BioTech

13 pages, 1451 KB

Open AccessArticle

Exposure to Sulfur Hexafluoride Influences Viability in Cell Transplant Suspensions

by Laura Martínez-Alarcón, Sergio Liarte, Juana M. Abellaneda, Juan J. Quereda, Livia Mendonça, Antonio Muñoz, Pablo Ramírez and Guillermo Ramis

BioTech 2025, 14(4), 86; https://doi.org/10.3390/biotech14040086 (registering DOI) - 31 Oct 2025

Abstract

Cell transplantation is often performed with ultrasonographic guidance for accurate delivery through injection. In such procedures, using ultrasonographic contrast greatly improves target delivery. However, accumulating evidence suggests that exposure to such contrast agents may have negative effects on transplanted cells. No study so [...] Read more.

Cell transplantation is often performed with ultrasonographic guidance for accurate delivery through injection. In such procedures, using ultrasonographic contrast greatly improves target delivery. However, accumulating evidence suggests that exposure to such contrast agents may have negative effects on transplanted cells. No study so far has researched this issue. Stabilized sulfur hexafluoride (SF6) microbubbles are a widely used sonographic contrast agent. Skin hCD55 porcine transgenic fibroblasts and mesenchymal stem cells from human bone marrow (hMSCs) were exposed in vitro to SF6 in concentrations ranging from 1.54 µM to 308 µM. The effects on viability and cell growth were registered using an impedance-based label-free Real-Time Cell Analyzer (RTCA). Data was recorded every 15 min for 50 h of total study time. Both cell lines behave distinctly when exposed to SF6. Porcine fibroblast growth showed relevant alterations only when exposed to higher concentrations. In contrast, hMSCs showed progressive growth decrease in relation to SF6 concentration. Taken together, while SF6-based contrast agents pose no threat to patient safety, our results indicate that exposure of suspended stem cells to the contrast agent could affect the effective dose administered in cell therapy procedures. This prompts specific cell lineage testing, adjusting methods and properly compensating for cell loss, with a potential impact on procedural cost and success rates. Full article

(This article belongs to the Section Medical Biotechnology)

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12 pages, 2048 KB

Open AccessArticle

Recombinant Clostridium acetobutylicum Endoxylanase for Xylooligosaccharide Production from Pretreated Lignocellulosic Biomass

by Afifa Husna, Agustin Krisna Wardani, Chun-Yi Hu and Yo-Chia Chen

BioTech 2025, 14(4), 85; https://doi.org/10.3390/biotech14040085 - 30 Oct 2025

Abstract

Xylooligosaccharides (XOS) are functional oligosaccharides with recognized prebiotic properties and growing industrial relevance, typically obtained through enzymatic depolymerization of xylan-rich lignocellulosic substrates. In this study, a recombinant endo-β-1,4-xylanase (XynA) from Clostridium acetobutylicum was employed for XOS production. The xynA gene was cloned into [...] Read more.

Xylooligosaccharides (XOS) are functional oligosaccharides with recognized prebiotic properties and growing industrial relevance, typically obtained through enzymatic depolymerization of xylan-rich lignocellulosic substrates. In this study, a recombinant endo-β-1,4-xylanase (XynA) from Clostridium acetobutylicum was employed for XOS production. The xynA gene was cloned into the expression vector pET-21a(+) and heterologously expressed in Escherichia coli BL21(DE3) under induction with isopropyl β-D-1-thiogalactopyranoside (IPTG). The recombinant protein, with an estimated molecular mass of 37.5 kDa, was verified by SDS-PAGE and Western blot analysis. Functional characterization via thin-layer chromatography revealed that XynA efficiently hydrolyzed beechwood xylan and rye arabinoxylan, predominantly yielding xylobiose. Additionally, the enzyme catalyzed the conversion of xylotriose into xylobiose and trace amounts of xylose. Notably, XynA demonstrated hydrolytic activity against autohydrolysed and alkali-pretreated coconut husk biomass, facilitating the release of XOS. These results underscore the potential of C. acetobutylicum XynA as a biocatalyst for the valorization of lignocellulosic residues into high-value oligosaccharides. Full article

