BioTech

16 pages, 2825 KiB

Open AccessArticle

Bioremediation Potential of a Non-Axenic Cyanobacterium Synechococcus sp. for Municipal Wastewater Treatment in the Peruvian Amazon: Growth Kinetics, Ammonium Removal, and Biochemical Characterization Within a Circular Bioeconomy Framework

by Remy G. Cabezudo, Juan C. Castro, Carlos G. Castro, Hicler N. Rodriguez, Gabriela L. García, Paul M. Vizcarra, Carmen Ruiz-Huamán and Marianela Cobos

BioTech 2025, 14(2), 36; https://doi.org/10.3390/biotech14020036 - 13 May 2025

Abstract

Effective wastewater management is critical for mitigating environmental and health impacts in ecologically sensitive regions like the Peruvian Amazon, where rapid urbanization has led to increased discharge of nutrient-rich effluents into freshwater systems. Conventional treatment methods often fail to address nutrient imbalances while [...] Read more.

Effective wastewater management is critical for mitigating environmental and health impacts in ecologically sensitive regions like the Peruvian Amazon, where rapid urbanization has led to increased discharge of nutrient-rich effluents into freshwater systems. Conventional treatment methods often fail to address nutrient imbalances while generating secondary pollutants. This study aims to evaluate the bioremediation potential of a non-axenic cyanobacterium, Synechococcus sp., isolated from the Amazon Basin, for municipal wastewater treatment within a circular bioeconomy framework. The strain was cultivated in different concentrations of municipal wastewater (25%, 50%, 75%, 100%) from Moronacocha Lake in the Peruvian Amazon to assess growth kinetics, ammonium removal efficiency, and biochemical composition. The cyanobacterium exhibited optimal performance in 25% wastewater, achieving the highest specific growth rate (22.8 × 10⁻² μ·day⁻¹) and biomass increase (393.2%), exceeding even the standard BG-11 medium. This treatment also demonstrated exceptional ammonium removal efficiency (95.4%) and enhanced phycocyanin production (33.6 μg/mg, 56% higher than the control). As wastewater concentration increased, both growth parameters and removal efficiency progressively declined. Biochemical analysis revealed that higher wastewater concentrations resulted in decreased protein content and increased lipid accumulation in the biomass. These findings demonstrate the dual potential of Synechococcus sp. for effective wastewater remediation and production of valuable biomass with modifiable biochemical characteristics, offering a sustainable approach for wastewater management in the Peruvian Amazon region. Full article

(This article belongs to the Topic Sustainable Water Purification Technologies for Multiple Applications)

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32 pages, 2714 KiB

Open AccessArticle

Comparative Potential of Chitinase and Chitosanase from the Strain Bacillus thuringiensis B-387 for the Production of Antifungal Chitosan Oligomers

by Gleb Aktuganov, Alexander Lobov, Nailya Galimzianova, Elena Gilvanova, Lyudmila Kuzmina, Polina Milman, Alena Ryabova, Alexander Melentiev, Sergey Chetverikov, Sergey Starikov and Sergey Lopatin

BioTech 2025, 14(2), 35; https://doi.org/10.3390/biotech14020035 - 8 May 2025

Abstract

The depolymerization of chitosan using chitinolytic enzymes is one of the most promising approaches for the production of bioactive soluble chitooligosaccharides (COS) due to its high specificity, environmental safety, mild reaction conditions, and potential for development. However, the comparative efficacy of bacterial chitinases [...] Read more.

The depolymerization of chitosan using chitinolytic enzymes is one of the most promising approaches for the production of bioactive soluble chitooligosaccharides (COS) due to its high specificity, environmental safety, mild reaction conditions, and potential for development. However, the comparative efficacy of bacterial chitinases and chitosanases in terms of yield, solubility, and antimicrobial activity of produced COS remains understudied. In this work, chitinase (73 kDa) and chitosanase (40 kDa) from the strain Bacillus thuringiensis B-387 (Bt-387) were purified using various chromatographic techniques and compared by their action on chitosan (DD 85%). The molecular mass and structure of generated COS was determined using TLC, LC-ESI-MS, HP-SEC, and C¹³-NMR techniques. Chitosanase converted the polymer more rapidly to short COS (GlcN₂-GlcN₄), than chitinase, and was more specific in its action on mixed bonds between GlcN and GlcNAc. Chitosanase needed a noticeably shorter incubation time and enzyme–substrate ratio than chitinase for production of larger oligomeric molecules (Mw 2.4–66.5 and 15.4–77.7 kDa, respectively) during controlled depolymerization of chitosan. Moreover, chitosanase-generated oligomers demonstrate better solubility and a higher antifungal activity in vitro against the tested plant pathogenic fungi. These features, as well as the high enzyme production and its simplified purification protocol, make chitosanase B-387 more suitable for the production of antifungal chitooligomers than chitinase. Full article

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17 pages, 2839 KiB

Open AccessArticle

Combined Effect of Spent Mushroom Substrate and Agro-Industrial Residues on Pleurotus columbinus Production and Intra-Cellular Polysaccharide Synthesis

by Marianna Dedousi, Chrysavgi Gardeli, Seraphim Papanikolaou and Panagiota Diamantopoulou

BioTech 2025, 14(2), 34; https://doi.org/10.3390/biotech14020034 - 2 May 2025

Abstract

Spent mushroom substrate (SMS), spent coffee grounds from espresso production (SCG), faba bean harvest residues (FBR), pistachio shells (PS) wheat straw (WS) (control) agro-industrial waste were combined in different ratios, with or without supplements (wheat bran, soybean flour), to create novel substrates for [...] Read more.

