BioTech

27 pages, 2600 KB

Open AccessArticle

Valorization of Agave Leaf Juice for Optimized Kocuria sediminis AS04 Production and Its Delivery via Immobilized Films to Mitigate Saline Stress in Capsicum annuum var. glabriusculum

by Claudia Estefania Cabrera-Muro, Rosa María Camacho-Ruiz, Miguel Angel Lorenzo-Santiago, Jacobo Rodriguez-Campos and Silvia Maribel Contreras-Ramos

BioTech 2026, 15(3), 67; https://doi.org/10.3390/biotech15030067 - 15 Aug 2026

Abstract

Halotolerant plant growth-promoting bacteria offer a sustainable strategy to improve crop performance under saline conditions. This study evaluated Agave tequilana Weber var. Azul leaf juice was used as an alternative growth medium for Kocuria sediminis AS04, and the bacterium’s ability to alleviate salt [...] Read more.

Halotolerant plant growth-promoting bacteria offer a sustainable strategy to improve crop performance under saline conditions. This study evaluated Agave tequilana Weber var. Azul leaf juice was used as an alternative growth medium for Kocuria sediminis AS04, and the bacterium’s ability to alleviate salt stress in chiltepin (Capsicum annuum) was evaluated. The chiltepin seedlings were grown in a specialized chamber and exposed to higher salt levels (200, 400, and 600 mM NaCl) for 10 days. During this time, the protective effect of K. sediminis AS04, which was held in a polymer film, was examined. K. sediminis AS04 grew well in a medium containing 25% agave juice and urea, reaching a density of 1.3 × 10¹⁰ CFU mL⁻¹. Compared with conventional Tryptic Soy Broth, the medium formulated from agave leaf juice and urea could reduce the cost per kilogram of biomass by approximately 4-fold. Under severe salinity stress, plants inoculated with immobilized K. sediminis at 600 mM NaCl exhibited the highest shoot biomass (0.31 g⁻¹ plant), root length (50.7 mm), and proline accumulation (11.25 µmol g⁻¹ fresh weight), whereas uninoculated plants displayed reduced biomass (0.16 g plant⁻¹) and shorter roots (34.5 mm). At 600 mM NaCl, inoculation increased shoot biomass, root length, and plant survival by approximately 94%, 47%, and 200%, respectively, compared with uninoculated seedlings. This method values agro-industrial waste and enhances chiltepin performance under high-salinity conditions. It promotes sustainable production and helps create affordable inoculants for agricultural biotechnology. Full article

(This article belongs to the Section Industry, Agriculture and Food Biotechnology)

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24 pages, 11586 KB

Open AccessArticle

Risk-Aware Computational Prioritization and Validation Route Design for Medicine–Food Homology Plant Compounds in a Parkinson’s Disease Context

by Jinhao Zou, Siyi Wang, Jingjiao Yong, Liangyu Yan, Hong Hui and Ye Sun

BioTech 2026, 15(3), 66; https://doi.org/10.3390/biotech15030066 - 10 Aug 2026

Abstract

Network pharmacology studies of medicine–food homology plants have identified broad injury response pathways and hubs that cannot support compound-level or Parkinson’s disease (PD)-specific claims. We developed a traceable, non-weighted framework that separates regulatory provenance, PD-context evidence, structural support, and safety/developability liabilities. A ten-plant [...] Read more.

Network pharmacology studies of medicine–food homology plants have identified broad injury response pathways and hubs that cannot support compound-level or Parkinson’s disease (PD)-specific claims. We developed a traceable, non-weighted framework that separates regulatory provenance, PD-context evidence, structural support, and safety/developability liabilities. A ten-plant feasibility panel was locked before overlap with a 1631-gene PD union, yielding 382 plant-associated targets and 190 strict intersections. Leave-one-plant-out analysis retained 173–190 targets, whereas disease source and threshold stress tests showed curation dependence. Whole-blood classifiers showed modest five-fold discrimination (area under the curve, 0.606–0.682) and were excluded from candidate decisions. Redocking-validated AutoDock Vina and protein–ligand interaction fingerprints retained baicalein–MMP9, baicalein–AKT1, and baicalein–BCL2 as caution-tagged follow-up pairs. Quercetin–MMP9 was retained as a liability-tagged comparator, while KCNH2 relations were safety-only. Because no biological validation is presented, these pairs remain hypotheses for prospective MPP⁺-treated SH-SY5Y testing with orthogonal injury, dopaminergic phenotypes, target dependency, material confirmation and safety controls. Baicalein remains source-pending for the material chain. Full article

(This article belongs to the Section Computational Biology)

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18 pages, 2108 KB

Open AccessArticle

Effects of Orange Peel-Derived Carbon Sources on Nannochloropsis salina Mixotrophic Cultivation

by Carlo Esposito, Giancarlo Aldini, Yanan Yin, Stefano Gandolfi, Gianluca Ottolina and Francesco Secundo

BioTech 2026, 15(3), 65; https://doi.org/10.3390/biotech15030065 - 7 Aug 2026

Abstract

Microalgae are recognized as sustainable biofactories for metabolites relevant to nutraceutical, pharmaceutical, and marine drug applications. Nannochloropsis salina is notable for its production of bioactive lipids, including the pharmaceutically relevant omega-3 eicosapentaenoic acid (EPA). Here, we evaluated orange peel extract (OPE), a citrus [...] Read more.