(This article belongs to the Special Issue BioTech: 5th Anniversary)

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12 pages, 827 KB

Open AccessCommunication

Enhanced Succinate Production in Actinobacillus succinogenes via Neutral Red Bypass Reduction in a Novel Bioelectrochemical System

by Julian Tix, Fernando Pedraza, Roland Ulber and Nils Tippkötter

BioTech 2025, 14(4), 84; https://doi.org/10.3390/biotech14040084 - 29 Oct 2025

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Abstract

Carbon capture and power-to-X are becoming increasingly relevant in the context of decarbonization and supply security. Actinobacillus succinogenes is capable of transforming CO₂ into succinate, whereby product formation is significantly limited by the availability of NADH. The aim of this work was [...] Read more.

Carbon capture and power-to-X are becoming increasingly relevant in the context of decarbonization and supply security. Actinobacillus succinogenes is capable of transforming CO₂ into succinate, whereby product formation is significantly limited by the availability of NADH. The aim of this work was to further develop a bioelectrochemical system (BES) in order to provide additional reduction equivalents and thus increase yield and titer. To this end, a new BES configuration was established. A conventional stirred tank reactor (STR) is coupled via a bypass to an H-cell, in which the redox mediator neutral red (NR) is electrochemically reduced and then returned back to the bioreactor. The indirect electron transfer decouples the electrochemical reduction from the biology and results in increased intracellular availability of NADH. The present approach resulted in an increase in yield from 0.64 g·g⁻¹ to 0.76 g·g⁻¹, corresponding to an increase of 18%. At the same time, a titer of 16.48 ± 0.19 g·L⁻¹ was achieved in the BES, compared to 12.05 ± 0.18 g·L⁻¹ in the control. The results show that the mediator-assisted, partially decoupled BES architecture significantly improves CO₂-based succinate production and opens up a scalable path to the use of renewable electricity as a reduction source in power-to-X processes. Full article

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15 pages, 3966 KB

Open AccessArticle

Preparation of Suaeda Tea Through Semi-Solid Fermentation Utilizing Kluyveromyces marxianus, Komagataeibacter europaeus, and Acetobacter schutzenbachii: Physicochemical Characteristics, Process Optimization, and Antioxidant Activity

by Aoqi Dong, Xiaoying Dong, Xinying Dai, Yanru Gao, Yuewen Ning, Xiya Fan and Haiyan Liu

BioTech 2025, 14(4), 83; https://doi.org/10.3390/biotech14040083 - 28 Oct 2025

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Abstract

Suaeda salsa, an annual herb belonging to the genus Suaeda within the Chenopodiaceae family, is highly salt-tolerant and can thrive in large quantities on saline and alkaline soils. This study presents a novel fermentation technique to produce Suaeda tea, utilizing a synergistic [...] Read more.

Suaeda salsa, an annual herb belonging to the genus Suaeda within the Chenopodiaceae family, is highly salt-tolerant and can thrive in large quantities on saline and alkaline soils. This study presents a novel fermentation technique to produce Suaeda tea, utilizing a synergistic blend of microbial agents: Kluyveromyces marxianus, Komagataeibacter europaeus, and Acetobacter schutzenbachii. The resulting tea demonstrates a potent antioxidant capacity, with a hydroxyl radical scavenging rate of 64.2% and an exceptional 1,1-diphenyl-2-picrylhydrazyl radical scavenging capacity of 83.3%, along with increased ferric ion reduction/antioxidant power (FRAP) reducing power (1.82), indicating its superior antioxidant profile. Through the comparison of different microbial strain combinations under varying process parameters such as fermentation temperature and duration, the experiment revealed that fermentation at 37 °C for 24 h results in the highest concentrations of tea polyphenols (TPs) (≥10.87 mg/mL) and free amino acids (26.32 mg/100 mL). The quality of the fermented Suaeda tea meets the stringent GB/T 21733-2008 standards for tea beverages, exhibiting excellent physicochemical indices and sensory attributes. The antioxidant efficacy of the fermented Suaeda tea persists significantly throughout a 180-day duration. The optimization of the fermentation process for Suaeda tea not only provides a theoretical framework for large-scale production but also establishes a foundation for Suaeda salsa in the tea beverage sector. This innovation enriches the market with a diverse range of health-promoting teas, catering to the growing consumer demand for nutritious and beneficial beverages. Full article