Spent mushroom substrate (SMS), spent coffee grounds from espresso production (SCG), faba bean harvest residues (FBR), pistachio shells (PS) wheat straw (WS) (control) agro-industrial waste were combined in different ratios, with or without supplements (wheat bran, soybean flour), to create novel substrates for Pleurotus columbinus growth. The impact of the substrates on the mycelial growth rate (Kr), biomass production, laccase, total cellulases and carbohydrate synthesis, along with the C and N consumption by P. columbinus, were examined in fully colonized substrates. The incubation period, earliness and biological efficiency (B.E.) (%) were also determined. Then, the intracellular polysaccharide (ICP) contents of the P. columbinus produced mushrooms were evaluated in the most promising substrates. P. columbinus was grown successfully in a wide range of C/N ratios of substrates and the fastest Kr (7.6 mm/d) was detected on the 70 SMS-30 FBR, without supplements, whereas substrates consisting of SCG enhanced biomass production (700.0–803.7 mg/g d.w.). SMS and PS or SCG led to the shortest incubation and earliness period of P. columbinus. The C content was reduced and the N content was substantially increased in all the colonized substrates. The 70 SMS-30 FBR and 80 SMS considerably enhanced the laccase production (up to 59,933.4 U/g d.w.) and substrates consisting of PS promoted total cellulases activities. Greater amounts of carbohydrates (3.8–17.4 mg/g d.w.) than that in the control were recorded for all the substrates. The combination of SMS and SCG or WS led to the highest B.E. values (59.3–87.1%) and ICP amounts (34.7–45.9%, w/w), regardless of the supplement addition. These findings support the effective utilization of agro-industrial waste in P. columbinus cultivation, producing high-value-added compounds and supporting mushroom growth. Full article

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27 pages, 10074 KiB

Open AccessArticle

Innovations in Proteomic Technologies and Artificial Neural Networks: Unlocking Milk Origin Identification

by Achilleas Karamoutsios, Emmanouil D. Oikonomou, Chrysoula (Chrysa) Voidarou, Lampros Hatzizisis, Konstantina Fotou, Konstantina Nikolaou, Evangelia Gouva, Evangelia Gkiza, Nikolaos Giannakeas, Ioannis Skoufos and Athina Tzora

BioTech 2025, 14(2), 33; https://doi.org/10.3390/biotech14020033 - 28 Apr 2025

Abstract

Milk’s biological origin determination, including its adulteration and authenticity, presents serious limitations, highlighting the need for innovative advanced solutions. The utilisation of proteomic technologies combined with personalised algorithms creates great potential for a more comprehensive approach to analysing milk samples effectively. The current [...] Read more.

Milk’s biological origin determination, including its adulteration and authenticity, presents serious limitations, highlighting the need for innovative advanced solutions. The utilisation of proteomic technologies combined with personalised algorithms creates great potential for a more comprehensive approach to analysing milk samples effectively. The current study presents an innovative approach utilising proteomics and neural networks to classify and distinguish bovine, ovine and caprine milk samples by employing advanced machine learning techniques; we developed a precise and reliable model capable of distinguishing the unique mass spectral signatures associated with each species. Our dataset includes a diverse range of mass spectra collected from milk samples after MALDI-TOF MS (Matrix-assisted laser desorption/ionization-time of flight mass spectrometry) analysis, which were used to train, validate, and test the neural network model. The results indicate a high level of accuracy in species identification, underscoring the model’s potential applications in dairy product authentication, quality assurance, and food safety. The current research offers a significant contribution to agricultural science, providing a cutting-edge method for species-specific classification through mass spectrometry. The dataset comprises 648, 1554, and 2392 spectra, represented by 16,018, 38,394, and 55,055 eight-dimensional vectors from bovine, caprine, and ovine milk, respectively. Full article

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21 pages, 2688 KiB

Open AccessArticle

Production of Multifunctional Hydrolysates from the Lupinus mutabilis Protein Using a Micrococcus sp. PC7 Protease

by Keyla Sofía Llontop-Bernabé, Arturo Intiquilla, Carlos Ramirez-Veliz, Marco Santos, Karim Jiménez-Aliaga, Amparo Iris Zavaleta, Samuel Paterson and Blanca Hernández-Ledesma

BioTech 2025, 14(2), 32; https://doi.org/10.3390/biotech14020032 - 27 Apr 2025

Abstract

The growing demand for functional foods has driven the search for bioactive compounds derived from plant proteins. Lupinus mutabilis “Tarwi”, a legume native to the Peruvian Andes, stands out for its high protein content and potential as a source of bioactive peptides (BPs). [...] Read more.

