculture, cosmetics, food, and pharmaceutical industries. Traditional chemical synthesis methods face limitations such as low selectivity and environmental concerns. Consequently, enzymatic synthesis of rare sugars has emerged as an increasingly prominent research focus. To address the limitations of single-enzyme systems, multi-enzyme reactions have been developed as promising approaches, encompassing isomerases, epimerases, aldolases, phosphatases, oxidases, dehydratases, and associated coenzymes. Through the utilization of multi-enzyme systems, substrate costs can be reduced while the overall yields of rare sugars are enhanced. Furthermore, the application of immobilized enzymes is critical for optimizing the production process. Immobilization techniques can reduce reaction costs, improve enzyme properties, and enhance efficiency and stability. This review summarizes multi-enzyme systems for rare sugars biosynthesis and diverse immobilization methods, including traditional approaches, innovative nanomaterial-based techniques, and the use of scaffold proteins. Additionally, this review incorporates a techno-economic analysis of rare sugars production, addressing bottlenecks associated with enzyme stability and process costs, aiming to provide guidance for the industrial translation of rare sugar manufacturing technologies.
Antibiotic resistance is an emerging global issue that has reduced the efficacy of antibiotics for treating life-threatening bacterial infections. Bacterial adaptive enzymatic defense mechanisms allow cells to activate or modify specific enzymes that inactivate antibiotics and support survival under antimicrobial stress. Antibiotics are predominantly inactivated through enzymatic degradation or chemical modification. Most of the β-lactamases, macrolide esterases, tetracycline-modifying enzymes, fosfomycin degrading enzymes, aminoglycoside-modifying enzymes, and other bacterial enzymes chemically modify or degrade the antibiotics making them inactive. This review covers classification and mechanisms of enzyme-mediated resistance and emphasizes the significant enzymes involved in inactivation of various antibiotic classes. It also summarizes recent biotechnological advances to combat antibiotic resistance, including β-lactamase inhibitors, phage therapy, antimicrobial peptides, and CRISPR-Cas9 systems, along with emerging therapeutic approaches and current trends in antibiotic research. A deeper understanding of enzyme-mediated resistance and the cellular intelligence driving bacterial adaptation is crucial for designing effective therapeutic strategies, preserving antibiotic efficacy, and reducing the global burden of resistant infections.
The production of second-generation bioethanol from lignocellulosic biomass is a promising solution for sustainable energy, yet it faces significant challenges also due to the inhibitory effects of weak acids released during biomass pretreatment, particularly acetic, formic and levulinic acids. This review describes the ability of Saccharomyces cerevisiae, with a focus on natural isolates, in overcoming these challenging compounds. Indeed, natural isolates exhibit greater genetic and phenotypic diversity than laboratory and industrial strains, offering unique traits such as enhanced stress tolerance, metabolic efficiency, and adaptive responses to weak acids. This investigation explores the transcriptional and genomic mechanisms underlying yeast adaptive responses, emphasizing key regulatory networks and resistance pathways, including drug H+ antiporters, Reactive Oxygen Species (ROS) mitigation strategies, and membrane composition adjustments. Strategies for strains improvement, involving adaptive laboratory evolution (ALE), genome shuffling, and hybridization, are also discussed as complementary approaches to develop robust yeast capable of thriving under stressful industrial fermentation conditions. The integration of these techniques, along with genomic and transcriptomic insights, provides a comprehensive framework for engineering high-performance yeast strains. Ultimately, this review underscores the potential of leveraging natural diversity and innovative biotechnological strategies to advance the scalability and efficiency of lignocellulosic bioethanol production through S. cerevisiae fermentation.
With the continuing improvement of living standards, humans have been paying more attention to the influence of diet on their health and disease prevention. One group of the most important and essential foods are those rich in polyunsaturated fatty acids (PUFAs), in particular, ω-3 PUFAs. Beyond basic nutritional function, ω-3 PUFAs have been shown to have considerable beneficial effects on human growth and development as well as for the prevention and treatment of nonalcoholic fatty liver, autoimmune reactions, and various chronic illnesses. Although ω-3 PUFAs can be found in some terrestrial organisms, marine microorganisms, including bacteria, fungus-like organisms, and microalgae serve, as the richest natural source of ω-3 PUFAs. Through the food chain, ω-3 PUFAs can also be enriched in marine and freshwater organisms that are commonly consumed as part of the human diet (fish, crustaceans, mollusks, etc.). As the food consumption of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), two representative essential ω-3 PUFAs, continues to grow, it is becoming increasingly crucial to develop efficient and sustainable systems toward production of these valuable fatty acids. This article summarizes the recent advancements in the biotechnological production of PUFAs, especially EPA and DHA, including strategies adopted to increase the production of ω-3 PUFAs and the development of some new microbial strains or plants able to produce DHA and EPA.
