BioTech

15 pages, 2540 KiB

Open AccessArticle

Optimized Extraction of Polyphenols from Kiwifruit Peels and Their Biological Activities

by Batoul Shkeir, Nada El Darra, Bilal Azakir, Salma Khazaal, Elie Salem Sokhn, Mohamed Koubaa, Richard G. Maroun, Nicolas Louka and Espérance Debs

BioTech 2024, 13(4), 54; https://doi.org/10.3390/biotech13040054 - 2 Dec 2024

Abstract

(1) Background: Kiwifruit is a globally valued fruit. Its industrial processing produces a substantial amount of waste, particularly peels, which present an appealing potential source of bioactive compounds. This study focuses on optimizing the extraction of phenolics from kiwi peels using a water [...] Read more.

(1) Background: Kiwifruit is a globally valued fruit. Its industrial processing produces a substantial amount of waste, particularly peels, which present an appealing potential source of bioactive compounds. This study focuses on optimizing the extraction of phenolics from kiwi peels using a water bath (WB) and infrared irradiation (IR) and assessing their biological activities. (2) Methods: Optimal conditions for polyphenol extraction from kiwifruit peels, in terms of temperature and time, were determined using Response Surface Methodology. Total phenolic content (TPC) was measured by the Folin–Ciocalteu method, and antioxidant activity was assessed utilizing the DPPH method. Antibacterial activities against Bacillus cereus, Staphylococcus aureus, Escherichia coli, and Salmonella Typhimurium were determined using the minimum inhibitory concentration (MIC). The lyophilized extract was tested for its anticancer effect on A549 lung cancer cell lines. The phytochemical profiles of the WB and IR extracts were analyzed through High-Performance Liquid Chromatography (HPLC). (3) Results: The optimal extraction conditions for the WB method were found to be 75 °C for 35 min, and 90 °C for 5 min for IR. The corresponding TPC obtained by IR was 21 mg GAE/g DM, which was 4.4 times higher than that obtained by WB (4.8 mg GAE/g DM). This indicates that IR was more efficient in extracting phenolics from kiwi peels. The antioxidant activity under the optimum conditions of WB and IR was 14 mg TE/g DM and 16 mg TE/g DM, respectively. Both the WB and IR extracts demonstrated antibacterial activity against B. cereus with an MIC value of 25 mg/mL. Additionally, the IR extract displayed an antibacterial effect against S. aureus, with an MIC value of 50 mg/mL. The WB and IR kiwi peel extracts were effective in significantly reducing A549 cell viability at 50 µg/mL and 100 µg/mL, respectively. Caffeic acid (0.54 ppm) and catechin (5.44 ppm) were the major polyphenols identified in WB and IR extracts, as determined by HPLC. (4) Conclusions: IR was more efficient in extracting phenolics from kiwi peels than WB. The findings also suggest that kiwi peel can be effectively utilized as an antioxidant, antibacterial, and anticancer agent. Full article

(This article belongs to the Section Agricultural and Food Biotechnology)

► Show Figures

Figure 1

19 pages, 3417 KiB

Open AccessReview

Prospects in the Use of Cannabis sativa Extracts in Nanoemulsions

by Ian Vitola, Carlos Angulo, Raul C. Baptista-Rosas, Luis Miguel Anaya-Esparza, Zazil Yadel Escalante-García, Angélica Villarruel-López and Jorge Manuel Silva-Jara

BioTech 2024, 13(4), 53; https://doi.org/10.3390/biotech13040053 - 2 Dec 2024

Abstract

Cannabis sativa plants have been widely investigated for their specific compounds with medicinal properties. These bioactive compounds exert preventive and curative effects on non-communicable and infectious diseases. However, C. sativa extracts have barely been investigated, although they constitute an affordable option to treat [...] Read more.

Cannabis sativa plants have been widely investigated for their specific compounds with medicinal properties. These bioactive compounds exert preventive and curative effects on non-communicable and infectious diseases. However, C. sativa extracts have barely been investigated, although they constitute an affordable option to treat human diseases. Nonetheless, antioxidant, antimicrobial, and immunogenicity effects have been associated with C. sativa extracts. Furthermore, innovative extraction methods in combination with nanoformulations have been proposed to increase desirable compounds’ availability, distribution, and conservation, which can be aided by modern computational tools in a transdisciplinary approach. This review aims to describe available extraction and nanoformulation methods for C. sativa, as well as its known antioxidant, antimicrobial, and immunogenic activities. Critical points on the use of C. sativa extracts in nanoformulations are identified and some prospects are envisaged. Full article

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

► Show Figures

Figure 1

11 pages, 3213 KiB

Open AccessCommunication

Composition, Seasonal Dynamics and Metabolic Potential of the Rhizosphere Microbiome Associated with Wild White Poplar

by Mikhail I. Popchenko, Dmitry S. Karpov, Natalya S. Gladysh, Maxim A. Kovalev, Vsevolod V. Volodin, George S. Krasnov, Alina S. Bogdanova, Nadezhda L. Bolsheva, Maria S. Fedorova and Anna V. Kudryavtseva

BioTech 2024, 13(4), 52; https://doi.org/10.3390/biotech13040052 - 1 Dec 2024

Abstract

The white poplar (Populus alba) is a dioecious woody plant with significant potential for the phytoremediation of soils. To realize this potential, it is necessary to utilize growth-promoting microorganisms. One potential source of such beneficial microorganisms is the rhizosphere community of [...] Read more.

