Medium-chain Fatty Acids Research Articles

Solvent‐free crystallization for fractionation of virgin coconut oil: Effect of process conditions on kinetics and crystal properties

AbstractThis research was aimed to conduct solvent‐free crystallization and fractionation of virgin coconut oil (VCO). This study investigated the effect of surface contact area‐to‐volume ratio and mixing conditions on crystal characteristics. Commercially available VCO samples, extracted by different methods, were analyzed for fatty acid and triacylglycerol composition using GC–MS and UPLC–ELSD. VCO extracted through enzyme and chilling–thawing cycles exhibited higher medium chain fatty acids and medium chain triacylglycerols, which was selected for crystallization studies. Isothermal crystallization of VCO at 21°C assessed the effect of scale of crystallizer tube on cooling rate using Newton's law of cooling equation. The smaller tube (20 mm diameter) with a higher surface contact area‐to‐volume ratio produced 4.1‐fold faster cooling and a higher crystallization yield (44%) compared to the 80 mm diameter tube when no agitation was applied. Whereas mixing with a magnetic stirrer produced a large number of nuclei, generating intense crystallization heat, and rendering the crystals inseparable from liquid fraction. The process of VCO crystallization was also studied for kinetics and crystal growth mechanism using the Avrami model employing angled rotation of sample tube during non‐isothermal crystallization. Angled rotary mixing produced stable crystals and significantly increased the crystallization rate (t1/2 = 26.86 min) with polyhedral or spherulitic crystal growth (n = 3.25) compared to non‐mixing condition (t1/2 = 161.87 min) with restricted linear growth (n = 1.64). DSC analysis of VCO and its fractions obtained after crystallization using the 20 mm tube with angled rotary mixing indicated shifts in their thermogram peaks, suggesting differences in triacylglycerol compositions.Practical applicationsVirgin coconut oil (VCO) is well recognized for its health‐beneficial properties, yet its application is limited due to its complex triglyceride composition. The solvent‐free crystallization method facilitates VCO fractionation into phases with different melting points without hazardous solvents. The efficacy of this process is highly dependent on temperature, cooling rate, and agitation, which affects crystal morphology and growth kinetics. This study addresses these challenges in the scarcely investigated area of VCO crystallization. The resulting fractions can be customized for specific applications based on their altered functional properties in producing baking and confectionery coatings, salad dressings, and so on. The olein fraction obtained in this study can be utilized in nutraceutical formulations targeting digestive and cognitive health, as it is reported to be rich in medium chain triglycerides. This research optimizes process conditions, suggesting an efficient method for dry‐fractionating VCO, which is significantly associated with the food and pharmaceutical industries.

Interface-based kinetic model considering the integral stereoselectivity of lipases on tricapryloylglycerol in a reverse micelle system

Lipases are essential enzymes with unique selectivity, making them valuable in industrial applications. Understanding the integral stereoselectivity of lipases during triacylglycerol (TAG) hydrolysis is crucial for producing high-value products, such as structured lipids. This study developed an analytical method and an interface-based kinetic model to determine integral stereoselectivities on tricapryloylglycerol (TCG), a medium-chain TAG. The analytical method used an HPLC system that simultaneously separated TCG and its hydrolysates with resolution factors of >2.4 and relative standard deviation of retention times <0.3 % within 15 min. The interface-based kinetic model was established to determine the integral stereoselectivities according to the characteristics of the reaction system. The model provided better fitting results for TCG and trioleoylglycerol hydrolysis than a previous model, indicating the successful application in both medium- and long-chain TAGs. This study expanded our understanding of integral stereoselectivity and could facilitate the development of various structured lipid syntheses.