The growing demand for functional foods has driven the search for bioactive compounds derived from plant proteins. Lupinus mutabilis “Tarwi”, a legume native to the Peruvian Andes, stands out for its high protein content and potential as a source of bioactive peptides (BPs). In this study, the functionality of the proteins contained in the albumin fraction (AF) isolated by tangential ultrafiltration (TFF) was investigated by using the OmicsBox software. The identified proteins were functionally classified into three groups: cellular component (35.57%), molecular function (33.45%), and biological process (30.97%). The isolated AF was hydrolysed with the native protease PC7 (HAP), optimizing the E/S ratio and time parameters. Additionally, sequential hydrolysis of the PC7 protease and alcalase (HAPA) was performed. In vitro multifunctionality assays, HAP and HAPA demonstrated the ability to scavenge radicals (ABTS and ORAC) and inhibit angiotensin-converting enzyme (ACE)-I and dipeptidyl peptidase IV (DPP-IV). The findings of this study highlight the potential of L. mutabilis albumin hydrolysate as a multifunctional ingredient for functional foods aimed at managing chronic conditions associated with oxidative stress, hypertension, and/or metabolic disorders. Full article

(This article belongs to the Special Issue Natural Antioxidants: Determination in Food and Nutraceuticals and Implications on Human Health)

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14 pages, 5306 KiB

Open AccessArticle

Vaginal Microbiota Patterns Associated with Yeast Infection in Mexican Women, a Pilot Study

by Janet Pineda-Díaz, Carolina Miranda-Brito, Carmen Josefina Juárez-Castelán, Alberto Piña-Escobedo, Noemí del Socorro Lázaro-Pérez, Alejandra de la Cruz-Munguía, Daniela Ramírez-Sánchez, Yuliana Gómez-Meraz, Juan Manuel Vélez-Ixta and Jaime García-Mena

BioTech 2025, 14(2), 31; https://doi.org/10.3390/biotech14020031 - 26 Apr 2025

Abstract

Recurrent vulvovaginal candidiasis (RVVC) is a common condition that affects women of reproductive age. The etiology of RVVC remains largely unknown, but it is believed to be associated with changes in vaginal microbiota composition. This study investigates the vaginal microbiota in 57 women [...] Read more.

Recurrent vulvovaginal candidiasis (RVVC) is a common condition that affects women of reproductive age. The etiology of RVVC remains largely unknown, but it is believed to be associated with changes in vaginal microbiota composition. This study investigates the vaginal microbiota in 57 women with RVVC and 38 healthy controls. Bacterial DNA was analyzed using high-throughput 16S rRNA gene sequencing, and Candida and Saccharomyces species were determined by PCR. RVVC cases had a higher prevalence of Nakaseomyses glabratus (former Candida glabrata) compared to controls. Alpha diversity metrics were similar between groups, but beta diversity analysis revealed significant differences in vaginal microbiota composition. The Firmicutes abundance was altered in RVVC cases, with genus Bifidobacterium and phylum Actinobacteriota being more abundant than in the controls. At the genus level, Lactobacillus dominated controls using antibiotics, while Bifidobacterium was higher in cases with no antibiotic intake. Our study provides evidence that Nakaseomyses glabratus (former Candida glabrata) is a significant pathogen in RVVC, while Candida albicans was more prevalent in healthy women. The vaginal microbiota composition differs significantly between the two groups, with distinct patterns of bacterial abundance and changes in Firmicutes abundance. Full article

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13 pages, 2641 KiB

Open AccessArticle

Characterization and Biotechnology of Three New Strains of Basidial Fungi as Promising Sources of Biologically Active Substances

by Maria Alexandrovna Sysoeva, Ilyuza Shamilevna Prozorova, Elena Vladislavovna Sysoeva, Tatyana Vladimirovna Grigoryeva and Ruzilya Kamilevna Ismagilova

BioTech 2025, 14(2), 30; https://doi.org/10.3390/biotech14020030 - 25 Apr 2025

Abstract

The study of new strains of basidiomycetes as sources of biologically active substances is a promising direction in modern biotechnology. This work aims to isolate new cultures of the fungi Daedaleopsis tricolor, Pycnoporellus fulgens and Trichaptum abietinum from natural fruiting bodies and [...] Read more.