The white poplar (Populus alba) is a dioecious woody plant with significant potential for the phytoremediation of soils. To realize this potential, it is necessary to utilize growth-promoting microorganisms. One potential source of such beneficial microorganisms is the rhizosphere community of wild-growing trees. However, the structure, dynamics, and metabolism of the rhizosphere community of wild-growing white poplar remain poorly understood. To ascertain seasonal dynamics, species diversity, and metabolic potential, we sequenced 16S rRNA genes in metagenomes derived from 165 soil samples collected in spring and autumn from the root surfaces of 102 trees situated in disparate geographical locations. The three most prevalent phyla across all samples are Proteobacteria, Actinobacteriota, and Acidobacteriota. At the order level, the most prevalent orders are Sphingomonadales and Rhizobiales. Accordingly, the families Sphingomonadaceae and Rhizobiaceae were identified as dominant. The rhizospheric microbiome exhibited substantial inter-seasonal variation. Six families, including Caulobacteraceae, Xanthomonadaceae, Chitinophagaceae, Chthoniobacteraceae, Sphingomonadaceae, and Rhizobiaceae, exhibited alterations (spring-to-autumn) across all geographical locations under study. Members of the Rhizobiaceae family, which includes nitrogen-fixing bacteria, can provide poplar with plant-available forms of nitrogen such as nitrate and ammonium. The rhizosphere microbiome may facilitate the conversion of inorganic sulfur into sulfur-containing amino acids, cysteine and methionine, that are bioavailable to plants. Furthermore, the rhizosphere microbiome is capable of synthesizing amino acids, organic acids (including Krebs cycle acids), and some lipids and sugars. Consequently, the rhizosphere community can stimulate poplar growth by providing it with readily available forms of nitrogen and sulfur, as well as building blocks for the synthesis of proteins, nucleic acids, and other macromolecules. Many of these pathways, including nitrogen fixation, were subjected to seasonal changes. Full article

► Show Figures

Figure 1

12 pages, 1608 KiB

Open AccessReview

The Biotechnological Potential of Crickets as a Sustainable Protein Source for Fishmeal Replacement in Aquafeed

by Aldo Fraijo-Valenzuela, Joe Luis Arias-Moscoso, Oscar Daniel García-Pérez, Libia Zulema Rodriguez-Anaya and Jose Reyes Gonzalez-Galaviz

BioTech 2024, 13(4), 51; https://doi.org/10.3390/biotech13040051 - 21 Nov 2024

Abstract

As aquaculture production grows, so does the demand for quality and cost-effective protein sources. The cost of fishmeal (FM) has increased over the years, leading to increased production costs for formulated aquafeed. Soybean meal (SBM) is commonly used as an FM replacer in [...] Read more.

As aquaculture production grows, so does the demand for quality and cost-effective protein sources. The cost of fishmeal (FM) has increased over the years, leading to increased production costs for formulated aquafeed. Soybean meal (SBM) is commonly used as an FM replacer in aquafeed, but anti-nutritional factors could affect the growth, nutrition, and health of aquatic organisms. Cricket meal (CM) is an alternative source with a nutrient profile comparable to FM due to its high protein content, digestibility, and amino acid profile. CM use in aquafeed influences growth and reproductive performance while modulating the gut microbiota and immune response of fish and shrimp. However, consistent regulation and scaling up are necessary for competitive prices and the marketing of CM. Moreover, the chitin content in CM could be an issue in some fish species; however, different strategies based on food biotechnology can improve the protein quality for its safe use in aquafeed. Full article

(This article belongs to the Section Agricultural and Food Biotechnology)

► Show Figures

Figure 1

14 pages, 2500 KiB

Open AccessArticle

Leveraging Walnut Somatic Embryos as a Biomanufacturing Platform for Recombinant Proteins and Metabolites

by Paulo A. Zaini, Katherine R. Haddad, Noah G. Feinberg, Yakir Ophir, Somen Nandi, Karen A. McDonald and Abhaya M. Dandekar

BioTech 2024, 13(4), 50; https://doi.org/10.3390/biotech13040050 - 15 Nov 2024

Abstract

Biomanufacturing enables novel sources of compounds with constant demand, such as food coloring and preservatives, as well as new compounds with peak demand, such as diagnostics and vaccines. The COVID-19 pandemic has highlighted the need for alternative sources of research materials, thrusting research [...] Read more.