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23 pages, 3638 KB

Open AccessArticle

Extracts of Argemone mexicana L. Contain Antifungal Compounds for the In Vitro Control of Monilinia fructicola, Colletotrichum gloeosporioides, Fusarium oxysporum, and Sclerotinia sclerotiorum: Preliminary Evidence for Field Application

by Iridiam Hernández-Soto, Antonio Juárez-Maldonado, Alfredo Madariaga-Navarrete, Ashutosh Sharma, Antonio de Jesus Cenobio-Galindo, Jose Manuel Pinedo-Espinoza, Aracely Hernández-Pérez and Alma Delia Hernández-Fuentes

BioTech 2025, 14(4), 82; https://doi.org/10.3390/biotech14040082 - 26 Oct 2025

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Abstract

Argemone mexicana L. is considered a weed; however, it contains secondary metabolites that can control phytopathogenic fungi in vitro, with the potential to adapt its effectiveness in the field. In the present study, leaf extracts of A. mexicana (hexane and methanol) were prepared, [...] Read more.

Argemone mexicana L. is considered a weed; however, it contains secondary metabolites that can control phytopathogenic fungi in vitro, with the potential to adapt its effectiveness in the field. In the present study, leaf extracts of A. mexicana (hexane and methanol) were prepared, and their chemical profiles were analyzed using gas chromatography–mass spectrometry (GC-MS). The in vitro antifungal activity of each extract was evaluated at different concentrations (500, 1000, 2000, 4000, and 8000 mg L⁻¹) against phytopathogens such as Monilinia fructicola, Colletotrichum gloeosporioides, Fusarium oxysporum, and Sclerotinia sclerotiorum. Based on their chemical profiles, 14 compounds were identified in the hexanic extract, and 11 compounds were identified in the methanolic extract. These compounds included those with antifungal activity, such as Benzene; 1.3-bis(1.1-dimethylethyl)-; pentanoic acid; 5-hydroxy-, 2,4-di-1-butylphenyl esters; 1,2,4-Triazol-4-amine; and N-(2-thienylmethyl). The hexanic extract demonstrated fungistatic activity on the four fungi tested, while the methanolic extract exhibited fungicidal activity against C. gloeosporioides and F. oxysporum. The results of the Probit analysis showed variations in the sensitivity of phytopathogenic fungi to the treatments evaluated. In M. fructicola, the hexane extract presented an EC₅₀ of 317,146 mg L⁻¹ and an EC₉₀ of 400,796 mg L⁻¹. For C. gloeosporioides, the EC₅₀ was 2676 mg L⁻¹ and the EC₉₀ was 888,177 mg L⁻¹, while in F. oxysporum an EC₅₀ of 34,274 mg L⁻¹ and an EC₉₀ of 1528 mg L⁻¹ were estimated. In the case of S. sclerotiorum, an EC₅₀ of 560 mg L⁻¹ and an EC₉₀ of 7776 mg L⁻¹ were obtained. Finally, for the commercial fungicide Captan^®, an EC₅₀ of 1.19 mg L⁻¹ and an EC₉₀ of 1.67 mg L⁻¹. These results suggest that extracts from A. mexicana could provide a natural alternative for the control of phytopathogenic fungi. Full article

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11 pages, 6634 KB

Open AccessCommunication

A Simple and Safe Protocol for Intra-Testicular Gene Delivery in Neonatal Mice Using a Convenient Isoflurane-Based Anesthesia System

by Kazunori Morohoshi, Miho Ohba, Masahiro Sato and Shingo Nakamura

BioTech 2025, 14(4), 81; https://doi.org/10.3390/biotech14040081 - 22 Oct 2025

Viewed by 163

Abstract

Newborn mice (up to 6 d after birth) are suitable for genetic manipulations, such as facial vein-mediated injection, owing to their hairless and thin skin. Their small body volumes also facilitate the rapid dissemination of injected solutions, supporting gene engineering-related experiments. However, anesthesia [...] Read more.