Microalgae are recognized as sustainable biofactories for metabolites relevant to nutraceutical, pharmaceutical, and marine drug applications. Nannochloropsis salina is notable for its production of bioactive lipids, including the pharmaceutically relevant omega-3 eicosapentaenoic acid (EPA). Here, we evaluated orange peel extract (OPE), a citrus by-product, as a substrate for mixotrophic cultivation of N. salina within a bioeconomy framework. OPE supplementation triggered dose-dependent physiological responses. Among the tested OPE concentrations, 5% supplementation resulted in the highest total fatty acid content, increasing fatty acids from 24.4% (control) to 30.6% and EPA from 6.4% to 9.8%, whereas 10% OPE maintained biomass pigmentation. Higher OPE levels enhanced antioxidant potential, raising total antioxidant capacity from 4.6 (control) to 6.7 mg/g vitamin C equivalents (with 10% OPE), while the highest concentrations induced metabolic stress, reducing biomass and altering lipid composition. Fourier transform infrared spectroscopy analyses revealed biochemical adjustments consistent with metabolic reorganization, including increased intensities of the amide I and II bands associated with protein-rich structures. Overall, OPE emerges as a cost-effective supplement capable of modulating the biochemical quality of N. salina while valorizing agro-industrial residues. The increases in EPA and antioxidant capacity support the potential of OPE-supplemented cultures as a platform for the production of marine-derived bioactive compounds. Full article

(This article belongs to the Section Industry, Agriculture and Food Biotechnology)

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15 pages, 961 KB

Open AccessArticle

Night-Time Biomass and Compositional Dynamics in Chlorella vulgaris: Optimisation of Harvesting Time

by Sofia Pires, Susana Casal, Tânia G. Tavares, José C. M. Pires and Joana Oliveira

BioTech 2026, 15(3), 64; https://doi.org/10.3390/biotech15030064 - 6 Aug 2026

Abstract

Global population growth has emphasised the need to have sustainable and alternative sources of nutrients. In this context, microalgae have emerged as a potential solution due to their rich biochemical composition, including high-quality proteins, carbohydrates, lipids, and pigments. This study investigates the variation [...] Read more.

Global population growth has emphasised the need to have sustainable and alternative sources of nutrients. In this context, microalgae have emerged as a potential solution due to their rich biochemical composition, including high-quality proteins, carbohydrates, lipids, and pigments. This study investigates the variation in microalgal growth and biochemical composition over a light:dark cycle, with a focus on the night period. Batch experiments were performed with eight Chlorella vulgaris cultures over a 7-day period. On the seventh day, biomass samples were collected at four time points in four-hour intervals and stored for subsequent biochemical analyses. During the eight-hour dark period, biomass, carbohydrate, and total chlorophyll concentrations decreased by 9%, 12.5%, and 14.4%, respectively. After four hours of light exposure, these parameters increased significantly by 6%, 15.4%, and 12.7%, respectively. Total protein, carotenoids, and fatty acid contents remained relatively stable throughout the evaluated cycle, although variations were observed in the carotenoid profile. During the dark phase, zeaxanthin decreased by 38.0%, whereas violaxanthin increased by 27.2%, suggesting complementary pigment interconversion consistent with xanthophyll cycle activity. Overall, these results highlight the importance of optimising harvesting time to enhance the production of target compounds in a sustainable production of microalgal biomass. Full article

(This article belongs to the Section Environmental Biotechnology)

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21 pages, 1819 KB

Open AccessFeature PaperArticle

Differential Correlation Across Subpopulations of Single Cells in Subtypes of Acute Myeloid Leukemia

by Reginald L. McGee II, Jake Reed, Gregory K. Behbehani and Kevin R. Coombes

BioTech 2026, 15(3), 63; https://doi.org/10.3390/biotech15030063 - 5 Aug 2026

Abstract

Mass cytometers can record 40–50 parameters per single cell for millions of cells in a sample. Many methods have been developed to cluster phenotypically similar cells within cytometry data, but there are fewer methods to visualize activity and interactions of pairs of proteins [...] Read more.

Mass cytometers can record 40–50 parameters per single cell for millions of cells in a sample. Many methods have been developed to cluster phenotypically similar cells within cytometry data, but there are fewer methods to visualize activity and interactions of pairs of proteins across these populations. We have developed a workflow for analyzing correlations associated with predefined populations. By clustering blood samples from acute myeloid leukemia (AML) patients and normal controls using an established algorithm, we obtained a minimum spanning tree of clusters of single cells. Using surface marker expression, we identified clusters on the tree that belonged to phenotypes of interest. Next, we computed correlations between pairs of proteins in each cluster. We developed a novel, coherent, probability-based statistic to test differences between vectors of correlation coefficients. By comparing all combinations of the normal controls under the statistic, we created an empirical distribution that could provide a conservative threshold of differential correlation. Using this empirically derived distribution to define significance, we compared pooled samples from AML subtypes and normal controls to detect differential correlations. Given the structure present within this cytometry dataset, we found it natural to consider correlations in this manner versus aggregating all data and computing a single correlation. Our approach has the advantage that we can localize the statistical measure to determine contributions from particular phenotypic populations. Differentially correlated pairs of proteins can be further explored as possible testable hypotheses by considering a population’s distribution of correlation coefficients or biaxially plotting protein expressions within individual cells in a given population. Full article