The study of new strains of basidiomycetes as sources of biologically active substances is a promising direction in modern biotechnology. This work aims to isolate new cultures of the fungi Daedaleopsis tricolor, Pycnoporellus fulgens and Trichaptum abietinum from natural fruiting bodies and to improve their growth conditions on solid nutrient media. The identification of fungi was performed based on their morphological features and using the Sanger sequencing method. Cultivation was carried out by placing inoculum in the middle of a Petri dish and at the edge, which provided a more comprehensive definition of the characteristics of colonies and fungus hyphae. New strains were registered in Genbank Overview. The optimal cultivation temperature was 27 °C for all studied strains. The highest radial growth was observed on synthetic medium for D. tricolor (5.26 mm/day) and T. abietinum (7.5 mm/day), and on synthetic medium with lignin for P. fulgens (2.98 mm/day). The biomass amount of D. tricolor KS11 was 133.25 mg at 9 days of cultivation, that of P. fulgens KS12 was 86.73 mg at 16 days, and that of T. abietinum KS10 was 227.33 mg at 6 days. New strains of fungi can be used to obtain biologically active substances for the food, pharmaceutical and cosmetic industries. Full article

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

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12 pages, 3005 KiB

Open AccessArticle

Direct Shoot Regeneration from the Finger Millet’s In Vitro-Derived Shoot Apex and Genetic Fidelity Study with ISSR Markers

by Theivanayagam Maharajan, Veeramuthu Duraipandiyan and Thumadath Palayullaparambil Ajeesh Krishna

BioTech 2025, 14(2), 29; https://doi.org/10.3390/biotech14020029 - 18 Apr 2025

Abstract

Globally, people are cultivating finger millet, an important cereal, to improve food availability and health benefits for humans. However, the biotechnological research on this millet is limited and insufficient in this field. The primary focus of this study is to optimize an efficient [...] Read more.

Globally, people are cultivating finger millet, an important cereal, to improve food availability and health benefits for humans. However, the biotechnological research on this millet is limited and insufficient in this field. The primary focus of this study is to optimize an efficient regenerated protocol for initiating further plant transformation studies, using the shoot apex as an explant and various growth regulators. For example, three cytokinins (BAP, TDZ, and Kin) at different concentrations were used to induce multiple shoots of finger millet. Among these, TDZ (4.5 µM) provided the maximum number (17.3) of shoots as compared to BAP and Kin. IBA (2.46 µM), along with MS medium, was used for the induction of roots, where 5.6 roots were produced in an individual shoot and the length of the root was longer with a size of 8.2 cm after two weeks of incubation. The clonal fidelity of the in vitro regenerated plantlets of finger millet was confirmed by ISSR primers. Overall, the present work developed a robust and reliable procedure for the establishment of efficient and reproducible regeneration through the shoot apex that will be useful for the genetic improvement of this crop. The genetic enhancement of these millets as well as the successful creation of transgenic plant varieties modified for resistance to biotic and abiotic challenges in the near future would be aided by this study. Full article

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

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26 pages, 1171 KiB

Open AccessReview

Matrix Metalloproteinases in Glioma: Drivers of Invasion and Therapeutic Targets

by Ella E. Aitchison, Alexandra M. Dimesa and Alireza Shoari

BioTech 2025, 14(2), 28; https://doi.org/10.3390/biotech14020028 - 15 Apr 2025

Abstract

Matrix metalloproteinases (MMPs) are a family of zinc-dependent proteolytic enzymes that are crucial for the remodeling of the extracellular matrix, a process that is often co-opted by cancers, including brain tumors, to facilitate growth, invasion, and metastasis. In gliomas, MMPs contribute to a [...] Read more.

Matrix metalloproteinases (MMPs) are a family of zinc-dependent proteolytic enzymes that are crucial for the remodeling of the extracellular matrix, a process that is often co-opted by cancers, including brain tumors, to facilitate growth, invasion, and metastasis. In gliomas, MMPs contribute to a complex interplay involving tumor proliferation, angiogenesis, and immune modulation, thereby influencing tumor progression and patient prognosis. This review provides a comprehensive analysis of the roles of various MMPs in different types of gliomas, from highly malignant gliomas to metastatic lesions. Emphasis is placed on how the dysregulation of MMPs impacts tumor behavior, the association between specific MMPs and the tumor grade, and their potential as biomarkers for diagnosis and prognosis. Additionally, the current therapeutic approaches targeting MMP activity are discussed, exploring both their challenges and future potential. By synthesizing recent findings, this paper aims to clarify the broad significance of MMPs in gliomas and propose avenues for translational research that could enhance treatment strategies and clinical outcomes. Full article

(This article belongs to the Section Medical Biotechnology)

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18 pages, 2479 KiB

Open AccessArticle

AsNAC Genes: Response to High Mercury Concentrations in Allium sativum Seed Clove

by Brenda Mendoza-Almanza, María de la Luz Guerrero-González, Marcos Loredo-Tovias, María Elena García-Arreola, Catarina Loredo-Osti, Erika Padilla-Ortega and Pablo Delgado-Sánchez

BioTech 2025, 14(2), 27; https://doi.org/10.3390/biotech14020027 - 8 Apr 2025

Abstract

Heavy metal contamination in soils is a growing concern due to anthropogenic activities, and Allium sativum (garlic) has shown tolerance to mercury pollution. We analyzed the physiological and molecular responses of garlic cloves exposed to HgCl₂ at 0, 5000, 23,000, and 46,000 [...] Read more.