Physicochemical Properties, Heavy Metal and Pesticide Contaminations, and Microbial Limit Tests of Cannabis Oral Drops

Cannabis, a medicinal herb renowned for its therapeutic potential, is utilized in the treatment of various ailments, including the stimulation of appetite in cancer patients. In Thailand, the predominant dosage form is oral drops; however, the absence of robust quality control measures for these products poses significant health risks to patients. The primary objectives of this study were to formulate and evaluate cannabis oral drops, with a comprehensive evaluation encompassing physicochemical properties such as appearance, pH, Δ9-tetrahydrocannabinol, and cannabidiol contents. Furthermore, heavy metals (specifically arsenic, cadmium, and lead), pesticides (including pyrethroids, organochlorines, organophosphates, and carbamates), and microbial contaminations (Staphylococcus aureus, Clostridium spp., and Salmonella spp.) were evaluated. The results revealed that the optimal formulation, comprising cannabis extract derived from supercritical carbon dioxide fluid, vitamin E acetate, and medium-chain triglyceride oil, exhibited a pH of approximately 4.0, a Δ9-tetrahydrocannabinol content of approximately 27 mg/mL, and a cannabidiol content of approximately 9 mg/mL. Furthermore, there were no heavy metals or pesticides detected. Microbial limit tests demonstrated the absence of Staphylococcus aureus, Clostridium spp., and Salmonella spp. In conclusion, this endeavor successfully achieved the development of high-quality cannabis oral drops, showcasing the potential for safe and therapeutic use.

Impact of cell voltage on synthesis of caproic acid from carbon dioxide and ethanol in direct current powered microbial electrosynthesis cell

Production of medium chain fatty acids (MCFAs) from CO2 through microbial electrosynthesis (MES) holds great potential. The present study investigated the effect of cathode voltages of − 0.8 V (MES-1), −1.0 V (MES-2) and −1.2 V vs Ag/AgCl (MES-3), on the production of MCFAs from CO2 and ethanol using an enriched culture. Direct current (DC) power supply was used to maintain constant cathode voltages. The highest amounts of caproic acid were produced in MES-2 at an average concentration of 1.51 ± 0.14 g/L with a maximum selectivity of 68 ± 7 %. Microbial diversity analysis showed abundance of the Clostridiaceae family that allowed chain elongation in all MES reactors. This study shows that potentiostatic control approach for MCFA synthesis, can be replaced by DC power supply in future MES setups. Using selective culture enrichment, MES efficiently produces MCFAs from CO2 and ethanol, with −1.0 V yielding the highest caproic acid.

Pharmacological inhibition of ENT1 enhances the impact of specific dietary fats on energy metabolism gene expression

Medium chain fatty acids are commonly consumed as part of diets for endurance sports and as medical treatment in ketogenic diets where these diets regulate energy metabolism and increase adenosine levels. However, the role of the equilibrative nucleoside transporter 1 (ENT1), which is responsible for adenosine transport across membranes in this process, is not well understood. Here, we investigate ENT1 activity in controlling the effects of two dietary medium chain fatty acids (decanoic and octanoic acid), employing the tractable model system Dictyostelium. We show that genetic ablation of three ENT1 orthologues unexpectedly improves cell proliferation specifically following decanoic acid treatment. This effect is not caused by increased adenosine levels triggered by both fatty acids in the presence of ENT1 activity. Instead, we show that decanoic acid increases expression of energy-related genes relevant for fatty acid β-oxidation, and that pharmacological inhibition of ENT1 activity leads to an enhanced effect of decanoic acid to increase expression of tricarboxylicacid cycle and oxidative phosphorylation components. Importantly, similar transcriptional changes have been shown in the rat hippocampus during ketogenic diet treatment. We validated these changes by showing enhanced mitochondria load and reduced lipid droplets. Thus, our data show that ENT1 regulates the medium chain fatty acid-induced increase in cellular adenosine levels and the decanoic acid-induced expression of important metabolic enzymes in energy provision, identifying a key role for ENT1 proteins in metabolic effects of medium chain fatty acids.