Early detection of breast cancer is crucial for improving women's health and survival rates. However, traditional diagnostic and treatment approaches are often expensive, time-intensive, complex, and may lack the sensitivity and specificity needed for effective clinical use. Detecting early-stage cancer and monitoring recurrence demand highly sensitive, reproducible detection of low-abundance biomarkers. Electrochemical biosensors offer a promising solution, enabling rapid, cost-effective diagnostics suitable for point-of-care (POC) applications. Among these, carbon-based nanomaterials, such as: carbon nanotubes (CNTs), graphene, graphene oxide (GO), reduced graphene oxide (rGO), carbon dots (CDs), and their nanocomposites, stand out for their unique properties that support the sensitive detection of breast cancer biomarkers. This review comprehensively explores recent advancements (2020 to March 2025) in using carbon-based materials for POC electrochemical sensors in breast cancer diagnostics. It covers synthesis methods, structural characteristics, and detection mechanisms associated with key biomarkers, like: HER2, CA 15-3, CEA, CA 125, BRCA1, and MUC1, including performance metrics such as linear range, limit of detection (LOD), and sensitivity. Additionally, the potential integration of these biosensors with smartphones is discussed, along with an overview of sustainability challenges and future research directions in this field.
The persistent environmental contamination resulting from per- and polyfluoroalkyl substances (PFAS) presents substantial challenges to both environment and public health, thereby underscoring the persistent necessity for sustainable remediation approaches. The removal of PFAS from contaminated water using sorbent materials, particularly sustainable and green materials, is a critical area of research due to their inherent biodegradability, biocompatibility, and eco-friendly properties. This review explores recent progress in sustainable and eco-friendly strategies aimed at PFAS immobilization through the creation of novel sorbent materials. By emphasizing the incorporation of biodegradable and nontoxic constituents, we elucidated the improved adsorption efficacy of engineered sorbents for the effective sequestering of PFAS compounds. A thorough examination of various batch experiments indicated that these novel sorbents exhibit enhanced PFAS removal efficiency compared to conventional remediation techniques. Moreover, we highlight the challenges that remain despite the promising attributes of sustainable and eco-friendly sorbents for PFAS removal and deliberate on prospective opportunities for their extensive utilization in the ongoing efforts to combat PFAS pollution.
Biomanufacturing advances sustainable production and circular economy, while wastes-derived one-carbon (C1) compounds (e.g., CO2, methanol, formate and methane) expand substrate diversity, accelerate wastes valorization, and mitigate climate change. The design of C1 feedstocks-driven biorefineries is often hindered by: limited genetic toolkits, impaired cell growth, inefficient substrate utilization, and unclear metabolic mechanism. In this context, this review addresses these challenges through comprehensive analysis of: pathway exploitation, metabolic regulation, emerging technology, and biochemicals synthesis for optimizing C1-trophic performance. We first analyze key bottlenecks in enhancing assimilation efficiency of natural C1-utilizers, and then systematically summarize the strategies for harnessing nontraditional feedstocks using: engineered autotrophs, methylotrophs, formatotrophs, and methanotrophs. Importantly, we outline a bottom-up framework on systematic and modular redesign of C1-driven microbial cell factories for promoting industrial applications. Finally, we identify unresolved challenges and strategic opportunities to guide environmental preservation and performance optimization. Overall, this review provides a roadmap for transformative progress in sustainable biomanufacturing and wastes re-utilization.
Harnessing biomass for bio-based industrial biotechnology is vital for addressing global energy needs and mitigating climate change. In this context, microorganisms are the cornerstone of biorefineries based on renewable materials, with applications in bioenergy, agriculture, biomedicine, and other sectors. By engineering metabolic pathways, microorganisms can be tailored to improve yields, tolerate industrial conditions, and selectively produce valuable compounds. Through advances in metabolic engineering and synthetic biology, engineered strains of the yeast Saccharomyces cerevisiae have been successfully developed to efficiently convert the pentose sugars D-xylose and L-arabinose. Despite this important breakthrough, the efficient transport of these sugars remains a major limitation. Sugar sensing and transport in yeast are regulated at both transcriptional and post-translational levels. D-xylose is not recognized as a fermentable carbon source, leading to downregulation of transporter expression, removal from the cytoplasmic membrane, and degradation via ubiquitination in the absence of extracellular glucose. Additionally, transporters exhibit lower affinity for C5 sugars compared to D-glucose, resulting in strong D-glucose repression. To address these challenges, cutting-edge strategies have been successfully employed, including rational protein engineering, directed evolution, and machine learning approaches, to expand the repertoire of C5 transporters available for engineering in S. cerevisiae. Specific D-xylose transporters have been redesigned, with key residues identified to reduce D-glucose affinity, while studies have demonstrated improvements in transporter stability and sugar uptake rates. This review summarizes the key bottlenecks in C5 sugar transport and highlights the major advances and progress made toward creating robust microbial platforms capable of sustainable and efficient bio-based production.
