In this work, the effect of moderate electromagnetic fields (2.5, 10, and 15 mT) was studied using an immersed coil inserted directly into a bioreactor on batch cultivation of yeast under both aerobic and anaerobic conditions. Throughout the cultivation, parameters, including CO2 levels, O2 saturation, nitrogen consumption, glucose uptake, ethanol production, and yeast growth (using OD 600 measurements at 1-h intervals), were analysed. The results showed that 10 and 15 mT magnetic fields not only statistically significantly boosted and sped up biomass production (by 38-70%), but also accelerated overall metabolism, accelerating glucose, oxygen, and nitrogen consumption, by 1-2 h. The carbon balance analysis revealed an acceleration in ethanol and glycerol production, albeit with final concentrations by 22-28% lower, with a more pronounced effect in aerobic cultivation. These findings suggest that magnetic fields shift the metabolic balance toward biomass formation rather than ethanol production, showcasing their potential to modulate yeast metabolism. Considering coil heating, opting for the 10 mT magnetic field is preferable due to its lower heat generation. In these terms, we propose that magnetic field can be used as novel tool to increase biomass yield and accelerate yeast metabolism.
- Klíčová slova
- Aerobic, Anaerobic, Batch fermentation, Biomass, Magnetic field, Metabolism acceleration, Yeast,
- MeSH
- aerobióza MeSH
- anaerobióza MeSH
- biomasa * MeSH
- bioreaktory mikrobiologie MeSH
- dusík metabolismus MeSH
- ethanol * metabolismus MeSH
- fermentace * MeSH
- glukosa metabolismus MeSH
- glycerol metabolismus MeSH
- kyslík metabolismus MeSH
- magnetické pole * MeSH
- Saccharomyces cerevisiae * metabolismus růst a vývoj MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- dusík MeSH
- ethanol * MeSH
- glukosa MeSH
- glycerol MeSH
- kyslík MeSH
BACKGROUND: Obesity and related metabolic diseases are becoming a worldwide epidemic, leading to increased mortality and heavy medical costs. Our Chinese herbal formula Xiao-Gao-Jiang-Zhuo (XGJZ) has remarkable effects on curing obese patients in the clinic, but the cellular and molecular basis remains unknown. This study aimed to reveal the molecular mechanism involved in adipogenesis in vitro. METHODS: Chinese herbal formula XGJZ-containing serum was prepared from XGJZ-treated obesity model rats. The function of XGJZ-containing serum was validated in 3T3-L1 preadipocytes. Oil O staining was performed to determine intracellular lipid accumulation in differentiated 3T3-L1 cells. The expression of pro-adipogenic transcription factors was measured to further validate the adipogenesis of 3T3-L1 adipocytes. The contents of triglyceride (TG), free fatty acid (FFA), and glycerin, along with the activities of lipid metabolism-related enzymes (including FAT, FATP1, DGAT, GPAT, ATGL, and HSL) were measured to study the lipogenesis in 3T3-L1 adipocytes. RESULTS: XGJZ-containing serum inhibited 3T3-L1 differentiation, decreased intracellular lipid accumulation, and suppressed the expression of pro-adipogenic transcription factors in differentiated 3T3-L1 cells. The contents of TG, FFA, and glycerin were decreased when treated with XGJZ-containing serum, which also modulated lipid metabolism-related enzyme activities. The activities of fatty acid transporters (FAT, FATP1) and lipid mobilization enzymes (ATGL, HSL) were promoted, while activities of triglyceride biosynthesis enzymes (DGAT, GPAT) were attenuated in differentiated 3T3-L1 cells. CONCLUSION: XGJZ-containing serum has inhibitory effects on adipogenesis in 3T3-L1 preadipocytes, affirming the effect of XGJZ in treating obesity. It provides evidence for the mechanism of obesity.