(This article belongs to the Section Computational Biology)

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17 pages, 1168 KB

Open AccessArticle

Eisenia andrei and Tenebrio molitor Divergently Restructure the Bacteriome and Mycobiome of Sewage Sludge with Contrasting Biosafety Consequences

by Eduardo Mancilla, Marcos Pérez-Losada, Manuel Aira and Jorge Domínguez

BioTech 2026, 15(3), 62; https://doi.org/10.3390/biotech15030062 - 3 Aug 2026

Abstract

The use of invertebrates for sewage sludge bioconversion offers a sustainable strategy for waste valorization, yet species-specific effects on microbial communities and biosafety remain unclear. Here, we compared the impacts of the earthworm Eisenia andrei (Ea) and the mealworm Tenebrio molitor (Tm) on [...] Read more.

The use of invertebrates for sewage sludge bioconversion offers a sustainable strategy for waste valorization, yet species-specific effects on microbial communities and biosafety remain unclear. Here, we compared the impacts of the earthworm Eisenia andrei (Ea) and the mealworm Tenebrio molitor (Tm) on the bacteriomes and mycobiomes of sewage sludge (ss) using 16S rRNA and ITS amplicon sequencing. Gut passage in both Ea and Tm markedly altered bacterial and fungal composition relative to ss, but produced distinct community profiles with differential shifts across multiple taxa. Both invertebrates reduced bacterial richness by ~40%, while fungal responses diverged: Ea largely preserved mycobiome richness despite reduced evenness, whereas Tm caused a near-complete collapse (~83% Amplicon Sequence Variant loss). Beta diversity analyses revealed clear, non-overlapping separation among ss, Ea, and Tm for both microbial domains. Tm frass showed strong enrichment of clinically relevant bacterial pathogens, while Ea casts exhibited no such enrichment. For fungi, Ea reshaped pathogen composition, whereas Tm largely eliminated fungal pathogens through broad community collapse. Both treatments reduced predicted antibiotic resistance gene abundance, but functional profiles differed, with Ea showing greater functional stability. These findings demonstrate that microbiome restructuring during bioconversion is species-dependent, with contrasting ecological and biosafety implications for downstream environmental use. Full article

(This article belongs to the Special Issue Biotechnology for Wastewater Treatment: Current Situation and Future Prospects)

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22 pages, 3431 KB

Open AccessArticle

Detection of an ExPEC-like Escherichia coli ST536 Isolated from Depurated Retail Mussels: Genomic Insights into Food Safety and One Health Surveillance

by Ricardo J. Figueiredo, Guilherme Moreira, Ana Machado, Eliane Silva, Adriano A. Bordalo and João R. Mesquita

BioTech 2026, 15(3), 61; https://doi.org/10.3390/biotech15030061 - 31 Jul 2026

Abstract

Filter-feeding bivalves can accumulate fecal bacteria and antimicrobial resistance determinants, yet routine regulatory monitoring relies mainly on quantitative Escherichia coli criteria. This study provides a whole-genome characterization of an ExPEC-like E. coli ST536 (O21) isolate recovered from commercially depurated retail mussels originating from [...] Read more.

Filter-feeding bivalves can accumulate fecal bacteria and antimicrobial resistance determinants, yet routine regulatory monitoring relies mainly on quantitative Escherichia coli criteria. This study provides a whole-genome characterization of an ExPEC-like E. coli ST536 (O21) isolate recovered from commercially depurated retail mussels originating from a Class B harvesting area in Portugal. The isolate was phenotypically identified by MicroScan WalkAway Plus and subjected to antimicrobial susceptibility testing and Oxford Nanopore long-read sequencing, followed by genome assembly, annotation, antimicrobial resistance and virulence screening, plasmid and prophage detection, and phylogenomic analysis. Phenotypically, only resistance to tobramycin was detected using the MicroScan WalkAway system. Genomic analysis revealed a complex mobile element-rich architecture, including multiple resistance-associated determinants, 116 virulence-associated loci, prophage regions, and a mobilizable IncY plasmid carrying CTX-like class A β-lactamase-associated hits detected by ABRicate. Despite its ST536 assignment and ExPEC-associated traits, phylogenomic analysis placed the isolate within a phylogroup A/commensal-like background rather than among classical B2 ExPEC lineages, suggesting acquisition of pathoadaptive modules by an atypical environmental lineage. These findings highlight the potential utility of WGS as a complementary tool and suggest that depurated bivalves may serve as sentinels, warranting further investigation. Full article

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22 pages, 4186 KB

Open AccessFeature PaperArticle

Germination-Induced Variations in Proximate Composition, Phytochemical Profile and Antioxidant Activities of White Sesame Seeds: A Green Approach for Nutritional Modification

by Oneeza Anwar, Iahtisham-Ul-Haq, Ramiz Arif, Waqas Ahmed and Hafiz Ansar Rasul Suleria

BioTech 2026, 15(3), 60; https://doi.org/10.3390/biotech15030060 - 29 Jul 2026

Abstract

Germination is a cost-effective bioprocessing technique for enhancing seed properties such as sesame seeds, an underutilized seed crop. This research evaluates modifications in physicochemical and antioxidant activity of white sesame seeds (Sesamum indicum L.) influenced by germination. In this context, non-germinated seeds [...] Read more.