Advanced Statistical Optimization for Enhanced Medium-Chain Fatty Acid Production

In recent years, the development of feed ingredients with natural additives has gained significant importance in increasing the health and quality of animal products, as well as in promoting weight gain in animals. Since Salmonella infection is a significant disease that transmits from animals to humans, the inhibition of Salmonella species can be achieved particularly through the improvement of gastrointestinal metabolism in chickens. At this point, the effectiveness of using MCFA (Medium Chain Fatty Acids) as a feed additive has been proven. MCFA are composed of a mixture of various fatty acids, including acetic acid, butyric acid, hexanoic acid, etc. Highest portion of MCFA are hexanoic acid. Besides feed additives hexanoic acid play a crucial role as primary resources in various industries, including the chemical, food, agricultural, and biofuel sectors. It is typically obtained from petrochemical-based solutions but there has been a growing focus on biotechnological production and natural sources in recent years. One of the mostly known bioprocess to produce MCFA is chain elongation (conversion of acetate and ethanol into MCFA by β oxidation reaction) by Clostridium kluyveri. However, as in most biotechnological processes, there are low yields and high costs in these reactions as well. In this study, Box-Behnken Design, a statistical experimental design method, was used to optimize the concentrations of acetate, ethanol (the two primary components of chain elongation reactions) and pH for MCFA production via chain elongation reactions with Clostridium kluyveri. Batch experiments were performed at 30°C and 37°C to also see the effect of temperature. Higher values of hexanoic acid and bacterial growth were observed at 37°C. From an economic perspective, a 14% reduction in costs has been observed with optimized components.

The Cytoprotective Effect of C60 Derivatives in the Self-Microemulsifying Drug Delivery System against Triptolide-Induced Cytotoxicity In Vitro.

The aim of this study was to optimize the formulation of a C60-modified self-microemulsifying drug delivery system loaded with triptolide (C60-SMEDDS/TP) and evaluate the cytoprotective effect of the C60-SMEDDS/TP on normal human cells. The C60-SMEDDS/TP exhibited rapid emulsification, an optimal particle size distribution of 50 ± 0.19 nm (PDI 0.211 ± 0.049), and a near-neutral zeta potential of -1.60 mV. The release kinetics of TP from the C60-SMEDDS/TP exhibited a sustained release profile and followed pseudo-first-order release kinetics. Cellular proliferation and apoptosis analysis indicated that the C60-SMEDDS/TP (with a mass ratio of TP: DSPE-PEG-C60 = 1:10) exhibited lower toxicity towards L02 and GES-1 cells. This was demonstrated by a higher IC50 (40.88 nM on L02 cells and 17.22 nM on GES-1 cells) compared to free TP (21.3 nM and 11.1 nM), and a lower apoptosis rate (20.8% on L02 cells and 26.3% on GES-1 cells, respectively) compared to free TP (50.5% and 47.0%) at a concentration of 50 nM. In comparison to the free TP group, L02 cells and GES-1 cells exposed to the C60-SMEDDS/TP exhibited a significant decrease in intracellular ROS and an increase in mitochondrial membrane potential (ΔψM). On the other hand, the C60-SMEDDS/TP demonstrated a similar inhibitory effect on BEL-7402 cells (IC50 = 28.9 nM) and HepG2 cells (IC50 = 107.6 nM), comparable to that of the free TP (27.2 nM and 90.4 nM). The C60-SMEDDS/TP group also exhibited a similar intracellular level of ROS and mitochondrial membrane potential compared to the SMEDDS/TP and free TP groups. Fullerenol-Grafted Distearoyl Phosphatidylethanolamine-Polyethylene Glycol (DSPE-PEG-C60) was synthesized and applied in the self-microemulsifying drug delivery system. The C60-SMEDDS/TP was formulated using Cremophor EL, medium-chain triglycerides (MCT), PEG-400, and DSPE-PEG-C60, and loaded with triptolide (TP). The toxicity and bioactivity of the C60-SMEDDS/TP were assessed using normal human liver cell lines (L02 cells), normal human gastric mucosal epithelial cell lines (GES-1 cells), and liver cancer cell lines (BEL-7402 cells and HepG2 cells). The production of reactive oxygen species (ROS) after the C60-SMEDDS/TP treatment was assessed using 2',7'-dichlorofluorescein diacetate (DCFDA) staining. The alterations in mitochondrial membrane potential (ΔψM) were assessed by measuring JC-1 fluorescence. The cytoprotection provided by the C60-SMEDDS/TP favored normal cells (L02 and GES-1) over tumor cells (BEL-7402 and HepG2 cells) in vitro. This suggests a promising approach for the safe and effective treatment of TP.

Effect of oral metformin on gut microbiota characteristics and metabolite fractions in normal-weight type 2 diabetic mellitus patients.