- Klíčová slova
- 3T3-L1 preadipocytes, XGJZ, adipokines, lipid accumulation,
- MeSH
- adipogeneze * MeSH
- buňky 3T3-L1 MeSH
- glycerol * metabolismus farmakologie MeSH
- krysa rodu Rattus MeSH
- lidé MeSH
- myši MeSH
- obezita MeSH
- PPAR gama metabolismus MeSH
- transkripční faktory MeSH
- triglyceridy MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- glycerol * MeSH
- PPAR gama MeSH
- transkripční faktory MeSH
- triglyceridy MeSH
Trypanosoma brucei is a causative agent of the Human and Animal African Trypanosomiases. The mammalian stage parasites infect various tissues and organs including the bloodstream, central nervous system, skin, adipose tissue and lungs. They rely on ATP produced in glycolysis, consuming large amounts of glucose, which is readily available in the mammalian host. In addition to glucose, glycerol can also be used as a source of carbon and ATP and as a substrate for gluconeogenesis. However, the physiological relevance of glycerol-fed gluconeogenesis for the mammalian-infective life cycle forms remains elusive. To demonstrate its (in)dispensability, first we must identify the enzyme(s) of the pathway. Loss of the canonical gluconeogenic enzyme, fructose-1,6-bisphosphatase, does not abolish the process hence at least one other enzyme must participate in gluconeogenesis in trypanosomes. Using a combination of CRISPR/Cas9 gene editing and RNA interference, we generated mutants for four enzymes potentially capable of contributing to gluconeogenesis: fructose-1,6-bisphoshatase, sedoheptulose-1,7-bisphosphatase, phosphofructokinase and transaldolase, alone or in various combinations. Metabolomic analyses revealed that flux through gluconeogenesis was maintained irrespective of which of these genes were lost. Our data render unlikely a previously hypothesised role of a reverse phosphofructokinase reaction in gluconeogenesis and preclude the participation of a novel biochemical pathway involving transaldolase in the process. The sustained metabolic flux in gluconeogenesis in our mutants, including a triple-null strain, indicates the presence of a unique enzyme participating in gluconeogenesis. Additionally, the data provide new insights into gluconeogenesis and the pentose phosphate pathway, and improve the current understanding of carbon metabolism of the mammalian-infective stages of T. brucei.
- MeSH
- adenosintrifosfát metabolismus MeSH
- fosfofruktokinasy metabolismus MeSH
- glukoneogeneze * genetika MeSH
- glukosa metabolismus MeSH
- glycerol metabolismus MeSH
- lidé MeSH
- savci MeSH
- transaldolasa metabolismus MeSH
- Trypanosoma brucei brucei * genetika metabolismus MeSH
- uhlík metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- adenosintrifosfát MeSH
- fosfofruktokinasy MeSH
- glukosa MeSH
- glycerol MeSH
- transaldolasa MeSH
- uhlík MeSH
Chlorinated ethenes (CEs) are common and persistent contaminants of soil and groundwater. Their degradation is mostly driven by a process of bacterial reductive dechlorination (also called organohalide respiration) in anaerobic conditions. This study summarizes the outcomes of the long-term in-situ application of glycerol for the enhanced reductive dechlorination of CEs on a highly contaminated site. Glycerol injection resulted in an almost immediate increase in the abundance of fermentative Firmicutes, which produce essential sources of carbon (acetate) and electrons (H2) for organohalide-respiring bacteria (OHRB) and change groundwater conditions to be suitable for OHRB growth. The decreased redox potential of groundwater promoted also the proliferation of sulfate-reducing bacteria, which compete for electron donors with OHRB but at the same time support their growth by producing essential corrinoids and acetate. A considerable increase in the abundance of OHRB Dehalococcoides, concurrently with vinyl chloride (VC) reductase gene levels, was revealed by real time polymerase chain reaction (qPCR) method. Consistent with the shifts in bacterial populations, the concentrations of pollutants tetrachloroethylene and trichloroethylene decreased during the monitoring period, with rising levels of cis-1,2-dichloroethylene, VC, and most importantly, the final CE degradation products: ethene and ethane. Our study implies the importance of syntrophic bacterial interactions for successful and complete CE degradation and evaluates glycerol as convenient substrate to enhance reductive dechlorination and as an effective source of electrons for OHRB.