Biomanufacturing enables novel sources of compounds with constant demand, such as food coloring and preservatives, as well as new compounds with peak demand, such as diagnostics and vaccines. The COVID-19 pandemic has highlighted the need for alternative sources of research materials, thrusting research on diversification of biomanufacturing platforms. Here, we show initial results exploring the walnut somatic embryogenic system expressing the recombinant receptor binding domain (RBD) and ectodomain of the spike protein (Spike) from the SARS-CoV-2 virus. Stably transformed walnut embryo lines were selected and propagated in vitro. Both recombinant proteins were detected at 3–14 µg/g dry weight of tissue culture material. Although higher yields of recombinant protein have been obtained using more conventional biomanufacturing platforms, we also report on the production of the red pigment betanin in somatic embryos, reaching yields of 650 mg/g, even higher than red beet Beta vulgaris. This first iteration shows the potential of biomanufacturing using somatic walnut embryos that can now be further optimized for different applications sourcing specialized proteins and metabolites. Full article

► Show Figures

Figure 1

18 pages, 518 KiB

Open AccessReview

Climate Resilience in Farm Animals: Transcriptomics-Based Alterations in Differentially Expressed Genes and Stress Pathways

by Chikamagalore Gopalakrishna Shashank, Veerasamy Sejian, Mullakkalparambil Velayudhan Silpa, Chinnasamy Devaraj, Aradotlu Parameshwarappa Madhusoodan, Ebenezer Binuni Rebez, Gajendirane Kalaignazhal, Artabandhu Sahoo and Frank Rowland Dunshea

BioTech 2024, 13(4), 49; https://doi.org/10.3390/biotech13040049 - 15 Nov 2024

Abstract

The livestock sector, essential for maintaining food supply and security, encounters numerous obstacles as a result of climate change. Rising global populations exacerbate competition for natural resources, affecting feed quality and availability, heightening livestock disease risks, increasing heat stress, and contributing to biodiversity [...] Read more.

The livestock sector, essential for maintaining food supply and security, encounters numerous obstacles as a result of climate change. Rising global populations exacerbate competition for natural resources, affecting feed quality and availability, heightening livestock disease risks, increasing heat stress, and contributing to biodiversity loss. Although various management and dietary interventions exist to alleviate these impacts, they often offer only short-lived solutions. We must take a more comprehensive approach to understanding how animals adapt to and endure their environments. One such approach is quantifying transcriptomes under different environments, which can uncover underlying pathways essential for livestock adaptation. This review explores the progress and techniques in studies that apply gene expression analysis to livestock production systems, focusing on their adaptation to climate change. We also attempt to identify various biomarkers and transcriptomic differences between species and pure/crossbred animals. Looking ahead, integrating emerging technologies such as spatialomics could further accelerate genetic improvements, enabling more thermoresilient and productive livestock in response to future climate fluctuations. Ultimately, insights from these studies will help optimize livestock production systems by identifying thermoresilient/desired animals for use in precise breeding programs to counter climate change. Full article

(This article belongs to the Section Agricultural and Food Biotechnology)

► Show Figures

Figure 1

16 pages, 2975 KiB

Open AccessArticle

High-Resolution Melting Analysis Potential for Saccharomyces cerevisiae var. boulardii Authentication in Probiotic-Enriched Food Matrices

by Monika Borkowska, Michał Kułakowski and Kamila Myszka

BioTech 2024, 13(4), 48; https://doi.org/10.3390/biotech13040048 - 14 Nov 2024

Abstract

To date, the only probiotic yeast with evidence of health-promoting effects is Saccharomyces cerevisiae var. boulardii. The expanded market including dietary supplements and functional foods supplemented with Saccharomyces cerevisiae var. boulardii creates an environment conductive to food adulterations, necessitating rapid testing to verify [...] Read more.

To date, the only probiotic yeast with evidence of health-promoting effects is Saccharomyces cerevisiae var. boulardii. The expanded market including dietary supplements and functional foods supplemented with Saccharomyces cerevisiae var. boulardii creates an environment conductive to food adulterations, necessitating rapid testing to verify product probiotic status. Herein, qPCR-HRM analysis was tested for probiotic yeast identification. The effectiveness of the primer pairs’ set was examined, designed to amplify heterogeneous regions in (a) rDNA sequences previously designed to identify food-derived yeast and (b) genes associated with physiological and genotypic divergence of Saccharomyces cerevisiae var. boulardii. Preliminary tests of amplicons’ differentiation power enabled the selection of interspecies sequences for 18SrRNA and ITS and genus-specific sequences HO, RPB2, HXT9 and MAL11. The multi-fragment qPCR-HRM analysis was sufficient for culture-dependent Saccharomyces cerevisiae var. boulardii identification and proved effective in the authentication of dietary supplements’ probiotic composition. The identification of S. cerevisiae var. boulardii in complex microbial mixtures of kefir succeeded with more specific intragenus sequences HO and RPB2. The predominance of S. cerevisiae var. boulardii in the tested matrices, quantitatively corresponded to the probiotic-enriched food, was crucial for identification with qPCR–HRM analysis. Considering the reported assumptions, qPCR-HRM analysis is an appropriate tool for verifying probiotic-enriched food. Full article

(This article belongs to the Section Agricultural and Food Biotechnology)

► Show Figures

Figure 1

18 pages, 3391 KiB

Open AccessReview

Honey Bioactive Molecules: There Is a World Beyond the Sugars

by Gregorio Bonsignore, Simona Martinotti and Elia Ranzato

BioTech 2024, 13(4), 47; https://doi.org/10.3390/biotech13040047 - 14 Nov 2024

Abstract

Honey’s many bioactive compounds have been utilized historically to cure infectious diseases. Beneficial effects are its antiviral, antibacterial, anti-inflammatory, antioxidant, and immune-stimulating qualities. The bee species, geographic location, botanical origin, harvest season, processing, and storage conditions all affect honey’s potential for therapeutic use. [...] Read more.