To analyze the effect of oral metformin on changes in gut microbiota characteristics and metabolite composition in normal weight type 2 diabetic patients. T2DM patients in the cross-sectional study were given metformin for 12 weeks. Patients with unmedicated T2DM were used as a control group to observe the metrics of T2DM patients treated with metformin regimen. 16S rDNA high-throughput gene sequencing of fecal gut microbiota of the study subjects was performed by llumina NovaSeq6000 platform. Targeted macro-metabolomics was performed on 14 cases of each of the gut microbiota metabolites of the study subjects using UPLC-MS/MS technology. Correlations between the characteristics of the gut microbiota and its metabolites, basic human parameters, glycolipid metabolism indicators, and inflammatory factors were analyzed using spearman analysis. Glycolipid metabolism indexes and inflammatory factors were higher in normal-weight T2DM patients than in the healthy population (P<0.05), but body weight, BMI, waist circumference, and inflammatory factor concentrations were lower in normal-weight T2DM patients than in obese T2DM patients (P<0.05). Treatment with metformin in T2DM patients improved glycolipid metabolism, but the recovery of glycolipid metabolism was more pronounced in obese T2DM patients. None of the differences in α-diversity indexes were statistically significant (P>0.05), and the differences in β-diversity were statistically significant (P <0.05). Community diversity and species richness recovered after metformin intervention compared to before, and were closer to the healthy population. We found that Anaerostipes/Xylose/Ribulose/Xylulose may play an important role in the treatment of normal-weight T2DM with metformin by improving glycemic lipids and reducing inflammation. And Metformin may play a role in obese T2DM through Romboutsia, medium-chain fatty acids (octanoic acid, decanoic acid, and dodecanoic acid). Gut microbial dysbiosis and metabolic disorders were closely related to glucose-lipid metabolism and systemic inflammatory response in normal-weight T2DM patients. Metformin treatment improved glucose metabolism levels, systemic inflammation levels in T2DM patients, closer to the state of healthy population. This effect may be mediated by influencing the gut microbiota and microbial host co-metabolites, mainly associated with Anaerostipes and xylose/Ribulose/Xylulose. Metformin may exert its effects through different pathways in normal-weight versus obese T2DM patients.

Two PNPLA2 heterozygous mutations result in neutral lipid storage disease with myopathy: a case report

BackgroundNeutral Lipid Storage Disease with Myopathy (NLSDM) is a rare lipid metabolism disorder caused by PNPLA2 gene mutations. Clinical manifestations are heterogeneous, and diagnosis is often delayed, usually gaining patients’ attention due to the increased risk of cardiomyopathy.Case presentationWe herein report a 36-year-old Asian male presenting with progressive limb weakness, muscle atrophy of limbs and trunk, dysarthria, and heart failure. Electromyography indicated myogenic changes, and muscle biopsy results revealed characteristics of lipid storage myopathy. Genetic analysis of PNPLA2 revealed two heterozygous mutations: c.757 + 1G > T (chr11-823588, splice-5) on intron 6 and c.919delG (chr11-823854, p.A307Pfs*13) on exon 7. The patient improved limb strength, and dysarthria disappeared after the Medium Chain Fatty Acids diet.ConclusionsIn conclusion, we report for the first time that the two heterozygous mutations PNPLA2 c.919delG and c.757 + 1G > T together induced NLSDM, which was confirmed by muscle biopsy.

Metabolism Capacity of Medium Chain Fatty Acids from Virgin Coconut Oil: an In Vivo Study

In this study, the types of fatty acids in the liver tissue of mice were evaluated under different diets. As a result, mice fed with a high-fat diet – including basic pellets mixed with eight percent (w/w) peanut oil, two percent (w/w) cholesterol, and 0.1% (w/w) calcium carbonate – showed the highest cholesterol content (1.5 ± 0.08 mg/100 g) compared to normal mice (0.23 ± 0.04 mg/100 g). Whereas the cholesterol content in the liver tissue of mice fed with basic pellets and orally treated with a free fatty acid fraction (FFA1) from virgin coconut oil, which was made up of the C8:0–C12:0 fraction (97.3%) was prepared did not differ from the control group (using only standard pellets). This demonstrated that MCFA (C8:0–C12:0) was directly consumed and converted to energy without accumulating cholesterol. The gas chromatography analysis result showed that the fat droplets in the liver tissues of treated mice were mostly saturated long-chain fatty acids (LCFA), in which C16:0 and C18:0 carbon chain length fatty acids were predominant in the liver tissue. The results confirmed the hypothesis that MCFA is preferentially absorbed and converted to energy while saturated LCFA has a tendency to induce fat accumulation in the metabolic pathway.