Heavy metal contamination in soils is a growing concern due to anthropogenic activities, and Allium sativum (garlic) has shown tolerance to mercury pollution. We analyzed the physiological and molecular responses of garlic cloves exposed to HgCl₂ at 0, 5000, 23,000, and 46,000 mg/kg for 2, 3, and 4 h. The germination percentage was lower than 46,000 mg/kg Hg for 4 h. We also analyzed the expression levels of NAC transcription factors and found that AsNAC11 had higher expression at 46,000 mg/kg at 2 h; AsNAC17 was underexpressed and the maximum was at 2 h at 23,000 mg/kg. AsNAC20 had the highest expression (30 times more than the control) at 3 and 4 h with 23,000 mg/Kg. AsNAC27 showed the highest expression at 3 h with 23,000 mg/kg. The tissues exhibited a maximum Hg bioconcentration factor of 0.037 at 23,000 mg/kg, indicating moderate mercury absorption. However, at a concentration of 46,000 mg/kg, the BCF decreased to 0.023. Our in-silico analysis revealed that the analyzed AsNACs are associated with various abiotic stress responses. This study provides valuable insights into genes that could be utilized for genetic improvement to enhance crop resistance to mercury soil contamination. Full article

(This article belongs to the Section Environmental Biotechnology)

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13 pages, 1406 KiB

Open AccessReview

A Review of Classical and Rising Approaches the Extraction and Utilization of Marine Collagen

by Cesia Deyanira Gutierrez-Canul, Luis Alfonso Can-Herrera, Emmanuel de Jesús Ramírez-Rivera, Witoon Prinyawiwatkul, Enrique Sauri-Duch, Victor Manuel Moo-Huchin and Emanuel Hernández-Núñez

BioTech 2025, 14(2), 26; https://doi.org/10.3390/biotech14020026 - 3 Apr 2025

Abstract

This comprehensive review explores the extraction and utilization of marine collagen, a sustainable alternative to traditional mammalian sources. The review covers conventional extraction methods like acid and pepsin solubilization, highlighting their limitations and contributing to the search for improved efficiency and sustainability. It [...] Read more.

This comprehensive review explores the extraction and utilization of marine collagen, a sustainable alternative to traditional mammalian sources. The review covers conventional extraction methods like acid and pepsin solubilization, highlighting their limitations and contributing to the search for improved efficiency and sustainability. It also delves into innovative extraction technologies, such as ultrasound-assisted extraction, deep eutectic solvents, and supercritical carbon dioxide, showing their potential to revolutionize the field. The significance of collagen hydrolysis in generating bioactive peptides with diverse functionalities is also discussed, emphasizing their potential applications in various sectors. By providing an analysis of marine collagen extraction and its implications, this review presents a perspective for leveraging this valuable bioresource, promoting a circular economy, and satisfying the increasing demand for high-quality collagen in diverse industries. Full article

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17 pages, 1547 KiB

Open AccessArticle

Green Biocatalysis of Xylitol Monoferulate: Candida antarctica Lipase B-Mediated Synthesis and Characterization of Novel Bifunctional Prodrug

by Federico Zappaterra, Francesco Presini, Domenico Meola, Chaimae Chaibi, Simona Aprile, Lindomar Alberto Lerin and Pier Paolo Giovannini

BioTech 2025, 14(2), 25; https://doi.org/10.3390/biotech14020025 - 2 Apr 2025

Abstract

Natural compounds with significant bioactive properties can be found in abundance within biomasses. Especially prominent for their anti-inflammatory, neuroprotective, antibacterial, and antioxidant activities are cinnamic acid derivatives (CAs). Ferulic acid (FA), a widely studied phenylpropanoid, exhibits a broad range of therapeutic and nutraceutical [...] Read more.

Natural compounds with significant bioactive properties can be found in abundance within biomasses. Especially prominent for their anti-inflammatory, neuroprotective, antibacterial, and antioxidant activities are cinnamic acid derivatives (CAs). Ferulic acid (FA), a widely studied phenylpropanoid, exhibits a broad range of therapeutic and nutraceutical applications, demonstrating antidiabetic, anticancer, antimicrobial, and hepato- and neuroprotective activities. This research investigates the green enzymatic synthesis of innovative and potentially bifunctional prodrug derivatives of FA, designed to enhance solubility and stability profiles. Selective esterification was employed to conjugate FA with xylitol, a biobased polyol recognized for its bioactive antioxidant properties and safety profile. Furthermore, by exploiting t-amyl alcohol as a green solvent, the enzymatic synthesis of the derivative was optimized for reaction parameters including temperature, reaction time, enzyme concentration, and molar ratio. The synthesized derivative, xylitol monoferulate (XMF), represents a novel contribution to the literature. The comprehensive characterization of this compound was achieved using advanced spectroscopic methods, including ¹H-NMR, ¹³C-NMR, COSY, HSQC, and HMBC. This study represents a significant advancement in the enzymatic synthesis of high-value biobased derivatives, demonstrating increased biological activities and setting the stage for future applications in green chemistry and the sustainable production of bioactive compounds. Full article

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

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23 pages, 581 KiB

Open AccessArticle

Screening of Non-Conventional Yeasts on Low-Cost Carbon Sources and Valorization of Mizithra Secondary Cheese Whey for Metabolite Production

by Gabriel Vasilakis, Rezart Tefa, Antonios Georgoulakis, Dimitris Karayannis, Ioannis Politis and Seraphim Papanikolaou

BioTech 2025, 14(2), 24; https://doi.org/10.3390/biotech14020024 - 1 Apr 2025

Abstract

The production of microbial metabolites such as (exo)polysaccharides, lipids, or mannitol through the cultivation of microorganisms on sustainable, low-cost carbon sources is of high interest within the framework of a circular economy. In the current study, two non-extensively studied, non-conventional yeast strains, namely, [...] Read more.