Germination is a cost-effective bioprocessing technique for enhancing seed properties such as sesame seeds, an underutilized seed crop. This research evaluates modifications in physicochemical and antioxidant activity of white sesame seeds (Sesamum indicum L.) influenced by germination. In this context, non-germinated seeds and germinated seeds at 24, 48, 72, 96, and 120 h were evaluated separately for their phytochemical profile, including TFC (aluminum chloride colorimetric method), TPC (Folin–Ciocalteu method) and antioxidant properties (DPPH and ABTS radical scavenging assays). Significant variations in the proximate composition, phytochemical densities, and antioxidant activities of white sesame seeds were observed as influenced by germination. The statistical analysis showed an increase in flavonoid content when germination was extended to 48 h, but a decline when extended beyond 48 h, whereas higher polyphenol quantities were observed in sesame seeds germinated for 96 and 120 h. The DPPH activity of germinated (24 h) sesame seed extracts was highest. In contrast, ABTS activity was higher in non-germinated seeds than in seeds germinated for 96 h, and it began to decline at 120 h. In general, a negative correlation between antioxidant activity and extended germination time (120 h) has been observed. For the incorporation of sesame seeds into functional food products, 48 h of germination appeared to have potential suitability for a favorable antioxidant and phytochemical profile. Full article

(This article belongs to the Collection Natural Antioxidants in Food and Nutraceuticals: Methodological Innovations, Bioavailability and Health Mechanisms)

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21 pages, 9567 KB

Open AccessArticle

Establishment of a High-Efficiency In Vitro Regeneration and Agrobacterium-Mediated Genetic Transformation System for Ajania achilleoides (Turcz.) Poljakov

by Xiaoyue Zheng, Hao Li, Shu Wang, Huiyan Cheng and Huien Zhao

BioTech 2026, 15(3), 59; https://doi.org/10.3390/biotech15030059 - 28 Jul 2026

Abstract

Ajania achilleoides, a wild species with strong environmental adaptability, is a valuable germplasm resource for genetic improvement and functional gene studies. However, efficient in vitro regeneration and genetic transformation systems for this species remain undeveloped, limiting functional gene characterization and genetic improvement [...] Read more.

Ajania achilleoides, a wild species with strong environmental adaptability, is a valuable germplasm resource for genetic improvement and functional gene studies. However, efficient in vitro regeneration and genetic transformation systems for this species remain undeveloped, limiting functional gene characterization and genetic improvement of this valuable germplasm resource. In this study, leaf explants were used to establish an efficient regeneration system by evaluating different combinations of plant growth regulators and to develop an Agrobacterium tumefaciens-mediated genetic transformation system for A. achilleoides. MS medium supplemented with 2.0 mg/L 6-benzyladenine (6-BA) and 1.0 mg/L α-naphthaleneacetic acid (NAA) achieved the highest adventitious shoot regeneration rate (99%), while 1/2 MS medium containing 0.5 mg/L indole-3-butyric acid (IBA) was optimal for rooting, resulting in a 100% rooting rate. Critical hygromycin concentrations for shoot regeneration and rooting selection were determined to be 25 mg/L and 10 mg/L, respectively. For transformation, the best results were obtained with a 3-day preculture, bacterial suspension at OD₆₀₀ = 0.2, 30 min infection, and 2-day co-cultivation. PCR and sequencing confirmed the successful integration of the target genes into the genome, yielding three positive transgenic lines with a molecular confirmation rate of 30%. This study establishes a stable and efficient regeneration and transformation system for A. achilleoides, providing a platform for functional gene analysis and molecular breeding of wild chrysanthemum species. Full article

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37 pages, 21317 KB

Open AccessReview

Omics Approaches in Hantavirus Research: Current Advances, Challenges, and Future Perspectives

by Soroosh Najafi, Maryam Jojani, Kianoosh Najafi and Giovanni N. Roviello

BioTech 2026, 15(3), 58; https://doi.org/10.3390/biotech15030058 - 23 Jul 2026

Abstract

Hantaviruses are zoonotic RNA viruses responsible for two severe human diseases: hemorrhagic fever with renal syndrome (HFRS) and hantavirus cardiopulmonary syndrome (HCPS). Worldwide case fatality rates vary from 1% to 40%. Although single-layer genomics, transcriptomics, proteomics, and metabolomics studies have advanced our understanding [...] Read more.