Carbon-13-isotopomics and metabolomics of fatty acids from triacylglycerols: overcoming the limitations of GC-C-IRMS for short- and medium-acyl chains.

Carbon-13 isotopomics of triacylglycerol (TAG) fatty acids or free fatty acids in biological matrices holds considerable potential in food authentication, forensic investigations, metabolic studies, and medical research. However, challenges arise in the isotopic analysis of short- and medium-chain (C4 to C10) fatty acid methyl esters (SMCFAMEs) through gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS). The high volatility of these esters results in losses during their preparation, leading to isotopic fractionation. Moreover, the methoxy group added to acyl chains requires the correction of δ13C values, thereby increasing the uncertainty of the final results. Analyzing free fatty acids (FFAs) addresses both issues encountered with SMCFAMEs. To achieve this objective, we have developed a new protocol enabling the isotopomics of individual fatty acids (FAs) by GC-C-IRMS. The same experiment also provides the FA profile, i.e., the relative percentage of each FA in the TAG hydrolysate or its concentration in the studied matrix. The method exhibited high precision, as evidenced by the repeatability and within-lab reproducibility of results when tested on TAGs from both animal and vegetal origins. Compared to the analysis of FAMEs by GC-C-IRMS, the current procedure also brings several improvements in alignment with the principles of green analytical chemistry and green sample preparation. Thus, we present a two-in-one method for 13C-isotopomic and metabolomic biomarker quantitation within quasi-universal TAG compounds, encompassing the short- and medium-acyl chains.

Nice to know 2: The impact of NICE guidelines on ketogenic diet services in the UK and Ireland - An update.

Ketogenic diet therapy (KDT) has been recommended as a treatment for drug-resistant epilepsy in children and young people since 2012 in the National Institute for Health and Care Excellence Clinical Guidelines for Epilepsies. The Ketogenic Dietitians Research Network completed a survey in 2017 to assess the impact of these guidelines. An online survey was circulated to ketogenic dietitians across the UK and Ireland. The results were compared with those of the 2017 survey. The number of individuals following KDT was 854, comprising an increase of 13% since 2017. Service sizes ranged widely, with 1-74 (median 16) patients on the diet. Of 36 services, 30 had a waiting list, ranging from 2 to 67 (median 9) patients. The classical diet continued to be the most common KDT used (58% of patients). Ten services reported use of a new flexible medium chain triglyceride protocol. Some 48% of patients (n = 427) had been following the KDT for over 2 years, comprising an 18% increase since 2017. Of these, 68 (15.9%) had attempted to wean off KDT but had to re-start as a result of a deterioration in seizures. The number of individuals following medical KDT remains stable. Referral numbers and waiting lists remain high, highlighting that KDT is still a well-recognised treatment option for drug-resistant epilepsy. The types of KDT used are similar to previous years, although increasingly flexible protocols are being adopted. Longer-term use of KDT is increasing, with a proportion of patients requiring long-term use to maintain seizure control.