- Klíčová slova
- Bioremediation, Chlorinated ethenes, Enhanced reductive dechlorination, Glycerol, Organohalide-respiring bacteria,
- MeSH
- Bacteria genetika metabolismus MeSH
- biodegradace MeSH
- chemické látky znečišťující vodu * metabolismus MeSH
- Chloroflexi * metabolismus MeSH
- glycerol metabolismus MeSH
- korinoidy * MeSH
- mikrobiota * MeSH
- oxidoreduktasy metabolismus MeSH
- půda MeSH
- sírany metabolismus MeSH
- tetrachlorethylen * metabolismus MeSH
- trichlorethylen * metabolismus MeSH
- uhlík metabolismus MeSH
- vinylchlorid * MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- chemické látky znečišťující vodu * MeSH
- ethylene MeSH Prohlížeč
- glycerol MeSH
- korinoidy * MeSH
- oxidoreduktasy MeSH
- půda MeSH
- sírany MeSH
- tetrachlorethylen * MeSH
- trichlorethylen * MeSH
- uhlík MeSH
- vinylchlorid * MeSH
Strains of Limosilactobacillus reuteri are used as starter and bioprotective cultures and contribute to the preservation of food through the production of fermentation metabolites lactic and acetic acid, and of the antimicrobial reuterin. Reuterin consists of acrolein and 3-hydroxypropionaldehyde (3-HPA), which can be further metabolized to 1,3-propanediol and 3-hydroxypropionic acid (3-HP). While reuterin has been the focus of many investigations, the contribution of 3-HP to the antimicrobial activity of food related reuterin-producers is unknown. We show that the antibacterial activity of 3-HP was stronger at pH 4.8 compared to pH 5.5 and 6.6. Gram-positive bacteria were in general more resistant against 3-HP and propionic acid than Gram-negative indicator strains including common food pathogens, while spoilage yeast and molds were not inhibited by ≤ 640 mM 3-HP. The presence of acrolein decreased the minimal inhibitory activity of 3-HP against E. coli indicating synergistic antibacterial activity. 3-HP was formed during the growth of the reuterin-producers, and by resting cells of L. reuteri DSM 20016. Taken together, this study shows that food-related reuterin producers strains synthesize a second antibacterial compound, which might be of relevance when strains are added as starter or bioprotective cultures to food products.
- Klíčová slova
- 3-Hydroxypropionic acid, Antibacterial, Antifungal, Limosilactobacillus reuteri, Reuterin,
- MeSH
- antiinfekční látky chemie metabolismus farmakologie MeSH
- Bacteria účinky léků růst a vývoj MeSH
- fermentace MeSH
- glyceraldehyd analogy a deriváty chemie metabolismus MeSH
- glycerol metabolismus MeSH
- koncentrace vodíkových iontů MeSH
- kyselina mléčná analogy a deriváty chemie metabolismus farmakologie MeSH
- kyselina octová metabolismus MeSH
- Lactobacillaceae chemie růst a vývoj metabolismus MeSH
- potravinářská mikrobiologie MeSH
- propan chemie metabolismus MeSH
- stabilita léku MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- 3-hydroxypropionaldehyde MeSH Prohlížeč
- antiinfekční látky MeSH
- glyceraldehyd MeSH
- glycerol MeSH
- hydracrylic acid MeSH Prohlížeč
- kyselina mléčná MeSH
- kyselina octová MeSH
- propan MeSH
BACKGROUND: Inadequate blood supply is one of the major risk factors for anastomotic leak after low anterior rectal resection. Early detection of ischemia might predict complicated healing and enable anastomotic salvage, which is associated with better outcomes. A microdialysis offers a real-time evaluation of adequate bowel perfusion through monitoring of tissue metabolism. In this experimental study, we assessed the role of microdialysis in detecting early ischemia after colorectal anastomosis. MATERIALS AND METHODS: Colorectal anastomosis was performed in six miniature pigs. A microdialysis catheter was placed on the bowel serosa 5 mm proximal to the anastomosis. Metabolic changes were monitored hourly before and after initiating ischemia, which was induced by ligation of the inferior mesenteric artery and skeletonization of the mesocolon. RESULTS: Significant increase in tissue levels of lactate was detected after 60 min of ischemia (13.6 [10.4-16.1] versus 6.75 [1.8-9.6] mmol/L at baseline; P < 0.005). The lactate/pyruvate ratio increased accordingly. The concentration of glycerol increased significantly after 2 hours-from a baseline value of 29.5 (3-84) to 125 (79-201) mmol/L (P < 0.005). The decrease in glucose levels was also significant after 60 minutes-0 (0-0.2) versus 4.7 (3.3-6.8) mmol/L at baseline. However, its values started to decline before ischemia. CONCLUSIONS: Surface microdialysis can detect ischemic changes early and may be a promising method in postoperative monitoring of colorectal anastomosis.