Hantaviruses are zoonotic RNA viruses responsible for two severe human diseases: hemorrhagic fever with renal syndrome (HFRS) and hantavirus cardiopulmonary syndrome (HCPS). Worldwide case fatality rates vary from 1% to 40%. Although single-layer genomics, transcriptomics, proteomics, and metabolomics studies have advanced our understanding of hantavirus biology, true multi-omics integration remains scarce, leaving systems-level mechanisms of disease severity and host–pathogen interactions unresolved. High-throughput omics technologies have greatly advanced the study of hantavirus–host interactions. This review summarizes genomic, transcriptomic, proteomic, metabolomic, and AI-enabled approaches in hantavirus research. Genomic studies have clarified viral diversity, evolution, and reassortment, while transcriptomics has identified regulatory networks governing endothelial and innate immune responses. Proteomics has revealed host proteins involved in immune regulation, endothelial dysfunction, and potential therapeutic targeting, whereas metabolomics indicates substantial metabolic reprogramming during infection, although dedicated studies remain limited. AI-based approaches are increasingly applied to outbreak prediction, surveillance, and risk modeling. Drawing on successful multi-omics frameworks developed for other viral infections, we discuss opportunities to improve biomarker discovery, surveillance, therapeutic development, and future precision medicine strategies for hantavirus infections. Full article

(This article belongs to the Special Issue Integrative Omics Approaches for Precision Biotech: Tools, Applications and Future Perspectives)

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14 pages, 1729 KB

Open AccessArticle

Psychrophilic Quorum Sensing Genes Enable Unimodal, Adjustable Protein Expression Across the Entire Escherichia coli Population

by Ekaterina Scheglova, Sabina Nebieva, Kamilla Mekhantseva, Siarhei Bukhalovich, Anna Kudryavtseva, Nikolay Bondarev, Nikolay Ilyinsky, Sergey Bazhenov and Ilya Manukhov

BioTech 2026, 15(3), 57; https://doi.org/10.3390/biotech15030057 - 21 Jul 2026

Abstract

A previous study demonstrated that quorum sensing (QS) genes from the psychrophilic bacterium Aliivibrio logei can be used in Escherichia coli to obtain bacterial cultures with controlled amounts of a target protein. However, the QS system may be induced non-uniformly across the cell [...] Read more.

A previous study demonstrated that quorum sensing (QS) genes from the psychrophilic bacterium Aliivibrio logei can be used in Escherichia coli to obtain bacterial cultures with controlled amounts of a target protein. However, the QS system may be induced non-uniformly across the cell population. In this study, we used an expression vector containing luxR/luxI regulatory genes from A. logei carrying sfGFP as a reporter gene. Reporter expression was regulated by the autoinducer 3OxoC6-HSL, activated at 22 °C, and terminated at 37 °C. Flow cytometry was used to assess the GFP fluorescence distribution at the single-cell level. The system provided dose-dependent unimodal expression lacking formation of distinct ON/OFF subpopulations, while the robust coefficient of variation decreased with increasing autoinducer concentration. The autoinducer synthase LuxI enabled autoinduction, but under the tested conditions, no substantial effect on expression homogeneity was detected. Raising the temperature to 37 °C effectively halted expression, allowing intermediate target protein values to be fixed at the single-cell level. Overall, the developed system represents a promising tool for biotechnological applications requiring precise and uniform control of expression. Full article

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30 pages, 2883 KB

Open AccessReview

Advances in Betalain Biosynthesis and Metabolic Engineering for Sustainable Natural Pigment Production

by Saravanan Monisha, Marimuthu Kanchana, Aiyar Balasubramanian and Rajendran K. Selvakesavan

BioTech 2026, 15(3), 56; https://doi.org/10.3390/biotech15030056 - 19 Jul 2026

Abstract

Betalains are water-soluble pigments containing nitrogen, and they exist naturally in the plants of the order Caryophyllales. They have gained increasing attention in recent years because of their intense colours, antioxidant activity, and safety, thus making them suitable replacements for artificial dyes. The [...] Read more.

Betalains are water-soluble pigments containing nitrogen, and they exist naturally in the plants of the order Caryophyllales. They have gained increasing attention in recent years because of their intense colours, antioxidant activity, and safety, thus making them suitable replacements for artificial dyes. The increasing interest in natural pigments has led to intensified research on betalain biosynthesis and optimization of pigment production. Nonetheless, their application in industry faces limitations, such as their low natural occurrence, sensitivity to environmental conditions, and instability during manufacturing and storage. Unlike previous reviews that primarily focused on betalain chemistry, biosynthesis pathways, or biological activity, the present review highlights recent developments in the engineering of the biosynthesis pathways, synthetic biology, elicitation approaches, omics-based pathway identification, and nanobiotechnology for betalain pigments. Special attention is paid to the comparison of plant, plant cell, yeast, and bacterial production systems, as well as recent advancements towards industrial production of betalain pigments and bottlenecks in the commercialization of sustainable betalain bio-factories. Full article

(This article belongs to the Section Industry, Agriculture and Food Biotechnology)

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22 pages, 3869 KB

Open AccessArticle

AAV Vector Toolkit for the Delivery and Expression of the Artificial microRNA in the Murine Heart

by Ivan I. Galkin, Viktoriia V. Skopenkova, Maria Y. Shubina, Anna V. Polikarpova, Svetlana G. Vassilieva, Irina M. Savchenko, Olga S. Lebedeva, Daria V. Goliusova, Margarita Y. Sharikova, Vladimir V. Gureev, Tatiana N. Malorodova, Alexey V. Deikin, Tatiana V. Egorova and Maryana V. Bardina

BioTech 2026, 15(3), 55; https://doi.org/10.3390/biotech15030055 - 17 Jul 2026

Abstract

Background: Adeno-associated virus (AAV) vector-mediated gene transfer is an emerging treatment strategy for severe cardiac disorders with genetic etiology. We refined the AAV toolkit to achieve efficient and selective expression of the therapeutic transgene in mouse hearts. Methods: Using vectors with a reporter [...] Read more.