Honey’s many bioactive compounds have been utilized historically to cure infectious diseases. Beneficial effects are its antiviral, antibacterial, anti-inflammatory, antioxidant, and immune-stimulating qualities. The bee species, geographic location, botanical origin, harvest season, processing, and storage conditions all affect honey’s potential for therapeutic use. Honey contains a number of antioxidants and active compounds, such as polyphenols, which have been shown to have disease-preventive properties. Based on their origins, categories, and functions, the main polyphenols found in various honey varieties are examined in this review. Full article

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

► Show Figures

Figure 1

17 pages, 5204 KiB

Open AccessArticle

Screening and Characterization of Sialic Acid-Binding Variable Lymphocyte Receptors from Hagfish

by Mark Rickard N. Angelia, Abigail Joy D. Rodelas-Angelia, Cheolung Yang, Sojeong Park, Seung pyo Jeong, Hyeok Jang, Dennis Berbulla Bela-ong, Hobin Jang, Kim D. Thompson and Taesung Jung

BioTech 2024, 13(4), 46; https://doi.org/10.3390/biotech13040046 - 12 Nov 2024

Abstract

Sialic acid is a diverse group of monosaccharides often found on the termini of N- and O-linked glycans as well as being components of glycoconjugates. Hypersialylation has been associated with the progression of chronic inflammation-mediated diseases such as cardiovascular disease and [...] Read more.

Sialic acid is a diverse group of monosaccharides often found on the termini of N- and O-linked glycans as well as being components of glycoconjugates. Hypersialylation has been associated with the progression of chronic inflammation-mediated diseases such as cardiovascular disease and cancer. Given its role in infection and disease-related processes, sialic acid is a promising target for therapeutic approaches that utilize carbohydrate-binding molecules. In this study, we screened for sialic acid-recognizing variable lymphocyte receptors (VLRBs) or ccombodies from inshore hagfish (Eptatretus burgeri) using a synthetic Neu5Ac-glycoconjugate as an antigen in immunoassay. Resulting ccombodies, 2D8, 5G11, 4A1, and 5F8 were further characterized in terms of their binding activity and specificity. A competitive ELISA using free haptens showed strong inhibition using either N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc). The half-maximal inhibitory concentrations (IC₅₀) for Neu5Ac ranged from 7.02 to 17.06 mM, with candidates 4A1 and 5G11 requiring the least and highest amounts, respectively. IC₅₀ values for Neu5Gc ranged from 8.12 to 13.91 mM, for 4A1 and 5G11, respectively. Candidate ccombodies also detected naturally occurring sialic acid from known sialoglycoproteins using a dot blot assay. Neu5Gc-5G11 and Neu5Ac-2D8 yielded the strongest and weakest docking interactions with affinity values of −5.9 kcal/mol and −4.9 kcal/mol, respectively. Hydrogen bonding and hydrophobic interactions were predicted to be the predominant noncovalent forces observed between the ccombodies and sialic acid. This study demonstrates that glycan-binding VLRBs from hagfish hold promise in augmenting the glycobiologists’ toolkit in investigating the roles of glycans in human and animal health and disease. Full article

(This article belongs to the Section Agricultural and Food Biotechnology)

► Show Figures

Figure 1

16 pages, 2544 KiB

Open AccessReview

Artificial Insemination as a Possible Convenient Tool to Acquire Genome-Edited Mice via In Vivo Fertilization with Engineered Sperm

by Masahiro Sato, Emi Inada, Issei Saitoh, Kazunori Morohoshi and Shingo Nakamura

BioTech 2024, 13(4), 45; https://doi.org/10.3390/biotech13040045 - 11 Nov 2024

Abstract

Advances in genome editing technology have made it possible to create genome-edited (GE) animals, which are useful for identifying isolated genes and producing models of human diseases within a short period of time. The production of GE animals mainly relies on the gene [...] Read more.

Advances in genome editing technology have made it possible to create genome-edited (GE) animals, which are useful for identifying isolated genes and producing models of human diseases within a short period of time. The production of GE animals mainly relies on the gene manipulation of pre-implantation embryos, such as fertilized eggs and two-cell embryos, which can usually be achieved by the microinjection of nucleic acids, electroporation in the presence of nucleic acids, or infection with viral vectors, such as adeno-associated viruses. In contrast, GE animals can theoretically be generated by fertilizing ovulated oocytes with GE sperm. However, there are only a few reports showing the successful production of GE animals using GE sperm. Artificial insemination (AI) is an assisted reproduction technology based on the introduction of isolated sperm into the female reproductive tract, such as the uterine horn or oviductal lumen, for the in vivo fertilization of ovulated oocytes. This approach is simpler than the in vitro fertilization-based production of offspring, as the latter always requires an egg transfer to recipient females, which is labor-intensive and time-consuming. In this review, we summarize the various methods for AI reported so far, the history of sperm-mediated gene transfer, a technology to produce genetically engineered animals through in vivo fertilization with sperm carrying exogenous DNA, and finally describe the possibility of AI-mediated creation of GE animals using GE sperm. Full article

