Microbial transglutaminase (MTG) is an enzyme widely used in the food industry because it creates cross-links between proteins, enhancing the texture and stability of food products. Its unique properties make it a valuable tool for modifying the functional characteristics of proteins, significantly impacting the quality and innovation of food products. In this study, response surface methodology was employed to optimize the fermentation conditions for microbial transglutaminase production by the strain Streptoverticillium cinnamoneum KKP 1658. The effects of nitrogen dose, cultivation time, and initial pH on the activity of the produced transglutaminase were investigated. The significance of the examined factors was determined as follows: cultivation time > nitrogen dose > pH. The interaction between nitrogen dose and cultivation time was found to be crucial, having the second most significant impact on transglutaminase activity. Optimal conditions were identified as 48 h of cultivation with a 2% nitrogen source dose and an initial medium pH of approximately 6.0. Under these conditions, transglutaminase activity ranged from 4.5 to 5.5 U/mL. The results of this study demonstrated that response surface methodology is a promising approach for optimizing microbial transglutaminase production. Future applications of transglutaminase include the development of modern food products with improved texture and nutritional value, as well as its potential use in regenerative medicine for creating biomaterials and tissue scaffolds. This topic is particularly important and timely as it addresses the growing demand for innovative and sustainable solutions in the food and biomedical industries, contributing to an improved quality of life.
Yeasts are unicellular fungi that occur in a wide range of ecological niches, where they perform numerous functions. Furthermore, these microorganisms are used in industrial processes, food production, and bioremediation. Understanding the physiological and adaptive characteristics of yeasts is of great importance from ecological, biotechnological, and industrial perspectives. In this context, we evaluated the abilities to assimilate and ferment different carbon sources, to produce extracellular hydrolytic enzymes, and to tolerate salt stress, heavy metal stress, and UV-C radiation of two isolates of Eremothecium coryli, isolated from Momordica indica fruits. The two isolates were molecularly identified based on sequencing of the 18S-ITS1-5.8S-ITS2 region. Our isolates were able to assimilate nine carbon sources (dextrose, galactose, mannose, cellobiose, lactose, maltose, sucrose, melezitose, and pectin) and ferment three (glucose, maltose, and sucrose). The highest values of cellular dry weight were observed in the sugars maltose, sucrose, and melezitose. We observed the presence of hyphae and pseudohyphae in all assimilated carbon sources. The two isolates were also capable of producing amylase, catalase, pectinase, and proteases, with the highest values of enzymatic activity found in amylase. Furthermore, the two isolates were able to grow in media supplemented with copper, iron, manganese, nickel, and zinc and to tolerate saline stress in media supplemented with 5% NaCl. However, we observed a decrease in CFU at higher concentrations of these metals and NaCl. We also observed morphological changes in the presence of metals, which include changes in cell shape and cellular dimorphisms. The isolates were sensitive to UV-C radiation in the shortest exposure time (1 min). Our findings reinforce the importance of endophytic yeasts for biotechnological and industrial applications and also help to understand how these microorganisms respond to environmental variations caused by human activities.
- MeSH
- endofyty * izolace a purifikace genetika metabolismus fyziologie klasifikace účinky záření MeSH
- fermentace MeSH
- fylogeneze MeSH
- fyziologický stres * MeSH
- metabolismus sacharidů * MeSH
- ovoce * mikrobiologie MeSH
- Saccharomycetales * izolace a purifikace genetika fyziologie metabolismus účinky záření klasifikace MeSH
- těžké kovy toxicita MeSH
- ultrafialové záření MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: The medication used to treat benign prostate hyperplasia (BPH), a common condition in men over 50 years of age, can alter the levels of biomarkers used in prostate cancer detection. Commonly used medications for BPH include alpha-blockers, 5-alpha reductase inhibitors (5-ARIs), and muscarinic antagonists. We studied the impact of these drugs on total prostate-specific antigen (tPSA), free PSA (fPSA), [-2]proPSA, fPSA/tPSA ratio, and the Prostate Health Index (PHI), as well as novel potential biomarkers in the form of glycan composition of fPSA. PATIENTS AND METHODS: Serum samples were collected from 564 males with BPH, with a mean age of 68.5 years. The samples were used to measure levels of tPSA, fPSA, and [-2]proPSA. The fPSA/tPSA and PHI were then calculated. The glycan composition of fPSA was analyzed using lectin-based glycoprofiling. Pharmacotherapy data was collected from the patients' medical records. RESULTS: Alpha-blocker monotherapy was associated with higher fPSA and fPSA/tPSA ratio, and decreased PHI. Levels of tPSA were not impacted. Alpha-blocker and 5-ARI dual therapy was associated with reduced levels of fPSA, [-2]proPSA, and PHI. Therapy combining alpha-blockers and antimuscarinic agents did not significantly influence biomarker levels apart from an increase in a Maackia amurensis lectin-recognized glycan originating in fPSA. CONCLUSION: BPH pharmacotherapy notably affects prostate cancer biomarkers. Recognizing the impact of pharmacotherapy is crucial for achieving an accurate diagnosis of prostate cancer and for planning treatment.