- Klíčová slova
- Anastomotic leak, Anterior resection, Microdialysis,
- MeSH
- anastomóza chirurgická škodlivé účinky MeSH
- glycerol metabolismus MeSH
- ischemie diagnóza etiologie metabolismus MeSH
- krevní glukóza analýza MeSH
- kyselina mléčná metabolismus MeSH
- mikrodialýza metody MeSH
- miniaturní prasata MeSH
- pooperační komplikace diagnóza etiologie metabolismus MeSH
- prasata MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
- Názvy látek
- glycerol MeSH
- krevní glukóza MeSH
- kyselina mléčná MeSH
The aim of the study was to screen Yarrowia lipolytica strains for keto acid production and determine optimal conditions for pyruvic acid biosynthesis from glycerol by the best producer. The analyzed parameters were thiamine concentration, medium pH, stirring speed, and substrate concentration. The screening was performed in flask cultures, whereas pyruvic acid production was carried out in 5-L stirred-tank reactor with 2 L of working volume. In total, 24 Y. lipolytica strains were compared for their abilities to produce pyruvic and α-ketoglutaric acids. The total concentration of both acids ranged from 0.1 to 15.03 g/L. Ten strains were selected for keto acid biosynthesis in bioreactor. The Y. lipolytica SKO 6 strain was identified as the best producer of pyruvic acid. In the selected conditions (thiamine concentration 1.5 μg/L, pH 4.0, stirring speed 800 rpm, 150 g/L of glycerol), the strain Y. lipolytica SKO 6 produced 99.3 g/L of pyruvic acid, with process yield of 0.63 g/g and volumetric production rate of 1.18 g/L/h. Higher titer of pyruvic acid was obtained during fed-batch culture with 200 g/L of glycerol, reaching 125.8 g/L from pure glycerol (yield 0.68 g/g) and 124.4 g/L from crude glycerol (yield 0.62 g/g). Results obtained for the strain Y. lipolytica SKO 6 proved the suitability of microbial production of pyruvic acid at industrial scale.
- MeSH
- bioreaktory MeSH
- glycerol analýza metabolismus MeSH
- kultivační média chemie MeSH
- kyselina pyrohroznová analýza metabolismus MeSH
- kyseliny ketoglutarové analýza metabolismus MeSH
- techniky vsádkové kultivace MeSH
- thiamin analýza MeSH
- Yarrowia růst a vývoj metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- glycerol MeSH
- kultivační média MeSH
- kyselina pyrohroznová MeSH
- kyseliny ketoglutarové MeSH
- thiamin MeSH
Saccharomyces kudriavzevii is a nonconventional and rather osmosensitive yeast with a high potential of use in fermentation processes. To elucidate the basis of its relative osmosensitivity, the role of the STL1 gene encoding a putative glycerol uptake system was studied. Under higher osmotic pressure, the addition of a low amount of glycerol to the growth medium improved the growth of S. kudriavzevii and the expression of the STL1 gene was highly induced. Deletion of this gene decreased the strain's ability to grow in the presence of higher concentrations of salts and other solutes. Moreover, the mutant had a disturbed homeostasis of intracellular pH. Expression of the SkSTL1 gene in Saccharomyces cerevisiae complemented the osmosensitivity of the S. cerevisiae hog1Δ stl1Δ mutant, and the gene's tagging with GFP localized its product to the plasma membrane. Altogether, a deficiency in glycerol uptake did not seem to be the reason for S. kudriavzevii's low osmotolerance; its Stl1 transporter properly contributes to the regulation of intracellular pH and is crucial to its survival of osmotic stress. SIGNIFICANCE AND IMPACT OF THE STUDY: An increasing demand for food products with benefits for human health turns the attention to less-exploited nonconventional yeasts with interesting traits not found in Saccharomyces cerevisiae. Among them, Saccharomyces kudriavzevii has good potential for aroma-compound production, fermentations and other biotechnological applications, but it is less adapted to stressful industrial conditions. This report studied S. kudriavzevii relative osmosensitivity and its capacity for active glycerol uptake. The results obtained (on the activity and physiological function of S. kudriavzevii glycerol transporter) may contribute to a further engineering of this species aiming to improve its osmotolerance.