Newborn mice (up to 6 d after birth) are suitable for genetic manipulations, such as facial vein-mediated injection, owing to their hairless and thin skin. Their small body volumes also facilitate the rapid dissemination of injected solutions, supporting gene engineering-related experiments. However, anesthesia in newborns is challenging because of the potential risks associated with anesthetic agents. Isoflurane inhalation anesthesia is an option, although its effects on brain development remain under investigation. In this study, we established a reproducible protocol for delivering nucleic acids to juvenile mouse testes using a simple isoflurane-based anesthetic system prepared from common laboratory equipment. Using this system, nucleic acids were successfully delivered to juvenile mouse testes via intra-testicular injection, followed by in vivo electroporation. The present isoflurane-based method achieved >90% postoperative survival with normal maternal nursing observations. Gene delivery resulted in limited transfection of seminiferous tubules but efficient interstitial Leydig cell transfection. Thus, gene engineering in somatic and germ cells in neonatal mice will be facilitated using the anesthetic protocol established in this study. Full article

(This article belongs to the Section Biotechnology Regulation)

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25 pages, 5782 KB

Open AccessReview

Molecular Docking as a Key Driver of Biocontrol for Agri-Food Security

by María Isabel Iñiguez-Luna, Jorge David Cadena-Zamudio, Marco A. Ramírez-Mosqueda, José Luis Aguirre-Noyola, Daniel Alejandro Cadena-Zamudio, Jorge Cadena-Iñiguez and Alma Armenta-Medina

BioTech 2025, 14(4), 80; https://doi.org/10.3390/biotech14040080 - 14 Oct 2025

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Abstract

Molecular docking has emerged as a pivotal computational approach in agri-food research, offering a rapid and targeted means to discover bioactive molecules for crop protection and food safety. Its ability to predict and visualize interactions between natural or synthetic compounds and specific biological [...] Read more.

Molecular docking has emerged as a pivotal computational approach in agri-food research, offering a rapid and targeted means to discover bioactive molecules for crop protection and food safety. Its ability to predict and visualize interactions between natural or synthetic compounds and specific biological targets provides valuable opportunities to address urgent agricultural challenges, including climate change and the rise in resistant crop pathogens. By enabling the in silico screening of diverse chemical entities, this technique facilitates the identification of molecules with antimicrobial and antifungal properties, specifically designed to interact with critical enzymatic pathways in plant pathogens. Recent advancements, such as the integration of molecular dynamics simulations and artificial intelligence-enhanced scoring functions, have significantly improved docking accuracy by addressing limitations like protein flexibility and solvent effects. These technological improvements have accelerated the discovery of eco-friendly biopesticides and multifunctional nutraceutical agents. Promising developments include nanoparticle-based delivery systems that enhance the stability and efficacy of bioactive molecules. Despite its potential, molecular docking still faces challenges related to incomplete protein structures, variability in scoring algorithms, and limited experimental validation in agricultural contexts. This work highlights these limitations while outlining current trends and future prospects to guide its effective application in agri-food biotechnology. Full article

(This article belongs to the Topic From Plant to Pharmacology: Understanding the Metabolism of Natural Products)

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24 pages, 2566 KB

Open AccessReview

Valorization of Second Cheese Whey Through Microalgae-Based Treatments: Advantages, Limits, and Opportunities

by Gloria Sciuto, Nunziatina Russo, Cinzia L. Randazzo and Cinzia Caggia

BioTech 2025, 14(4), 79; https://doi.org/10.3390/biotech14040079 - 9 Oct 2025

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Abstract

The dairy sector produces considerable amounts of nutrient-rich effluents, which are frequently undervalued as simple by-products or waste. In particular, Second Cheese Whey (SCW), also known as scotta, exhausted whey, or deproteinized whey, represents the liquid fraction from ricotta cheese production. Despite its [...] Read more.