Background: Adeno-associated virus (AAV) vector-mediated gene transfer is an emerging treatment strategy for severe cardiac disorders with genetic etiology. We refined the AAV toolkit to achieve efficient and selective expression of the therapeutic transgene in mouse hearts. Methods: Using vectors with a reporter transgene, we evaluated AAV administration routes, AAV serotype tropism to the myocardium, and cardiospecific promoters. Results: We showed that systemic AAV administration provides potent delivery and uniform transduction of cardiac tissue, outperforming localized injection techniques. The MyoAAV 2A capsid variant enabled an improved heart-to-liver transduction ratio compared to the parental AAV9 serotype. Screening a panel of cardiac and pan-muscular promoters in vitro and in vivo verified the superiority of the cardiac troponin T (cTnT) promoter for robust heart-specific transgene expression. Finally, we demonstrated that the cumulative properties of systemic AAV delivery, the MyoAAV 2A serotype, and the cTnT promoter allowed for efficient cardiac synthesis of the therapeutic transgene—an artificial miRNA designed for the gene suppression strategy of FLNC-related cardiomyopathy. Conclusions: Our findings establish an effective AAV approach for transgene transfer into the mouse heart and promote the development of gene therapy for cardiac disorders. Full article

(This article belongs to the Topic Advances in Gene Therapy of Human Diseases)

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21 pages, 5922 KB

Open AccessReview

Extracellular Matrix-Based and Extracellular Matrix-Bioinspired Scaffolds for Extracellular Vesicle Delivery in Dental Pulp Regeneration: A Narrative Review

by Nevena Cvetic, Anđelka Zivaljevic, Kristina Krstic, Suzana Zivanovic, Milos Papic, Natalija Arsenijevic, Miona Glisic, Tamara Milunovic, Renata Petrovic and Milica Popovic

BioTech 2026, 15(3), 54; https://doi.org/10.3390/biotech15030054 - 14 Jul 2026

Abstract

Vital pulp therapy aims to preserve pulp vitality by stimulating reparative processes. However, conventional approaches often result in incomplete tissue regeneration. Extracellular vesicles (EVs) have emerged as promising cell-free therapeutic agents because of their ability to regulate angiogenesis, odontogenesis, and immune responses through [...] Read more.

Vital pulp therapy aims to preserve pulp vitality by stimulating reparative processes. However, conventional approaches often result in incomplete tissue regeneration. Extracellular vesicles (EVs) have emerged as promising cell-free therapeutic agents because of their ability to regulate angiogenesis, odontogenesis, and immune responses through the transfer of bioactive molecules. Despite their significant regenerative potential, the clinical application of EVs remains limited by rapid clearance, insufficient local retention, and uncontrolled release following administration. To address these challenges, various extracellular matrix (ECM)-based and ECM-bioinspired scaffolds have been developed as delivery platforms. These scaffolds can provide structural support and enable controlled, localized release of EVs. This narrative review critically evaluates the current evidence regarding scaffold systems as EV delivery platforms for dental pulp regeneration, comparing their biological performance, methodological quality, and translational potential. Across the available studies, scaffold-assisted EV delivery consistently enhanced angiogenesis, odontogenic differentiation, mineralization, and immunomodulation; however, the evidence remains preclinical and is characterized by substantial heterogeneity regarding EV source, isolation and characterization methods, scaffold composition, experimental models, and outcome assessment. Current findings support the feasibility of scaffold-assisted EV delivery for regenerative endodontics, but important challenges remain, including standardization of EV production and characterization, scalable manufacturing, regulatory approval, and demonstration of long-term safety and functional pulp–dentin complex regeneration. Further well-designed translational and clinical studies will be essential before routine clinical implementation can be considered. Full article

(This article belongs to the Section Medical Biotechnology)

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14 pages, 1292 KB

Open AccessArticle

RNA-seq Co-Expression Analysis Reveals a Midgut-Associated Digestive Gene Module in Helicoverpa armigera

by Bairon J. Matabanchoy Pejendino, Vicente E. Mallama Cadena, María C. Díaz Rodríguez, Claudia Salazar Gonzalez and Pedro A. Velasquez-Vasconez

BioTech 2026, 15(3), 53; https://doi.org/10.3390/biotech15030053 - 13 Jul 2026

Abstract

Helicoverpa armigera is one of the most destructive polyphagous pests, yet the transcriptional organization underlying its digestive capacity remains poorly resolved. Here, we compiled 579 publicly available RNA-seq libraries representing 54 independent experiments and quantified transcript abundance across tissues and developmental stages. This [...] Read more.