(This article belongs to the Section Biotechnology Regulation)

► Show Figures

Graphical abstract

16 pages, 3361 KiB

Open AccessArticle

Ultraviolet, Did the Cell See It from the Side or the Bottom? Assessment and Modeling of UV Effects on Cultured Cells Using the CL-1000 UV-Crosslinker

by Takahiro Oyama, Kai Yanagihara, Anna Arai, Takanori Kamiya, Midori Oyama, Takashi Tanikawa, Takehiko Abe and Tomomi Hatanaka

BioTech 2024, 13(4), 44; https://doi.org/10.3390/biotech13040044 - 25 Oct 2024

Abstract

Numerous natural extracts and compounds have been evaluated for their ability to mitigate the adverse effects of ultraviolet (UV) overexposure. However, variability in the UV doses that trigger biological responses across studies likely arises from inconsistencies in UV exposure standardization. We hypothesize that [...] Read more.

Numerous natural extracts and compounds have been evaluated for their ability to mitigate the adverse effects of ultraviolet (UV) overexposure. However, variability in the UV doses that trigger biological responses across studies likely arises from inconsistencies in UV exposure standardization. We hypothesize that these discrepancies are due to variations in culture plates and dishes. The UV dose (D) required to reduce cell viability by 50% differed by a factor of ten between 3.5 cm dishes and 96-well plates. Similarly, the EC₅₀ dose for IL-6 release (D_1/2) varied, potentially correlating with the surface area (S). UV exposure to wells with increasing height in 3.5 cm dishes resulted in a decrease in IL-6 release, suggesting that the greater the well height, the more it may influence UV exposure through reflection or shielding effects, thereby contributing to the physiological effects on the cells. To compare these differences among plates, we defined the height-to-diameter ratio (r). Analysis revealed a linear correlation between D_1/2 and S in a log-log plot, and between D_1/2 and r in a semi-log plot. From this, we defined two empirical indices σ and ρ for UV dose adjustment. A deductive model was also developed to derive a D′ value that adjusts UV doses without requiring training. As with σ and ρ, the UV dose D was effectively adjusted using D′ as well. These attempts suggest that D′ offers a foundational framework for evaluating UVB effects on cultured cells. Full article

► Show Figures

Figure 1

13 pages, 1793 KiB

Open AccessArticle

Microbial Protein and Metabolite Profiles of Klebsiella oxytoca M5A1 in a Bubble Column Bioreactor

by Tawakalt Ayodele, Musiliu Liadi, Abodunrin Tirmidhi Tijani, Kudirat Alarape, Christiana Bitrus, Clairmont L. Clementson and Ademola Hammed

BioTech 2024, 13(4), 43; https://doi.org/10.3390/biotech13040043 - 19 Oct 2024

Abstract

The production of microbial proteins (MPs) has emerged as a critical focus in biotechnology, driven by the need for sustainable and scalable alternatives to traditional protein sources. This study investigates the efficacy of two experimental setups in producing MPs using the nitrogen-fixing bacterium [...] Read more.

The production of microbial proteins (MPs) has emerged as a critical focus in biotechnology, driven by the need for sustainable and scalable alternatives to traditional protein sources. This study investigates the efficacy of two experimental setups in producing MPs using the nitrogen-fixing bacterium Klebsiella oxytoca M5A1. K. oxytoca M5A1, known for its facultative anaerobic growth and capability to fix atmospheric nitrogen, offers a promising avenue for environmentally friendly protein production. This research compares the performance of a simple bubble column (BC) bioreactor, which promotes efficient mixing and cross-membrane gas transfer, with static fermentation, a traditional method lacking agitation and aeration. The study involved the parallel cultivation of K. oxytoca M5A1 in both systems, with key parameters such as microbial growth, glucose utilization, protein concentration, and metabolite profiles monitored over a 48 h period. The results indicate that the BC bioreactor consistently outperformed static fermentation regarding the growth rate, protein yield, and glucose utilization efficiency. The BC exhibited a significant increase in protein production, reaching 299.90 µg/mL at 48 h, compared to 219.44 µg/mL in static fermentation. The organic acid profile reveals both synthesis and utilization regimes of varying patterns. These findings highlight the advantages of the BC bioreactor for MP production, particularly its ability to maintain aerobic conditions that support higher growth and yield. Full article

(This article belongs to the Section Industrial Biotechnology)

► Show Figures

Figure 1

18 pages, 3640 KiB

Open AccessArticle

Phytochemical Composition, Antioxidant, Anti-Helicobacter pylori, and Enzyme Inhibitory Evaluations of Cleistocalyx operculatus Flower Bud and Leaf Fractions

by Doan Thien Thanh, Mai Thanh Tan, Nguyen Thi My Thu, Pham Nhat Phuong Trinh, Pham Thi Hoai Thuong, Pham Thi Giang Tuyet, Luong Thi My Ngan and Tran Trung Hieu

BioTech 2024, 13(4), 42; https://doi.org/10.3390/biotech13040042 - 11 Oct 2024

Abstract

Six solvent fractions isolated from flower bud and leaf ethanolic extracts of Cleistocalyx operculatus were analyzed for their phytochemical contents, including phenolics, flavonoids, saponins, tannins, and alkaloids. Antioxidant activities were measured using the ABTS, DPPH, and FRAP assays. The results showed that the [...] Read more.