The dairy sector produces considerable amounts of nutrient-rich effluents, which are frequently undervalued as simple by-products or waste. In particular, Second Cheese Whey (SCW), also known as scotta, exhausted whey, or deproteinized whey, represents the liquid fraction from ricotta cheese production. Despite its abundance and high organic and saline content, SCW is often improperly discharged into terrestrial and aquatic ecosystems, causing both environmental impact and resource waste. The available purification methods are expensive for dairy companies, and, at best, SCW is reused as feed or fertilizer. In recent years, increasing awareness of sustainability and circular economy principles has increased interest in the valorization of SCW. Biological treatment of SCW using microalgae represents an attractive strategy, as it simultaneously reduces the organic load and converts waste into algal biomass. This biomass can be further valorized as a source of proteins, pigments, and bioactive compounds with industrial relevance, supporting applications in food, nutraceuticals, biofuels, and cosmetics. This review, starting from analyzing the characteristics, production volumes, and environmental issues associated with SCW, focused on the potential of microalgae application for their valorization. In addition, the broader regulatory and sustainability aspects related to biomass utilization and treated SCW are considered, highlighting both the promises and limitations of microalgae-based strategies by integrating technological prospects with policy considerations. Full article

(This article belongs to the Special Issue A Sustainable Approach to Macroalgae and Microalgae: From Cultivation to Compound Recovery and Biotechnological Applications)

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24 pages, 13022 KB

Open AccessArticle

Development of PCR Methods for Detecting Wheat and Maize Allergens in Food

by Tata Ninidze, Tamar Koberidze, Kakha Bitskinashvili, Tamara Kutateladze, Boris Vishnepolsky and Nelly Datukishvili

BioTech 2025, 14(4), 78; https://doi.org/10.3390/biotech14040078 - 1 Oct 2025

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Abstract

The detection of allergens is essential for ensuring food safety, protecting public health, and providing accurate information to consumers. Wheat (Triticum aestivum L.) and maize (Zea mays L.) are recognized as important food allergens. In this study, novel PCR methods were [...] Read more.

The detection of allergens is essential for ensuring food safety, protecting public health, and providing accurate information to consumers. Wheat (Triticum aestivum L.) and maize (Zea mays L.) are recognized as important food allergens. In this study, novel PCR methods were developed for the reliable detection of wheat and maize allergens, including wheat high-molecular-weight glutenin subunit (HMW-GS) and low-molecular-weight glutenin subunit (LMW-GS), as well as three maize allergens, namely, Zea m 14, Zea m 8, and zein. Wheat and maize genomic DNA, as well as allergen genes, were examined during 60 min of baking at 180 °C and 220 °C. Agarose gel electrophoresis revealed degradation of genomic DNA and amplified PCR fragments in correlation with increasing baking temperature and time. For each target gene, the best primers were identified that could detect HMW-GS and LMW-GS genes in wheat samples and Zea m 14, Zea m 8, and zein genes in maize samples after baking at 220 °C for 60 min and 40 min, respectively. The results indicate that these PCR methods can be used for the reliable and sensitive detection of wheat and maize allergens in processed foods. Full article

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

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15 pages, 3351 KB

Open AccessArticle

Biotic Elicitor-Driven Enhancement of In Vitro Micropropagation and Organogenesis in Solanum tuberosum L. cv. Fianna

by Mario James-Forest, Ma del Carmen Ojeda-Zacarías, Alhagie K. Cham, Héctor Lozoya-Saldaña, Rigoberto E. Vázquez-Alvarado, Emilio Olivares-Sáenz and Alejandro Ibarra-López

BioTech 2025, 14(4), 77; https://doi.org/10.3390/biotech14040077 - 24 Sep 2025

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Abstract

This study evaluates the impact of biotic elicitors and hormone regimes on the in vitro establishment, shoot multiplication, and organogenesis of Solanum tuberosum L. cv. Fianna under controlled laboratory conditions. Explants derived from pre-treated tubers were cultured on Murashige and Skoog (MS) medium [...] Read more.