Helicoverpa armigera is one of the most destructive polyphagous pests, yet the transcriptional organization underlying its digestive capacity remains poorly resolved. Here, we compiled 579 publicly available RNA-seq libraries representing 54 independent experiments and quantified transcript abundance across tissues and developmental stages. This complete dataset was used to support broader tissue-level expression profiling. After metadata harmonization and quality filtering, a subset of 130 biologically comparable libraries from five tissue/developmental categories was retained for weighted gene co-expression network analysis. WGCNA identified four biologically informative modules, among which the turquoise module was positively associated with fourth- and fifth-instar larval midgut samples. Independent expression profiling revealed strong midgut-biased expression of several trypsin- and chymotrypsin-like serine proteases, although only a subset of these genes was assigned to the turquoise module. Descriptive functional annotation of this module identified 202 co-expressed loci, including digestive enzymes, nutrient transporters, detoxification-related proteins, epithelial components and putative transcriptional or signaling-associated genes. Phylogenetic analyses and manual inspection of genomic locations further showed that several digestive protease genes occur in local clusters and have closely related counterparts in H. zea, suggesting partial conservation of local genomic organization. Collectively, these results describe a midgut-associated co-expression module containing genes associated with digestive, absorptive and protective functions and provide candidate genes for future functional studies. Full article

(This article belongs to the Special Issue The Emerging Role of Bioinformatics in Biotechnology)

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21 pages, 4322 KB

Open AccessArticle

Global Patent Landscape and Technological Trends in Biosafety Level 3 (BSL-3) Laboratories Technologies

by Milca de J. Silva, Roni D. Vinhas, Helena S. da Hora, Saada L. C. Fernandez, Hayna Malta-Santos, Hugo Saba, Camila D. F. Ribeiro, Marilda de S. Gonçalves and Bruna A. S. Machado

BioTech 2026, 15(3), 52; https://doi.org/10.3390/biotech15030052 - 10 Jul 2026

Abstract

Biosafety Level 3 (BSL-3) laboratories are essential for handling high-risk pathogens and strengthening global health security. This study presents a patent landscape analysis of BSL-3-related technologies using the Derwent World Patents Index (DWPI) to identify technological trends, geographic distribution, patent classifications, and temporal [...] Read more.

Biosafety Level 3 (BSL-3) laboratories are essential for handling high-risk pathogens and strengthening global health security. This study presents a patent landscape analysis of BSL-3-related technologies using the Derwent World Patents Index (DWPI) to identify technological trends, geographic distribution, patent classifications, and temporal evolution. Patent documents associated with laboratory infrastructure, ventilation systems, containment devices, and biosafety procedures were screened and analyzed. A total of 58 patent documents filed between 2009 and 2024 met the inclusion criteria. The results showed that China and the United States are the leading contributors to BSL-3 patent development, reflecting continued investments in biosafety and biosecurity infrastructure. The most frequent International Patent Classification (IPC) categories were C12M (microbiological devices), E04H (specialized construction infrastructure), and F24F (ventilation and air control systems), highlighting the multidisciplinary nature of innovations in laboratory containment and safety. The temporal trends revealed increases in patent activity following major public health emergencies, including SARS, Ebola, and particularly the COVID-19 pandemic. Furthermore, a significant increase in patent expirations is expected by 2029, creating opportunities for technology transfer, open innovation, and broader access to critical biosafety technologies. These findings emphasize the strategic importance of continued investment in BSL-3 technologies, especially in developing countries with growing biosafety demands. Full article

(This article belongs to the Section Biotechnology Regulation)

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16 pages, 3701 KB

Open AccessArticle

Stabilizing Nanoemulsions with Blended Biosurfactants: Role of Sophorolipids and Lecithin in Emulsion Performance

by Yew Seng Leow, Dayang Radiah Awang Biak, Nur Syakina Jamali, Huey Fang Teh and Norhafizah Abdullah

BioTech 2026, 15(3), 51; https://doi.org/10.3390/biotech15030051 - 2 Jul 2026

Abstract

Sophorolipids (SLs) produced from Starmerella bombicola using four different secondary substrates such as refined, bleached, and deodorized palm olein (RBD PO), RBD palm kernel olein (RBD PKO), RBD coconut olein (RBD CO) and fatty acid methyl ester (FAME) waste are reported. Their interfacial [...] Read more.

Sophorolipids (SLs) produced from Starmerella bombicola using four different secondary substrates such as refined, bleached, and deodorized palm olein (RBD PO), RBD palm kernel olein (RBD PKO), RBD coconut olein (RBD CO) and fatty acid methyl ester (FAME) waste are reported. Their interfacial characteristics at medium-chain triglyceride (MCT) oil-water interface and ability to form nano/submicron emulsions were studied. The effects of SLs from different sources, SL concentrations and blend ratios of SLs and soybean lecithin on characteristics of emulsions produced by ultrasonication were examined. Initially, emulsion formed using SLs coded (from F2 to F5) showed large droplets (d₃₂ > 1000 nm) and poor stability. They were then blended with soybean lecithin at a ratio of 3:1 to produce emulsions coded F6 to F9 with smaller droplets (d₃₂ < 400 nm) and great stability over a range of temperatures (from 40 °C to 90 °C) and pH values (from 3 to 9). However, highly acidic (pH 2) and low ionic strength (1 mM NaCl) processing caused the separation of the emulsions. These emulsions also displayed potential antimicrobial activities towards Bacillus cereus and Pseudomonas aeruginosa, as well as cytotoxic effects against the human epithelial colorectal adenocarcinoma cell line (Caco-2). These results illustrated that stable emulsions required a mixture of SLs and soybean lecithin. Full article

(This article belongs to the Section Industry, Agriculture and Food Biotechnology)

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23 pages, 2921 KB

Open AccessReview

Next-Generation Metabolic Engineering of Capsaicinoids Biosynthesis in Chilli Pepper: Bridging Genomic Insights to Biotechnological Applications

by Thumadath Palayullaparambil Ajeesh Krishna, Deepa Harikrishnan, Mathew Veena, Theivanayagam Maharajan, M. James, Minisha Udhayakumar, Parimala Gnana Soundari Arockiam Jeyasundar, Sherrie Jesulyn David, Ramar Dineshkumar, Reshma Rajan and Periyasamy Rathinapriya

BioTech 2026, 15(3), 50; https://doi.org/10.3390/biotech15030050 - 1 Jul 2026

Abstract

Chilli peppers (Capsicum species) have been widely used around the world because of their economic value and distinctive sensory characteristics. They contain abundant functional metabolites, especially a group of vanillylamide compounds belonging to the family of capsaicinoids, which have been exploited for [...] Read more.