Six solvent fractions isolated from flower bud and leaf ethanolic extracts of Cleistocalyx operculatus were analyzed for their phytochemical contents, including phenolics, flavonoids, saponins, tannins, and alkaloids. Antioxidant activities were measured using the ABTS, DPPH, and FRAP assays. The results showed that the flower bud aqueous fraction (BAF) and the leaf aqueous fraction (LAF) rich in phenolic content (768.18 and 490.74 mg GAE/g dry extract, respectively) exhibited significantly higher antioxidant activities than the other fractions. The flower bud hexane fraction (BHF) had remarkably high flavonoid and saponin contents (134.77 mg QE/g and 153.33 mg OA/g dry extract, respectively), followed by that of the leaf hexane fraction (LHF) (76.54 mg QE/g and 88.25 mg OA/g dry extract, respectively). The BHF and LHF were found to have extremely high antibacterial activity against two H. pylori strains, ATCC 51932 and 43504 (MICs of 125 µg/mL). Interestingly, DMC (2′,4′-Dihydroxy-6′-methoxy-3′,5′-dimethylchalcone) isolated from the BHF displayed greater antibacterial activity against the bacterial strains (MICs of 25–50 µg/mL) than those of the fractions. In addition, DMC presented potent inhibitory effects on H. pylori urease (IC₅₀ of 3.2 µg/mL) and α-amylase (IC₅₀ of 83.80 µg/mL), but no inhibition against α-glucosidase. It was also demonstrated that DMC showed pronounced inhibitory effects on the urease activity and biofilm formation of H. pylori, and could increase the membrane permeability of the bacterial cells. Scanning electron micrographs depicted that the BHF and DMC had strong effects on the cell shape and significantly induced the distortion and damage of the cell membrane. The fractions and DMC showed no significant toxicity to four tested human cell lines. Efforts to reduce antibiotic use indicate the need for further studies of the flower buds and DMC as potential products to prevent or treat gastric H. pylori infections. Full article

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

► Show Figures

Graphical abstract

16 pages, 2581 KiB

Open AccessArticle

Loss of Cell-Cell Contact Inhibits Cellular Differentiation of α-Catenin Knock Out P19 Embryonal Carcinoma Cells and Their Colonization into the Developing Mouse Embryos

by Masahiro Sato, Emi Inada, Naoko Kubota and Masayuki Ozawa

BioTech 2024, 13(4), 41; https://doi.org/10.3390/biotech13040041 - 3 Oct 2024

Abstract

Cadherin−catenin cell−cell adhesion complexes, composed of cadherin, β-catenin or plakoglobin, and α-catenin (α-cat) molecules, are crucial for maintaining cell−cell contact and are commonly referred to as “adherens junctions (AJs).” Inactivating this system leads to loss of cell−cell contact and developmental arrest in early [...] Read more.

Cadherin−catenin cell−cell adhesion complexes, composed of cadherin, β-catenin or plakoglobin, and α-catenin (α-cat) molecules, are crucial for maintaining cell−cell contact and are commonly referred to as “adherens junctions (AJs).” Inactivating this system leads to loss of cell−cell contact and developmental arrest in early embryos. However, it remains unclear whether the loss of cell−cell contact affects the differentiation of embryonic cells. In this study, we explored the use of a murine embryonal carcinoma cell line, P19, as an in vitro model for early embryogenesis. P19 cells easily form embryoid bodies (EBs) and are susceptible to cellular differentiation in response to retinoic acid (RA) and teratoma formation. Using CRISPR/Cas9 technology to disrupt the endogenous α-cat gene in P19 cells, we generated α-cat knockout (KO) cells that exhibited a loss of cell−cell contact. When cultivated on non-coated dishes, these α-cat KO cells formed EBs, but their structures were labile. In the RA-containing medium, the α-cat KO EBs failed to produce differentiated cells on their outer layer and continued to express SSEA-1, an antigen specific to pluripotent cells. Teratoma formation assays revealed an absence of overt differentiated cells in tumors derived from α-cat KO P19 cells. Aggregation assays revealed the inability of the KO cells to colonize into the zona pellucida-denuded 8-cell embryos. These findings suggest that the AJs are essential for promoting the early stages of cellular differentiation and for the colonization of early-developing embryos. Full article

► Show Figures

Graphical abstract

16 pages, 2552 KiB

Open AccessArticle

Structural Analysis and Substrate Specificity of D-Carbamoylase from Pseudomonas

by Marina Paronyan, Haykanush Koloyan, Hovsep Aganyants, Artur Hambardzumyan, Tigran Soghomonyan, Sona Avetisyan, Sergey Kocharov, Henry Panosyan, Vehary Sakanyan and Anichka Hovsepyan

BioTech 2024, 13(4), 40; https://doi.org/10.3390/biotech13040040 - 3 Oct 2024

Abstract

The synthesis of enantiomeric forms of D-amino acids can be achieved by a two-step “hydantoinase process” based on the sequential catalysis of substrates by specific enzymes, D-carbamoylase and D-hydantoinase. Here, we describe the structural features of D-carbamoylase from Pseudomonas, the encoded gene [...] Read more.