Cold stress (CS) is a major abiotic factor that significantly affects rice growth, development, and productivity. To withstand CS, rice plants have evolved intricate mechanisms that enable them to detect external signals and respond to changing environmental conditions. Understanding the physiological and molecular mechanisms underlying cold tolerance (CT) is therefore crucial for developing rice cultivars that can thrive under cold conditions. To date, a limited number of genes/QTLs associated with CT, such as: Ctb1, COLD1, COLD6, qLTG3-1, SGD1, and CTB4a, have been cloned and well studied. These genes play essential roles in the plant's response to CS by regulating various stress-related pathways, including reactive oxygen species (ROS) scavenging, osmotic regulation, and signal transduction. The physical co-localization of OsSRFP1 (associated with CT at the seedling stage) and OsMTACP2 (associated with CT at the reproductive stage) on chromosome 3 (between 13.1-13.3 Mb) suggests the potential for simultaneous introgression of these traits into recipient cultivars using donor parents carrying the desirable alleles. Genomic selection, powered by high-throughput genotyping technologies, enhances the accuracy and efficiency of identifying cold-tolerant genotypes. Additionally, integrating multi-omics approaches with gene-editing technologies offers a comprehensive strategy to uncover the complex molecular networks and regulatory pathways involved in CT, thereby identifying potential targets for genetic improvement. Overall, this review emphasizes the potential of integrating molecular, physiological, and genomic tools to develop rice cultivars resilient to CS, thereby contributing to global food security in the context of climate change.
The production and supply of quality food to the ever-growing global population is becoming a foremost challenge due to ever-increasing losses in crops yield. The yield losses in crops are majorly due to various biotic factors, such as: microbial plant pathogens, insect pests, weeds, etc. Therefore, there is an urgent need to develop modern agricultural practices that can effectively minimize losses and enhance agricultural production and yield. To achieve this target several synthetic agrochemicals, such as fertilizers, insecticides, pesticides, etc., are being extensively used in the current agricultural practices. However, it is well-known that uncontrolled and excessive usage of synthetic agrochemicals directly or indirectly affects the ecosystem and causes numerous undesirable effects. Therefore, to overcome these problems, the application of biopesticides is recommended as one of the eco-friendly and sustainable approaches. In this context, the application of biopesticides like Bacillus thuringiensis (Bt) in agriculture is increasing day by day. However, despite better pesticidal efficacy, widespread use of Bt-based biopesticides is limited because of their degradation after exposure to UV radiation. Hence, the development of Bt biopesticides resistant to UV radiation is extremely necessary. In this review, we discuss the role of Bt biopesticides in the management of plant insect pests. Moreover, various limitations in the use of conventional UV protectants in Bt are also highlighted. Similarly, the emerging role of nanotechnology in protecting Bt biopesticide from UV radiation is discussed in depth.
Metabolic diseases, such as obesity and diabetes, have risen due to lifestyle changes. Traditional treatments, including dietary modifications and pharmacological interventions, are limited by low compliance and adverse effects, highlighting the need for alternative therapeutic approaches that offer improved patient compliance and long-term effectiveness. Engineered live biotherapeutic products (eLBPs) have emerged as a promising strategy that combines bacterial chassis with synthetic genetic circuits for precise and targeted disease treatment. Unlike conventional therapeutics, eLBPs can colonize the intestinal tract and enable localized and condition-responsive therapeutic activity while offering improved safety profiles through defined mechanisms of action. This review highlights key strategies for eLBP development, particularly chassis selection and genetic circuit design. Applications in metabolic diseases, including inherited disorders such as phenylketonuria (PKU), demonstrate how engineered gene circuits can modulate specific metabolic pathways. However, several challenges remain, including genetic stability, interindividual variability, biological safety, and production scalability. In addition, further research on host-microbiota interactions is required to improve therapeutic predictability and efficacy, supporting the development of safe and effective personalized eLBP-based therapies for metabolic diseases.
