BioTech

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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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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BioTech, Volume 14, Issue 2 (June 2025) – 12 articles

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