The synthesis of enantiomeric forms of D-amino acids can be achieved by a two-step “hydantoinase process” based on the sequential catalysis of substrates by specific enzymes, D-carbamoylase and D-hydantoinase. Here, we describe the structural features of D-carbamoylase from Pseudomonas, the encoded gene of which was chemically synthesized and cloned into Escherichia coli. A significant fraction of the overexpressed recombinant protein forms insoluble inclusion bodies, which are partially converted to a soluble state upon treatment with N-lauroylsarcosine or upon incubation of cells at 28 °C. Purified His-tagged protein exhibits the highest activity towards N-carbamoyl-D-alanine and N-carbamoyl-D-tryptophan. Comprehensive virtual analysis of the interactions of bulky carbamylated amino acids with D-carbamoylase provided valuable information. Molecular docking analysis revealed the location of the substrate binding site in the three-dimensional structure of D-carbamoylase. Molecular dynamics simulations showed that the binding pocket of the enzyme in complex with N-carbamoyl-D-tryptophan was stabilized within 100 nanoseconds. The free energy data showed that Arg176 and Asn173 formed hydrogen bonds between the enzyme and substrates. The studies of D-carbamoylases and the properties of our previously obtained D-hydantoinase suggest the possibility of developing a harmonized biotechnological process for the production of new drugs and peptide hormones. Full article

(This article belongs to the Topic Microbial Biotechnology Products and Biocatalysis Processes for a Sustainable Bioeconomy)

► Show Figures

Graphical abstract

19 pages, 34131 KiB

Open AccessArticle

Optimization of Sugar Extraction Process from Date Waste Using Full Factorial Design Toward Its Use for New Biotechnological Applications

by Islam Sayah, Mondher Njehi, Nicola Cicero, Vincenzo Nava, Manel Ben M’hadheb, Hatem Majdoub, Sami Achour and Teresa Gervasi

BioTech 2024, 13(4), 39; https://doi.org/10.3390/biotech13040039 - 3 Oct 2024

Abstract

In Tunisia, the date industry generates a large quantity of waste, raising environmental concerns. However, dates are rich in sugars, which offer a renewable source of nutrients for various applications. In this study, sugar extraction from two low-grade pitted date fruits (Alig and [...] Read more.

In Tunisia, the date industry generates a large quantity of waste, raising environmental concerns. However, dates are rich in sugars, which offer a renewable source of nutrients for various applications. In this study, sugar extraction from two low-grade pitted date fruits (Alig and Kentichi) under ultrasound, was optimized using full factorial design. At 40 °C, for20 min, and with a liquid-to-solid ratio of 10 mL/g, the optimum sugar contents were 60.87% and 50.79% for the varieties Alig and Kentichi, respectively. The date extracts were chemically analyzed, revealing low fat and protein contents, but significant polyphenol and mineral contents in both varieties. HPLC-IR analysis revealed more inverted sugars (glucose and fructose) in the Alig variety and more sucrose in the Kentichi variety. FTIR and SEM analysis showed the efficiency of the ultrasonic treatment of the biomass in terms of improving mass transfer diffusion through ultrasonic cavitation. Thus, ultrasound-assisted extraction constitutes an effective method for the recovery of sugar from date waste. Full article

(This article belongs to the Section Agricultural and Food Biotechnology)

► Show Figures

Figure 1

18 pages, 1912 KiB

Open AccessArticle

Chemical, Textural and Antioxidant Properties of Oat-Fermented Beverages with Different Starter Lactic Acid Bacteria and Pectin

by Dmitrii V. Khrundin and Elena V. Nikitina

BioTech 2024, 13(4), 38; https://doi.org/10.3390/biotech13040038 - 25 Sep 2024

Abstract

Currently, starter cultures for fermenting plant-based beverages are not widely available commercially, but producers can use starter cultures for dairy products. Therefore, the aim of this study was to determine the physicochemical, rheological, antioxidant and sensory properties of oat beverages with/without pectin fermented [...] Read more.

Currently, starter cultures for fermenting plant-based beverages are not widely available commercially, but producers can use starter cultures for dairy products. Therefore, the aim of this study was to determine the physicochemical, rheological, antioxidant and sensory properties of oat beverages with/without pectin fermented by four different dairy starter cultures. The use of a mono-starter with Lactobacillus bulgaricus or Sreptococcus thermophilus allows for the efficient use of glucose, and more lactic acid is accumulated. The beverage with L. bulgaricus is characterised by high adhesion, syneresis and low cohesiveness, and it has high antioxidant activity and a low sensory profile. Using starter with L. bulgaricus, S. thermophilus and some Lactococcus for fermentation yields a product with high sensory capacity, forming a high-viscosity beverage matrix with low syneresis, high water retention, chewy texture and stickiness. It has been observed that the absence of lactococci and the presence of Lactobacillus casei, L. Rhamnosus and L. paracasei in the starter yields a product with high antioxidant activity, especially in the presence of pectin. The use of pectin significantly improves the viscosity and textural properties of oat yoghurt, enhancing the drink’s flavour and giving it body. For many reasons, the use of different commercial starters in the dairy industry results in different viscosities of oat fermented beverages, forming a matrix with different textural, sensory and antioxidant properties. Full article