The production of microbial metabolites such as (exo)polysaccharides, lipids, or mannitol through the cultivation of microorganisms on sustainable, low-cost carbon sources is of high interest within the framework of a circular economy. In the current study, two non-extensively studied, non-conventional yeast strains, namely, Cutaneotrichosporon curvatus NRRL YB-775 and Papiliotrema laurentii NRRL Y-3594, were evaluated for their capability to grow on semi-defined lactose-, glycerol-, or glucose-based substrates and produce value-added metabolites. Three different nitrogen-to-carbon ratios (i.e., 20, 80, 160 mol/mol) were tested in shake-flask batch experiments. Pretreated secondary cheese whey (SCW) was used for fed-batch bioreactor cultivation of P. laurentii NRRL Y-3594, under nitrogen limitation. Based on the screening results, both strains can grow on low-cost substrates, yielding high concentrations of microbial biomass (>20 g/L) under nitrogen-excess conditions, with polysaccharides comprising the predominant component (>40%, w/w, of dry biomass). Glucose- and glycerol-based cultures of C. curvatus promote the secretion of mannitol (13.0 g/L in the case of glucose, under nitrogen-limited conditions). The lipids (maximum 2.2 g/L) produced by both strains were rich in oleic acid (≥40%, w/w) and could potentially be utilized to produce second-generation biodiesel. SCW was nutritionally sufficient to grow P. laurentii strain, resulting in exopolysaccharides secretion (25.6 g/L), along with dry biomass (37.9 g/L) and lipid (4.6 g/L) production. Full article

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

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20 pages, 1727 KiB

Open AccessReview

Developments in Extracellular Matrix-Based Angiogenesis Therapy for Ischemic Heart Disease: A Review of Current Strategies, Methodologies and Future Directions

by Jad Hamze, Mark Broadwin, Christopher Stone, Kelsey C. Muir, Frank W. Sellke and M. Ruhul Abid

BioTech 2025, 14(1), 23; https://doi.org/10.3390/biotech14010023 - 19 Mar 2025

Abstract

Ischemic heart disease (IHD) is the leading cause of mortality worldwide, underscoring the urgent need for innovative therapeutic strategies. The cardiac extracellular matrix (ECM) undergoes extreme transformations during IHD, adversely influencing the heart’s structure, mechanics, and cellular signaling. Researchers investigating the regenerative capacity [...] Read more.

Ischemic heart disease (IHD) is the leading cause of mortality worldwide, underscoring the urgent need for innovative therapeutic strategies. The cardiac extracellular matrix (ECM) undergoes extreme transformations during IHD, adversely influencing the heart’s structure, mechanics, and cellular signaling. Researchers investigating the regenerative capacity of the diseased heart have turned their attention to exploring the modulation of ECM to improve therapeutic outcomes. In this review, we thoroughly examine the current state of knowledge regarding the cardiac ECM and its therapeutic potential in the ischemic myocardium. We begin by providing an overview of the fundamentals of cardiac ECM, focusing on the structural, functional, and regulatory mechanisms that drive its modulation. Subsequently, we examine the ECM’s interactions within both chronically ischemic and acutely infarcted myocardium, emphasizing key ECM components and their roles in modulating angiogenesis. Finally, we discuss recent ECM-based approaches in biomedical engineering, focusing on different types of scaffolds as delivery tools and their compositions, and conclude with future directions for therapeutic research. By harnessing the potential of these emerging ECM-based therapies, we aim to contribute to the development of novel therapeutic modalities for IHD. Full article

(This article belongs to the Section Medical Biotechnology)

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16 pages, 1473 KiB

Open AccessArticle

Identifying Antimicrobial Agents from Chlorella sorokiniana: A Biotechnological Approach Utilizing Eco-Friendly Extraction and Characterization Methods

by Elia Lio, Martina Dramis, Gianluca Ottolina and Francesco Secundo

BioTech 2025, 14(1), 22; https://doi.org/10.3390/biotech14010022 - 18 Mar 2025

Abstract

Natural compounds are increasingly favored over synthetic ones for their lower environmental impact. However, extraction and characterization processes typically rely on harsh conditions and conventional solvents, which are unsustainable and cause pollution. This study aimed to develop an eco-friendly extraction method to isolate [...] Read more.