This study evaluates the impact of biotic elicitors and hormone regimes on the in vitro establishment, shoot multiplication, and organogenesis of Solanum tuberosum L. cv. Fianna under controlled laboratory conditions. Explants derived from pre-treated tubers were cultured on Murashige and Skoog (MS) medium supplemented with vitamins and varying concentrations of growth regulators or elicitors. Aseptic establishment achieved a high success rate (~95%) using a 6% sodium hypochlorite disinfection protocol. Multiplication was significantly enhanced with a combination of 0.2 mg L⁻¹ naphthaleneacetic acid (NAA) and 0.5–1.0 mg L⁻¹ benzylaminopurine (BAP), producing the greatest number and length of shoots and roots. Direct organogenesis was stimulated by bio-elicitors Activane^®, Micobiol^®, and Stemicol^® in (MS) basal medium at mid-level concentrations (0.5 g or mL L⁻¹), improving shoot number, elongation, and root development. Activane^®, Micobiol^®, and Stemicol^® are commercial elicitors that stimulate plant defense pathways and morphogenesis through salicylic acid, microbial, and jasmonic acid signaling mechanisms, respectively. Indirect organogenesis showed significantly higher callus proliferation in Stemicol^® and Micobiol^® treatments compared to the control medium, resulting in the highest fresh weight, diameter, and friability of callus. The results demonstrate the potential of biotic elicitors as alternatives or enhancers to traditional plant growth regulators in potato tissue culture, supporting more efficient and cost-effective micropropagation strategies. Full article

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

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15 pages, 1133 KB

Open AccessArticle

Evaluating Scale-Up Cultivation Modes for Aspergillus oryzae Biomass Production Using VFA-Rich Effluents from Agro-Industrial Residues

by Taner Sar, Clarisse Uwineza, Mohammad J. Taherzadeh and Amir Mahboubi

BioTech 2025, 14(4), 76; https://doi.org/10.3390/biotech14040076 - 24 Sep 2025

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Abstract

Organic-waste-derived volatile fatty acids (VFAs) are promising substrates for fungal biomass cultivation, offering a nutrient-rich medium capable of meeting microbial growth requirements. However, the growth and biomass productivity are highly influenced by the VFAs’ composition and mode of operation. This study investigated the [...] Read more.

Organic-waste-derived volatile fatty acids (VFAs) are promising substrates for fungal biomass cultivation, offering a nutrient-rich medium capable of meeting microbial growth requirements. However, the growth and biomass productivity are highly influenced by the VFAs’ composition and mode of operation. This study investigated the cultivation of Aspergillus oryzae fungal biomass using agro-industrial-derived VFA effluent, employing repeated-batch and fed-batch (stepwise and continuous-feeding) cultivation modes to evaluate fungal growth and biomass composition. The highest dry biomass yield of 0.41 dry biomass/gVFAs_fed (g/g) was achieved in fed-batch mode with continuous feeding, where the biomass exhibited pellet morphology, facilitating ease of harvesting. The crude protein content varied according to the cultivation strategy, reaching 45–53% in continuous-feeding fed-batch mode, while it was 34–42% in stepwise fed-batch mode. Additionally, the fungal biomass contained significant levels of essential macronutrients and trace elements, including Mg, Ca, K, Mn, and Fe, which are crucial if the biomass is intended to be used in animal feed formulations. This study highlights the effects of cultivation modes on biomass composition and the potential of VFA-derived fungal biomass as a sustainable feed ingredient. Full article

(This article belongs to the Special Issue Volatile Fatty Acids (VFAs) and Their Potential in Waste-Based Biorefineries)

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BioTech, Volume 14, Issue 4 (December 2025) – 11 articles

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