- MeSH
- alfa blokátory terapeutické užití MeSH
- antagonisté muskarinových receptorů * terapeutické užití MeSH
- glykosylace MeSH
- hyperplazie prostaty * krev farmakoterapie MeSH
- inhibitory 5-alfa-reduktasy terapeutické užití MeSH
- lidé středního věku MeSH
- lidé MeSH
- nádory prostaty krev farmakoterapie MeSH
- prostata patologie metabolismus MeSH
- prostatický specifický antigen * krev MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- Publikační typ
- časopisecké články MeSH
Feruloyl esterases (FAEs) are a crucial component of the hemicellulose-degrading enzyme family that facilitates the degradation of lignocellulose while releasing hydroxycinnamic acids such as ferulic acid with high added value. Currently, the low enzyme yield of FAEs is one of the primary factors limiting its application. Therefore, in this paper, we optimized the fermentation conditions for the expression of FAE BpFaeT132C-D143C with excellent thermal stability in Escherichia coli by experimental design. Firstly, we explored the effects of 11 factors such as medium type, isopropyl-β-D-thiogalactopyranoside (IPTG) concentration, and inoculum size on BpFaeT132C-D143C activity separately by the single factor design. Then, the significance of the effects of seven factors, such as post-induction temperature, shaker rotational speed, and inoculum size on BpFaeT132C-D143C activity, was analyzed by Plackett-Burman design. We identified the main factors affecting the fermentation conditions of E. coli expressing BpFaeT132C-D143C as post-induction temperature, pre-induction period, and post-induction period. Finally, we used the steepest ascent path design and response surface method to optimize the levels of these three factors further. Under the optimal conditions, the activity of BpFaeT132C-D143C was 3.58 U/ml, which was a significant 6.6-fold increase compared to the pre-optimization (0.47 U/ml), demonstrating the effectiveness of this optimization process. Moreover, BpFaeT132C-D143C activity was 1.52 U/ml in a 3-l fermenter under the abovementioned optimal conditions. It was determined that the expression of BpFaeT132C-D143C in E. coli was predominantly intracellular in the cytoplasm. This study lays the foundation for further research on BpFaeT132C-D143C in degrading agricultural waste transformation applications.
- MeSH
- Escherichia coli * genetika metabolismus enzymologie MeSH
- fermentace * MeSH
- isopropylthiogalaktosid metabolismus MeSH
- karboxylesterhydrolasy * genetika metabolismus chemie biosyntéza MeSH
- kultivační média chemie MeSH
- kyseliny kumarové metabolismus MeSH
- lignin MeSH
- rekombinantní proteiny genetika metabolismus biosyntéza chemie MeSH
- stabilita enzymů MeSH
- teplota MeSH
- Publikační typ
- časopisecké články MeSH
In this study, lactic acid bacteria (LAB) isolation from fermented foods and molecular identification using magnetic bead technology were performed. And then exopolysaccharide (EPS) production possibility was tested in agar medium, and the positive ones were selected for the next step. The bacteria that could produce higher carbohydrate level were grown in MRS medium fortified with whey and pumpkin waste. In our study, 19 different LAB species were identified from fermented products collected from different places in Hatay (Türkiye) province. In molecular identification, universal primer pairs, p806R/p8FPL, and PEU7/DG74 were used for PCR amplification. After that, PCR products purified using paramagnetic bead technology were sequenced by the Sanger sequencing method. The dominant species, 23.8% of the isolates, were identified as Lactiplantibacillus plantarum. As a technological property of LAB, exopolysaccharide production capability of forty-two LAB isolate was tested in agar medium, and after eleven isolates were selected as positive. Two LAB (Latilactobacillus curvatus SHA2-3B and Loigolactobacillus coryniformis SHA6-3B) had higher EPS production capability when they were grown in MRS broth fortified with pumpkin waste and whey. The highest EPS content (1750 mg/L glucose equivalent) was determined in Loigolactobacillus coryniformis SHA6-3B grown in MRS broth fortified with 10% pumpkin waste. Besides the produced EPS samples were validated with FTIR and SEM methods.