Chilli peppers (Capsicum species) have been widely used around the world because of their economic value and distinctive sensory characteristics. They contain abundant functional metabolites, especially a group of vanillylamide compounds belonging to the family of capsaicinoids, which have been exploited for medicinal, nutritional, agricultural, and cosmetic uses. The demand for capsaicinoid molecules is increasing day by day due to their high economic value and wide range of applications. Therefore, increasing bioactive metabolites, especially capsaicinoids in chilli peppers, is a major priority in the current scenario. Multi-omics approaches such as genomics, transcriptomics, proteomics, and metabolomics have substantially contributed to understanding the complex regulatory networks governing capsaicinoid biosynthesis. Key structural genes, transcription factors, and signaling pathways involved in the phenylpropanoid and branched-chain fatty acid pathways have been identified, providing valuable targets for metabolic engineering in chilli pepper. Despite these advances, the integration of genetic modification approaches for the targeted enhancement of capsaicinoid production remains limited in chilli pepper. Recent developments in biotechnology, particularly CRISPR/Cas-mediated genome-editing, enable the precise genetic modification of metabolic pathways and regulatory networks in plants. Therefore, it can contribute to the precise modification of key genes involved in the capsaicinoid biosynthesis pathway, offering potential strategies to enhance the capsaicinoid content in chilli pepper. However, CRISPR/Cas-mediated genome editing in chilli pepper is still in its early stages. There are currently no reports available on the successful enhancement of capsaicinoid content in chilli peppers through CRISPR/Cas-mediated genome editing. To date, no comprehensive review has evaluated the CRISPR-Cas-mediated genome-editing approaches for capsaicinoid metabolic engineering in chilli pepper. This review critically evaluates the recent advances in CRISPR/Cas–mediated metabolic engineering in chilli peppers, with particular emphasis on regulatory genes involved in capsaicinoid biosynthesis. Furthermore, multi-omics approaches are expected to complement these strategies by enabling the identification of key regulatory genes, the optimization of genome-editing targets, and the prediction of metabolic outcomes for enhanced capsaicinoid production. Overall, this review provides insights into improving capsaicinoid accumulation in chilli peppers through advanced genome-editing technologies. Full article

(This article belongs to the Section Industry, Agriculture and Food Biotechnology)

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32 pages, 1662 KB

Open AccessReview

Current Characterization Techniques Applied to Microalgae–Fungal Pellets: Unraveling the Mechanisms of Adhesion and Stability Focused on Nutrient Recovery/Recycling and Bioprocess Diversification

by João Victor Oliveira Nascimento da Silva, Carlos Eduardo de Farias Silva, Tomás Agustín Rearte, Eleni Kougia, Giorgos Markou and Albanise Enide da Silva

BioTech 2026, 15(3), 49; https://doi.org/10.3390/biotech15030049 - 29 Jun 2026

Abstract

Microalgae–fungal pellets have been studied as a versatile and robust biotechnological platform, offering significant advantages for microalgal biomass harvesting, wastewater treatment, biofuels production and/or obtaining of value-added products. This review presents an integrated analysis of the mechanisms governing the formation, stability, and functionality [...] Read more.

Microalgae–fungal pellets have been studied as a versatile and robust biotechnological platform, offering significant advantages for microalgal biomass harvesting, wastewater treatment, biofuels production and/or obtaining of value-added products. This review presents an integrated analysis of the mechanisms governing the formation, stability, and functionality of these systems, combining physicochemical, biological, and mathematical modelling approaches and aims to describe the current state of the art and main research needs. The aggregation process is strongly influenced by the complementarity of the surface properties of microalgae and filamentous fungi, including electrostatic interactions, production of extracellular polymeric substances (EPSs), and modifications in surface roughness. Recent advances in multiscale characterization techniques, such as confocal microscopy, micro-computed tomography, atomic force microscopy, and X-ray photoelectron spectroscopy, have allowed a more precise elucidation of the internal architecture and surface chemistry of the pellets. In parallel, biological characterization through enzymatic assays, oxidative stress biomarkers, and photosynthetic activity analyses has provided relevant information on the metabolic responses and functional resilience of the consortium. Additionally, the incorporation of mathematical flocculation models can contribute to the prediction of pellet growth, density, and stability, supporting process optimization and application. The understanding of these interaction phenomena is important for the design of high-yield and efficient systems, including their development and validation, to expand the use of microalgae–fungal pellets in bioprocesses, as evidenced by this review. Full article

(This article belongs to the Section Environmental Biotechnology)

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