- Klíčová slova
- STL1, glycerol uptake, intracellular pH, osmotolerance, yeast,
- MeSH
- biologický transport genetika fyziologie MeSH
- buněčná membrána metabolismus MeSH
- delece genu MeSH
- fermentace MeSH
- fyziologický stres fyziologie MeSH
- glycerol metabolismus MeSH
- membránové transportní proteiny genetika MeSH
- mitogenem aktivované proteinkinasy genetika MeSH
- osmotický tlak fyziologie MeSH
- Saccharomyces cerevisiae - proteiny genetika MeSH
- Saccharomyces cerevisiae klasifikace genetika metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- glycerol MeSH
- HOG1 protein, S cerevisiae MeSH Prohlížeč
- membránové transportní proteiny MeSH
- mitogenem aktivované proteinkinasy MeSH
- Saccharomyces cerevisiae - proteiny MeSH
- STL1 protein, S cerevisiae MeSH Prohlížeč
S. kudriavzevii has potential for fermentations and other biotechnological applications, but is sensitive to many types of stress. We tried to increase its tolerance and performance via the expression of various transporters from different yeast species. Whereas the overexpression of Z. rouxii fructose uptake systems (ZrFfz1 and ZrFsy1) or a glycerol importer (ZrStl1) did not improve the ability of S. kudriavzevii to consume fructose and survive osmotic stress, the expression of alkali-metal-cation exporters (ScEna1, ScNha1, YlNha2) improved S. kudriavzevii salt tolerance, and that of ScNha1 also the fermentation performance. The level of improvement depended on the type and activity of the transporter suggesting that the natural sensitivity of S. kudriavzevii cells to salts is based on a non-optimal functioning of its own transporters.
- Klíčová slova
- Alkali-metal-cation exporter, Ena ATPase, Fermentation, Nha1 antiporter, Non-conventional yeasts, Salt tolerance,
- MeSH
- fermentace MeSH
- fruktosa metabolismus MeSH
- glycerol metabolismus MeSH
- iontový transport fyziologie MeSH
- osmotický tlak fyziologie MeSH
- proteiny přenášející kationty genetika metabolismus MeSH
- Saccharomyces metabolismus MeSH
- sušené kvasnice MeSH
- tolerance k soli fyziologie MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- fruktosa MeSH
- glycerol MeSH
- proteiny přenášející kationty MeSH
Dekkera bruxellensis is important for lambic beer fermentation but is considered a spoilage yeast in wine fermentation. We compared two D. bruxellensis strains isolated from wine and found that they differ in some basic properties, including osmotolerance. The genomes of both strains contain two highly similar copies of genes encoding putative glycerol-proton symporters from the STL family that are important for yeast osmotolerance. Cloning of the two DbSTL genes and their expression in suitable osmosensitive Saccharomyces cerevisiae mutants revealed that both identified genes encode functional glycerol uptake systems, but only DbStl2 has the capacity to improve the osmotolerance of S. cerevisiae cells.
- MeSH
- Dekkera genetika izolace a purifikace metabolismus fyziologie MeSH
- druhová specificita MeSH
- fungální proteiny genetika metabolismus MeSH
- genom bakteriální genetika MeSH
- glycerol metabolismus MeSH
- osmoregulace genetika MeSH
- protony MeSH
- rekombinantní proteiny genetika metabolismus MeSH
- Saccharomyces cerevisiae genetika MeSH
- symportéry genetika metabolismus MeSH
- testy genetické komplementace MeSH
- víno mikrobiologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Názvy látek
- fungální proteiny MeSH
- glycerol MeSH
- protony MeSH
- rekombinantní proteiny MeSH
- symportéry MeSH