Natural compounds are increasingly favored over synthetic ones for their lower environmental impact. However, extraction and characterization processes typically rely on harsh conditions and conventional solvents, which are unsustainable and cause pollution. This study aimed to develop an eco-friendly extraction method to isolate and evaluate the antimicrobial properties of bioactive compounds from Chlorella sorokiniana. Using dimethyl carbonate (DMC), methoxycyclopentane (CPME), and butan-2-one (MEK) as green solvents alongside chloroform as a non-green reference solvent, on both untreated and sodium hydroxide pre-treated microalgae biomass, extract yields of up to 182 ± 27 mg/g DW were obtained using MEK. Extracts from untreated microalgae biomass exhibited lower MIC values compared to those obtained with the same solvent from pre-treated biomass, when tested as antimicrobial agents against Escherichia coli, Bacillus megaterium, and Bacillus subtilis. The lowest MIC value (4.89 ± 0.05 µg/mL) was observed against E. coli using the extract from the untreated microalgae biomass with CPME, which was comparable to the vancomycin control (1.55 ± 0.03 µg/mL). Principal component analysis highlighted correlations between GC-MS-identified compounds and antimicrobial activity. ANOVA and post hoc tests (p < 0.05) confirmed solvent choice, and pre-treatment influenced yield and bioactivity. The results underscore green solvents as sustainable alternatives for extracting bioactive compounds from autotrophic microalgae. Full article

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

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17 pages, 825 KiB

Open AccessArticle

Genomic Characterization of Extremely Antibiotic-Resistant Strains of Pseudomonas aeruginosa Isolated from Patients of a Clinic in Sincelejo, Colombia

by Nerlis Pajaro-Castro, Erick Diaz-Morales and Kenia Hoyos

BioTech 2025, 14(1), 21; https://doi.org/10.3390/biotech14010021 - 16 Mar 2025

Abstract

Pseudomonas aeruginosa is an opportunistic pathogen classified as a priority and a great public health concern; therefore, this research focuses on the genomic characterization of extremely resistant strains of P. aeruginosa isolated from patients in a clinic in Sincelejo, Colombia. Seven strains were [...] Read more.

Pseudomonas aeruginosa is an opportunistic pathogen classified as a priority and a great public health concern; therefore, this research focuses on the genomic characterization of extremely resistant strains of P. aeruginosa isolated from patients in a clinic in Sincelejo, Colombia. Seven strains were analyzed by whole genome sequencing using the Illumina NovaSeq platform, with a focus on the identification of resistance genes and virulence factors through the CARD and VFDB databases. An ANI (Average Nucleotide Identity) analysis was carried out to determine the genetic relationship between the strains, complemented by a phylogenomic analysis to place the strains in different evolutionary clades. The results revealed that six of the strains are of Colombian origin, while one strain (547256) belongs to the high-risk clone ST773, previously unidentified in Colombia. Genome size ranged from 6 to 7.4 Mbp, indicating differences in genetic content among strains. Phylogenomic analysis confirmed that five strains belong to a multidrug-resistant (MDR) group, while one strain (572897) showed high alignment with a laboratory strain, and strain 547256 was not associated with any specific clade. Clinically, 100% of strains showed carbapenem resistance, resistance genes, and virulence factors that make them difficult to treat. This study provides key insights into the genetic diversity and resistance patterns of P. aeruginosa in this region, underscoring the need to monitor high-risk clones and optimize therapeutic strategies. Full article

(This article belongs to the Section Computational Biology)

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13 pages, 3304 KiB

Open AccessArticle

Using Nano-Luciferase Binary (NanoBiT) Technology to Assess the Interaction Between Viral Spike Protein and Angiotensin-Converting Enzyme II by Aptamers

by Meng-Wei Lin, Cheng-Han Lin, Hua-Hsin Chiang, Irwin A. Quintela, Vivian C. H. Wu and Chih-Sheng Lin

BioTech 2025, 14(1), 20; https://doi.org/10.3390/biotech14010020 - 15 Mar 2025

Abstract

Nano-luciferase binary technology (NanoBiT)-based pseudoviral sensors are innovative tools for monitoring viral infection dynamics. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infects host cells via its trimeric surface spike protein, which binds to the human angiotensin-converting enzyme II (hACE2) receptor. This interaction is [...] Read more.

Nano-luciferase binary technology (NanoBiT)-based pseudoviral sensors are innovative tools for monitoring viral infection dynamics. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infects host cells via its trimeric surface spike protein, which binds to the human angiotensin-converting enzyme II (hACE2) receptor. This interaction is crucial for viral entry and serves as a key target for therapeutic interventions against coronavirus disease 2019 (COVID-19). Aptamers, short single-stranded DNA (ssDNA) or RNA molecules, are highly specific, high-affinity biorecognition elements for detecting infective pathogens. Despite their potential, optimizing viral infection assays using traditional protein–protein interaction (PPI) methods often face challenges in optimizing viral infection assays. In this study, we selected and evaluated aptamers for their ability to interact with viral proteins, enabling the dynamic visualization of infection progression. The NanoBiT-based pseudoviral sensor demonstrated a rapid increase in luminescence within 3 h, offering a real-time measure of viral infection. A comparison of detection technologies, including green fluorescent protein (GFP), luciferase, and NanoBiT technologies for detecting PPI between the pseudoviral spike protein and hACE2, highlighted NanoBiT’s superior sensitivity and performance, particularly in aptamer selection. This bioluminescent system provides a robust, sensitive, and early-stage quantitative approach to studying viral infection dynamics. Full article