- MeSH
- bakteriální polysacharidy * biosyntéza metabolismus MeSH
- Cucurbita mikrobiologie MeSH
- fermentace MeSH
- fermentované potraviny * mikrobiologie MeSH
- fylogeneze MeSH
- kultivační média chemie MeSH
- Lactobacillales * izolace a purifikace klasifikace genetika metabolismus MeSH
- odpadní produkty * analýza MeSH
- potravinářská mikrobiologie * MeSH
- RNA ribozomální 16S genetika MeSH
- syrovátka MeSH
- Publikační typ
- časopisecké články MeSH
Ultraviolet (UV) radiation is a significant environmental stressor that affects the growth, physiology, and biochemical integrity of various organisms. This study investigates the potential protective effects of a zinc-cysteine (Zn-Cys) complex against UV-C radiation, with a focus on its impact on selected microalgae (Coccomyxa peltigerae and Parachlorella kessleri) and maize (Zea mays L.). We demonstrate that exposure of the Zn-Cys complex to UV-C (254 nm) results in the formation of fluorescent photoproducts, which exhibit UV-protective properties. The study reveals that Zn-Cys significantly mitigates UV-induced stress. In both microalgae species, the Zn-Cys complex enhanced growth even under UV exposure, with the 20% concentration showing the most robust protective effects. Further hyperspectral imaging confirmed the protective mechanism of Zn-Cys by monitoring changes in light reflectance in Parachlorella kessleri, indicating reduced photosynthetic efficiency and structural alterations induced by UV exposure, while Zn-Cys significantly mitigated these effects. In addition, in maize plants (Zea mays L.), Zn-Cys treatment preserved chlorophyll content and reduced polyphenol accumulation, indicating reduced oxidative stress. These findings highlight the potential of the Zn-Cys complex as a sustainable and cost-effective strategy for UV protection in both terrestrial and extraterrestrial agriculture, advancing our understanding of plant adaptation to extreme environments.
- MeSH
- Chlorophyta účinky záření účinky léků MeSH
- cystein * chemie farmakologie MeSH
- fotosyntéza účinky léků účinky záření MeSH
- komplexní sloučeniny * chemie farmakologie MeSH
- kukuřice setá účinky záření účinky léků metabolismus MeSH
- mikrořasy účinky záření účinky léků MeSH
- ultrafialové záření * MeSH
- zinek * chemie farmakologie MeSH
- Publikační typ
- časopisecké články MeSH
Numerous studies have reported that increased interleukin 6 (IL-6) and soluble IL-6 receptor (sIL-6) levels induce inflammatory conditions. However, the exact mechanisms by which IL-6 drives inflammatory conditions remain unclear. Therefore, we investigated the potential role of IL-6/sIL-6R in inducing energy metabolism, including glycolysis, oxidative phosphorylation, lactate secretion and Akt/mTOR phosphorylation, in Jurkat cells, and whether IL-6 would increase the risk of developing inflammatory conditions due to the high metabolic profile of the T cells. Jurkat CD4 T-cell lines were stimulated with IL-6/sIL-6R for 24 h prior to 48-h stimulation with anti-CD3/CD28. Lactate secretion, glycolysis and oxidative phosphorylation levels were characterized using the Seahorse XF analyser. The Akt and mTOR phosphorylation status was detected using Western blotting. IL-6/sIL-6R significantly induced glycolysis and oxidative phosphorylation and their related parameters, including glycolytic capacity and maximal respiration, followed by significantly increased lactate secretion. Akt and mTOR phosphorylation were increased, which could have resulted from energy metabolism. Here we show that IL-6 enhanced the metabolic profile of Jurkat cells. This effect could have consequences for the metabolism-related signalling pathways, including Akt and mTOR, suggesting that IL-6 might promote T-cell energy metabolism, where T-cell hyperactivity might increase the inflammatory disease risk. The findings should be validated using studies on primary cells isolated from humans.