(This article belongs to the Section Agricultural and Food Biotechnology)

► Show Figures

Figure 1

18 pages, 3899 KiB

Open AccessArticle

The Elimination of Viroids through In Vitro Thermotherapy and a Meristem Tip Culture from a New Limonime Hybrid (Citrus x limon var. limon (L.) Burm. f. x Citrus latifolia var. latifolia)

by Virginia Sarropoulou, Katerina Grigoriadou, Varvara I. Maliogka, Chrysoula-Lito Sassalou and Vasileios Ziogas

BioTech 2024, 13(3), 37; https://doi.org/10.3390/biotech13030037 - 23 Sep 2024

Abstract

Viruses and viroids pose a significant challenge in citriculture, and their control is crucial for plant health. This study evaluated the effectiveness of in vitro thermotherapy combined with a meristem tip culture for eliminating citrus exocortis viroid (CEVd) and hop stunt viroid (HSVd) [...] Read more.

Viruses and viroids pose a significant challenge in citriculture, and their control is crucial for plant health. This study evaluated the effectiveness of in vitro thermotherapy combined with a meristem tip culture for eliminating citrus exocortis viroid (CEVd) and hop stunt viroid (HSVd) from a new limonime hybrid (Citrus x limon var. limon x Citrus latifolia var. latifolia). The elimination success was confirmed by RT-PCR assays. The in vitro elimination rate for CEVd during the shoot proliferation stage (43%) was higher than for HSVd (21%). Accordingly, in the subsequent rooting stage, the in vitro elimination rate for CEVd (50%) was higher than for HSVd (33%). Successful CEVd and HSVd eradication at a 100% rate was confirmed in the ex vitro acclimatized plants in the greenhouse. The study also established an efficient micropropagation protocol. The optimal treatment for in vitro shoot induction was 0.5–2 mg L⁻¹ benzyladenine (BA) + 0.5 mg L⁻¹ gibberellic acid (GA₃) + 0.25 mg L⁻¹ naphthalene acetic acid (NAA), while for shoot elongation, it was 0.5 mg L⁻¹ BA + 0.5 mg L⁻¹ kinetin (KIN) + 0.5 mg L⁻¹ GA₃ + 0.25 mg L⁻¹ NAA. Rooting was best promoted by 1 mg L⁻¹ NAA. This study provides valuable insights for the mass production of viroid-free propagation material in this new lemon x lime hybrid, contributing to the conservation of genetic resources in citrus breeding programs through the combined application of in vitro thermotherapy and an in vitro meristem tip culture, a novel and highlighted achievement reported for the first time in this study. Full article

(This article belongs to the Section Agricultural and Food Biotechnology)

► Show Figures

Figure 1

13 pages, 1397 KiB

Open AccessArticle

Clinical Evaluation of AMNIODERM+^® Wound Dressing Containing Non-Viable Human Amniotic Membrane: Retrospective-Perspective Clinical Trial

by Iveta Schmiedova, Petr Slama, Alena Dembickaja, Beata Kozova, Vendula Hyneckova, Sona Gogolkova, Elen Stastna, Michal Zahradnicek, Stefan Savic, Arash Davani, Edward Hulo and Emil Martinka

BioTech 2024, 13(3), 36; https://doi.org/10.3390/biotech13030036 - 19 Sep 2024

Abstract

Chronic wounds result from the body’s inability to heal, causing pain, pathogen entry, limited treatment options, and societal burden. Diabetic foot ulcers are particularly challenging, often leading to severe complications like leg amputation. A clinical study tested AMNIODERM+^®, a new device [...] Read more.

Chronic wounds result from the body’s inability to heal, causing pain, pathogen entry, limited treatment options, and societal burden. Diabetic foot ulcers are particularly challenging, often leading to severe complications like leg amputation. A clinical study tested AMNIODERM+^®, a new device with a lyophilized human amniotic membrane (HAM), on chronic diabetic foot ulcers. Participants had diabetic neuropathic or neuroischemic leg wounds (2–16 cm²) unhealed by 20% after six weeks of standard care. This study showed significant wound healing improvements with AMNIODERM+^®. The median wound size reduction after 12 weeks was 95.5%, far exceeding the null hypothesis of 20% change. Additionally, 65% of patients achieved complete ulceration healing, surpassing the 50% efficacy requirement. The median time to full closure was 11.4 weeks, with the proportion of completely healed patients rising progressively, reaching 55% by week 11. These findings, from the clinical trial “Freeze-dried amniotic membrane in the treatment of nonhealing wounds”, suggest AMNIODERM+^® as a promising future treatment for chronic diabetic foot ulcers. The published results were obtained as part of a clinical trial entitled “Freeze-dried amniotic membrane in the treatment of nonhealing wounds: a single-arm, retrospectively-perspective clinical trial”, EUDAMED Nr. CIV-SK-22-10-041146. Full article

(This article belongs to the Section Medical Biotechnology)

► Show Figures

Figure 1

Journal Description

BioTech

Latest Articles

Journal Menu

Journal Browser

Highly Accessed Articles

Latest Books

E-Mail Alert

News

Topics

Conferences

Special Issues

Further Information

Guidelines

MDPI Initiatives

Follow MDPI