Surface-enhanced Raman spectroscopy (SERS) is rapidly emerging as a transformative technology in dermatological diagnostics, offering ultra-sensitive, noninvasive detection of molecular markers associated with skin diseases. With nearly five billion individuals affected worldwide and conventional diagnostic methods often limited by invasiveness or subjective interpretation, there is an urgent need for rapid, accessible, and real-time diagnostic solutions. The present review systematically examines the integration of SERS into dermatological practice, with a focus on recent advancements in biosensing platforms, nanostructure engineering, and point-of-care devices. Innovative methodologies, including microneedle-based SERS biosensors and microfluidic-integrated detection systems, are discussed in the context of their ability to enhance diagnostic accuracy for early-stage skin cancers, microbial infections, and inflammatory dermatoses. Furthermore, the review highlighted the role of AI-driven spectral analysis in improving data interpretability and clinical decision-making. Critical evaluation of the challenges of substrate reproducibility, clinical standardization, and device scalability, while outlining emerging strategies that aim to bridge laboratory innovations with clinical applications, are explored. Looking ahead, the development of portable, low-cost SERS platforms for continuous skin health monitoring, combined with personalized diagnostic pathways, is poised to redefine dermatological care and expand the scope of precision medicine.
L-Asparaginase (E.C.3.5.1.1; L-ASNase) hydrolyzes L-asparagine (an essential amino acid for the growth of leukemic cells) to aspartic acid and ammonia. It is obtained from various sources, including: bacteria, yeast, fungi, plants, and animals. It is used as a chemotherapeutic agent to treat acute lymphoblastic leukemia (ALL) and reduce acrylamide formation in baked and fried foods. Globally, various recombinant and pegylated L-ASNases formulations, including Escherichia coli and Erwinia-derived variants, are in Phase II or active clinical trials for ALL and related conditions across the USA, Brazil, and Canada. Some drugs are recruiting, and other variants of E. coli and E. chrysanthemi L-ASNases are already approved for marketing. Although L-ASNase is used as a therapeutic agent, immunogenic reactions and other adverse effects continue to limit its use. To enhance yield, L-ASNase isoforms are cloned and expressed in host cells, generating recombinants with distinct physicochemical properties and kinetic parameters. The enzymes' temperature and pH ranges vary from 25 to 100 °C and 6 to 10, respectively. Nowadays, enzymatic modifications, such as immobilization (chemical, physical, and PEGylation), mutagenesis, and PASylation, are used to overcome the limitations of commercialized L-ASNase-based drugs. Therefore, this review is a timely effort to compile and analyze the properties of recombinant L-ASNases and the contemporary techniques used to improve L-ASNase. A comprehensive study would help us better understand the kinetic parameters, biochemical properties, and modification trends of L-ASNase, enabling the development of robust, reliable therapeutics in the future.
Poly-γ-glutamic acid (γ-PGA) is a natural biopolymer with broad application potential. Molecular weight (MW) is a key physicochemical parameter governing its structural properties, functional performance, and application scope. Recently, sustainable biosynthesis of γ-PGA with controllable MW has gained attention because of its environmental sustainability and process flexibility. With advances in molecular editing and synthetic regulation, MW-control strategies have shifted from random mutagenesis-based strain improvement to precise gene-engineering regulation. This review surveys natural microbial resources producing γ-PGA with diverse MWs and engineering strategies enabling de novo γ-PGA biosynthesis in multiple microbial chassis. To address limited production efficiency, we summarize metabolic engineering approaches for improving γ-PGA yield and MW tunability, including precursor supply optimization, carbon-flux redistribution, transcriptional regulation, use of non-food renewable substrates, and mitigation of metabolic burden from multi-layered engineering. To promote customized production of low-MW γ-PGA, we highlight hydrolase-centered strategies, emphasizing hydrolase screening, optimization of expression elements, and coordinated regulation of γ-PGA stereochemical configuration. Finally, we review applications of γ-PGA with different MWs in food, biopharmaceutical, and agricultural sectors, critically examine links between molecular characteristics and application requirements, and discuss future functional diversification and industrial-scale development.
Alcohol dehydrogenases (ADHs) are widely used to prepare chiral alcohols because of their environmental benefits, high-cost effectiveness, broad substrate acceptance, and high enantioselectivity. However, the stereoselective reduction of "difficult-to-reduce" ketones, which are characterized by bulky substituents, similar steric and electronic properties at the α, α'-positions, or complex structures, remains a significant challenge. Understanding the interaction between these ketones and enzymes is crucial for overcoming catalytic limitations. This review summarized recent research advances in the asymmetric reduction of "difficult-to-reduce" ketones. We specifically discuss the stereoselective recognition mechanisms and outline strategies for regulating enzyme selectivity based on enzyme-substrate interactions. Finally, we propose future perspectives on the rational design of ADHs for the synthesis of high-value chiral chemicals.