- MeSH
- energetický metabolismus * účinky léků MeSH
- fosforylace účinky léků MeSH
- glykolýza účinky léků MeSH
- interleukin-6 * metabolismus MeSH
- Jurkat buňky MeSH
- kyselina mléčná metabolismus MeSH
- lidé MeSH
- oxidativní fosforylace účinky léků MeSH
- protoonkogenní proteiny c-akt * metabolismus MeSH
- signální transdukce * účinky léků MeSH
- TOR serin-threoninkinasy * metabolismus MeSH
- zánět * metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Leukemia is driven by complex interactions within the inherently hypoxic bone marrow microenvironment, impacting both disease progression and therapeutic resistance. Co-cultivation of leukemic cells with feeder cells has emerged as a valuable tool to mimic the bone marrow niche. This study explores the interplay between human commercial SD-1 and patient-derived UPF26K leukemic cell lines with feeders - human fibroblasts (NHDF) and mesenchymal stem cells (hMSCs) under normoxic and hypoxic conditions. RESULTS: Co-cultivation with feeders significantly enhances proliferation and glycolytic activity in the SD-1 cells, improving their viability, while this interaction inhibits the growth and glucose metabolism of the feeders, particularly NHDF. In contrast, UPF26K cells show reduced proliferation when co-cultivated with the feeders while this interaction stimulates NHDF and hMSCs proliferation and glycolysis but reduce their mitochondrial metabolism with hypoxia amplifying these effects. CONCLUSIONS: Cells that switch to glycolysis during co-cultivation, particularly under hypoxia, benefit most from these low oxygen conditions. Due to this leukemic cells' response heterogeneity, targeting microenvironmental interactions and oxygen levels is crucial for personalized leukemia therapy. Advancing co-cultivation models, particularly through innovations like spheroids, can further enhance in vitro studies of primary leukemic cells and support the testing of novel therapies.
- MeSH
- fibroblasty * metabolismus MeSH
- glykolýza MeSH
- hypoxie buňky MeSH
- kokultivační techniky metody MeSH
- leukemie * patologie metabolismus MeSH
- lidé MeSH
- mezenchymální kmenové buňky * metabolismus MeSH
- mitochondrie metabolismus MeSH
- nádorové buněčné linie MeSH
- nádorové mikroprostředí MeSH
- proliferace buněk MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
DC-SIGN, a C-type lectin receptor expressed on immune cells, is considered a promising target for immunomodulatory and antiviral therapies. While mannose-based glycomimetics have been extensively studied as DC-SIGN ligands, fucose-based strategies remain underexplored. This study explores the fucosylation of linear alcohols and sugars using eight different fucosyl donors, aiming at designing strategies for the development of fucose-based glycomimetics targeting DC-SIGN. Four types of leaving groups and two different acyl-based protecting groups on the donors were tested. The glycosylation of 3-azidopropan-1-ol exclusively yielded the β-anomer, demonstrating high stereoselectivity. The azido group in the product is versatile, allowing for direct click chemistry reactions or reduction to an amine for further functionalization. Both types of reactions were demonstrated in a model reaction. In the glycosylation of a sugar, a disaccharide moiety of Lewis X antigen was selected as a target molecule. Only one of the eight tested fucosyl donors worked well in this reaction and provided the product in a reasonable yield. The disaccharide was also equipped with the 3-azidopropyl linker, facilitating future modifications. Finally, NMR studies confirmed compatibility of the linker with canonical Ca2+-dependent carbohydrate binding to DC-SIGN, suggesting potential for further development of fucose-based glycomimetics targeting this C-type lectin receptor.
- MeSH
- fukosa * chemie MeSH
- glykosidy * chemie chemická syntéza farmakologie metabolismus MeSH
- glykosylace MeSH
- lektiny typu C * metabolismus antagonisté a inhibitory MeSH
- lidé MeSH
- molekulární struktura MeSH
- molekuly buněčné adheze * metabolismus antagonisté a inhibitory MeSH
- receptory buněčného povrchu * metabolismus antagonisté a inhibitory MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
The effect of the antibiotic tetracycline on the growth and photosynthetic activity of the diatoms Cyclotella caspia Grunow, 1878, and Thalassiosira weissflogii (Grunow) G.A. Fryxell & Hasle, 1977, has been studied. The rate of tetracycline concentration decreases in the medium with and without has been estimated. The expediency and prospects of using diatoms as promising objects for water purification from tetracycline are shown. It was found that the rate and efficiency of tetracycline removal from the medium in the presence of algae depends on the initial content of the pollutant in the medium. The maximum efficiency of antibiotic removal is observed at concentrations of the pollutant provoking the hormesis growth of algae at 5 mg/L in C. caspia and 10 mg/L in T. weissflogii. In samples with C. caspia and tetracycline, the residual antibiotic content in the medium was 10-14% lower than in samples without algae. In the experiment with T. weissflogii, this indicator reached 15-16%. At concentrations of the pollutant, 2 and 10 mg/L in C. caspia and 2 and 5 mg/L in T. weissflogii, the effectiveness of removing the antibiotic by algae was lower. The residual content of tetracycline in the medium was on average 8-11% lower than in samples without algae. At antibiotic concentrations of 15 and 20 mg/L, the rate of its removal in samples with and without algae did not significantly differ. This result is obviously due to the toxic effect of tetracycline on cells, leading to pronounced inhibition of algae growth and/or death.