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17 pages, 608 KiB

Open AccessArticle

A Coupled Model of the Cardiovascular and Immune Systems to Analyze the Effects of COVID-19 Infection

by Camila Ribeiro Passos, Alexandre Altamir Moreira, Ruy Freitas Reis, Rodrigo Weber dos Santos, Marcelo Lobosco and Bernardo Martins Rocha

BioTech 2025, 14(1), 19; https://doi.org/10.3390/biotech14010019 - 12 Mar 2025

Abstract

The COVID-19 pandemic has underscored the importance of understanding the interplay between the cardiovascular and immune systems during viral infections. SARS-CoV-2 enters human cells via the ACE-2 enzyme, initiating a cascade of immune responses. This study presents a coupled mathematical model that integrates [...] Read more.

The COVID-19 pandemic has underscored the importance of understanding the interplay between the cardiovascular and immune systems during viral infections. SARS-CoV-2 enters human cells via the ACE-2 enzyme, initiating a cascade of immune responses. This study presents a coupled mathematical model that integrates the cardiovascular system (CVS) and immune system (IS), capturing their complex interactions during infection. The CVS model, based on ordinary differential equations, describes heart dynamics and pulmonary and systemic circulation, while the IS model simulates immune responses to SARS-CoV-2, including immune cell interactions and cytokine production. A coupling strategy transfers information from the IS to the CVS at specific intervals, enabling the exploration of immune-driven cardiovascular effects. Numerical simulations examined how these interactions influence infection severity and recovery. The coupled model accurately replicated the evolution of cardiac function in survivors and non-survivors of COVID-19. Survivors exhibited a left ventricular ejection fraction (LVEF) reduction of up to

25 %

while remaining within normal limits, whereas non-survivors showed a severe 4-fold decline, indicative of myocardial dysfunction. Similarly, the right ventricular ejection fraction (RV EF) decreased by approximately

50 %

in survivors but underwent a drastic 5-fold reduction in non-survivors. These findings highlight the model’s capacity to distinguish differential cardiac dysfunction across clinical outcomes and its potential to enhance our understanding of COVID-19 pathophysiology. Full article

(This article belongs to the Section Computational Biology)

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14 pages, 3343 KiB

Open AccessArticle

Characterization of a GH43 Bifunctional Glycosidase from Endophytic Chaetomium globosum and Its Potential Application in the Biotransformation of Ginsenosides

by Yao Lu, Qiang Jiang, Yamin Dong, Runzhen Ji, Yiwen Xiao, Du Zhu and Boliang Gao

BioTech 2025, 14(1), 18; https://doi.org/10.3390/biotech14010018 - 12 Mar 2025

Abstract

The GH43 family of glycosidases represents an important class of industrial enzymes that are widely utilized across the food, pharmaceutical, and various other sectors. In this study, we identified a GH43 family glycoside hydrolytic enzyme, Xyaf313, derived from the plant endophytic fungus [...] Read more.

The GH43 family of glycosidases represents an important class of industrial enzymes that are widely utilized across the food, pharmaceutical, and various other sectors. In this study, we identified a GH43 family glycoside hydrolytic enzyme, Xyaf313, derived from the plant endophytic fungus Chaetomium globosum DX-THS3, which is capable of transforming several common ginsenosides. The enzyme function analysis reveals that Xyaf313 exhibits dual functionality, displaying both α-L-arabinofuranosidase and β-D-xylosidase activity. When acting as an α-L-arabinofuranosidase, Xyaf313 achieves optimal enzyme activity of 23.96 U/mg at a temperature of 50 °C and a pH of 7. In contrast, its β-D-xylosidase activity results in a slight reduction in enzyme activity to 23.24 U/mg, with similar optimal temperature and pH conditions to those observed for the α-L-arabinofuranosidase activity. Furthermore, Xyaf313 demonstrates considerable resistance to most metal ions and common chemical reagents. Notably, while the maximum enzyme activity of Xyaf313 occurs at 50 °C, it maintains high activity at room temperature (30 °C), with relative enzyme activity exceeding 90%. Measurements of ginsenoside transformation show that Xyaf313 can convert common ginsenosides Rc, Rb₁, Rb₂, and Rb₃ into Rd, underscoring its potential for pharmaceutical applications. Overall, our findings contribute to the identification of a new class of bifunctional GH43 glycoside hydrolases, highlight the significance of plant endophytic fungi as a promising resource for the screening of carbohydrate-decomposing enzymes, and present new candidate enzymes for the biotransformation of ginsenosides. Full article

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