Endophytic microorganisms are a vital part of the plant microbiome, contributing significantly to the plant's growth, development, and stress tolerance. Proteomics investigations have significantly enhanced our comprehension of the interactions between plants and endophytes, illuminating the complex molecular mechanisms that govern these mutually beneficial relationships. The review aims to integrate the latest developments in proteomic research concerning endophyte-plant interactions, emphasizing on elucidating the molecular mechanisms that underlie the benefit imparted to the host plant by the symbionts. The special focus of the review is to discuss the proteome level changes happening at the early recognition events, primary and secondary metabolism, signaling pathways, and defense mechanisms. By underscoring critical proteomic signatures, the review aspires to offer insights into how these interactions enhance plant health, increase stress resilience, and promote overall growth. The article discusses the potential applications of proteomics in agriculture and environmental sciences, emphasizing its role in crop resilience against biotic and abiotic stresses, optimizing biocontrol strategies, and improving nutrient use efficiency. The article also highlights that despite the advancements, critical gaps persist including the necessity for a deeper understanding of the temporal dynamics of proteomic responses, the specificity of protein-protein interactions, and the influence of environmental factors on the proteome induced by the endophytes. The review concludes by proposing future directions for proteomics research in plant-endophyte interactions for developing a more comprehensive understanding of the intricate molecular dialogues for developing a more sustainable and resilient agricultural systems.
With the global population projected to reach 9.8 billion by 2050, coupled with extreme weather events and resource scarcity, there is an urgent need to redesign existing Green Revolution varieties (GRVs) for a modern revolution in agriculture. Brassinosteroids (BRs) play crucial roles in shaping plant architecture and hold significant potential for developing modern GRVs that are high-yielding, climate-resilient and environmentally sustainable. Recent advances in crop improvement, particularly by manipulating BR signaling and biosynthesis genes, have already generated semi-dwarf GRVs. These BR-modified GRVs exhibited enhanced overall crop performance, including yield, photosynthetic capacity and nitrogen-use efficiency. However, they still face challenges regarding tolerance to various biotic and abiotic stresses, although studies on BR mutants have indicated that BRs can confer resilience to biotic and abiotic stresses. Despite such studies having made significant progress in exploring the role of BRs, their functions in crop species are not comprehensively understood. Therefore, additional BR mutants need to be developed to make existing crops climate-resilient and to tailor them to specific regional needs, such as increased frost tolerance in Eastern Europe. Ongoing efforts are focused on identifying candidate BR-related genes in crops to reveal the regulatory mechanisms underlying BR signaling and biosynthesis, thereby improving overall performance. This review highlights recent advances in BR research related to crop improvement and stress adaptation. Moreover, it emphasizes the potential of BRs to drive a new generation of BR-modified GRVs and outlines strategies to achieve this by manipulating BR signaling and biosynthesis pathways to develop high-yield, climate-resilient crops.
Mass spectrometry (MS) has emerged as a powerful technique to study protein glycosylation. MS on intact denatured or native proteins can reveal all-inclusive glycoproteoform profiles while top-down, middle-down and/or bottom-up MS can uncover the characteristics of individual glycosylation sites. Alternatively, analyzing enzymatically released N-glycans can reveal intricate details on glycan isomers and generate high-throughput data on larger cohorts. All these methods are increasingly applied for the study of both individual glycoproteins and complex glycoprotein mixtures such as those originating from blood plasma or cell lysates. This has increased our knowledge about the complexity of protein glycosylation, but also revealed its huge diversity, which depends not only on the protein but also on the cell-dependent glycosylation machinery that may change with physiological conditions. Currently, multiple glycoproteins are recombinantly produced, for therapeutic applications as well as in the food sector, in host cells of diverse origin, most commonly: E. coli bacteria, yeast cells, insect cells, mammalian CHO or human HEK293 cells. Although glycoproteins of interest might show similar yields when produced in different host cells, an important question remains whether the host cell will or can provide similar or alike glycoproteoform profiles. In this review, we focus on the application of MS-based technologies to study glycosylation profiles of endogenous human glycoproteins and their recombinantly produced counterparts in different host cells. We will discuss in which ways recombinant glycoproteins can differ from their endogenous variants, and the functional consequences.