Sustainability Aspects of Carapa Guianensis Aubl Oil for Antiinflammatory Uses

Objective: The objective of this study is to describe the lipidomic profile of andiroba oil and its socio-economic characteristics focused on the activities of limonoids as an alternative for the generation of nonsteroidal anti- inflammatory drugs in order to relate the generation of products that act on circular and external peripheral inflammation. Theoretical Reference: In this context, andiroba oil (C. guianensis) has a wealth of lipid substances with chemical and socio-economic potential, highlighting its importance for local populations and its potential for sustainable bioeconomics, as this species contains nonsteroidal antiinflammatory drugs from Amazonian oil plants. Method: The methodology adopted includes describing the lipidomic profile of andiroba oil analyzed by GC/MS, using a Zebrom 5HT capillary column specifically designed to analyze both the methyl esters of the various fatty acids and the limonoids and steroids. The bibliographic search was carried out in the CAPES periodicals on the Scopus database, using specific keywords such as andiroba, Carapa, limonoids and antiinflammatories. Three samples of andiroba oil collected in different states were used to compare the lipidomic profiles. Results and Discussion: The results obtained revealed the presence of limonoids 7-desacetyl-7-oxogedunin, 7- oxogedunin, deacetyl gedunin; steroids campesterol, stigmasterol, b-sitosterol and various medium and long chain fatty acids. These results showed valuable information about the presence of limonoids, which have anti-inflammatory properties related to the chemical composition, which can vary with the season, the place of collection and the differences in the extraction processes of this oil. However, there is no total absence of limonoids. Research Implications: This research's practical and theoretical implications emphasize that the active anti-inflammatory principles are found in the unsaponifiable fraction, and therefore, only lipidomic techniques can access these metabolites with more excellent resolution. Thus, with the structural identifications, in silico studies can be carried out to classify their effects on cyclooxygenase enzymes (COXs), specifically COX-1, in circular and external peripheral inflammation. Originality/Value: This study contributes to the literature through the perspective of developing safe anti- inflammatory agents produced from andiroba oil, which is a source of great interest for topical use. This oil stands out as one of the most scientifically studied natural products, being biocompatible and presenting itself as an economical and safe alternative for treating circular and external peripheral inflammations. The relevance and value of this research is evidenced by the fact that limonoids are inhibitors of cyclooxygenases, highlighting the prospect of a new socio-economic aspect.

Lipidomics of sheep and goat Milk-based infant formulae during in vitro dynamic digestion

Lipid hydrolysis process during IF digestion, particularly the characterization of the lipidome and the resulting lipid breakdown products, has not been thoroughly investigated. This study aimed to compare the lipid hydrolysis profiles during the in vitro dynamic digestion of IFs made from whole sheep and goat milk. Using a lipidomics platform and multivariate statistical analysis, we observed changes in complex lipid levels during digestion. In the gastric compartment, we noted a progressive hydrolysis of triacylglycerols, phosphatidylcholines, and sphingomyelins. Conversely, lipolysis breakdown products like monoacylglycerols (e.g., MG(16:0), MG(18:0)), diacylglycerols, lysophosphatidylcholines (LPC 16:0, LPC 18:1, LPC 18:2), and free fatty acids increased in the intestinal compartment. The lipolysis trends were similar for both types of infant formulas, with long-chain fatty acid triglycerides (C > 46) exhibiting lower digestibility compared to medium-chain fatty acid triglycerides. Overall, these results indicate that sheep milk can be used as an ingredient in the manufacturing of IF.

Nutritional Management of Patients with Fatty Acid Oxidation Disorders.

Treatment of fatty acid oxidation disorders is based on dietary, pharmacological and metabolic decompensation measures. It is essential to provide the patient with sufficient glucose to prevent lipolysis and to avoid the use of fatty acids as fuel as far as possible. Dietary management consists of preventing periods of fasting and restricting fat intake by increasing carbohydrate intake, while maintaining an adequate and uninterrupted caloric intake. In long-chain deficits, long-chain triglyceride restriction should be 10% of total energy, with linoleic acid and linolenic acid intake of 3-4% and 0.5-1% (5/1-10/1 ratio), with medium-chain triglyceride supplementation at 10-25% of total energy (total MCT+LCT ratio = 20-35%). Trihepatnoin is a new therapeutic option with a good safety and efficacy profile. Patients at risk of rhabdomyolysis should ingest MCT or carbohydrates or a combination of both 20 min before exercise. In medium- and short-chain deficits, dietary modifications are not advised (except during exacerbations), with MCT contraindicated and slow sugars recommended 20 min before any significant physical exertion. Parents should be alerted to the need to increase the amount and frequency of carbohydrate intake in stressful situations. The main measure in emergency hospital treatment is the administration of IV glucose. The use of carnitine remains controversial and new therapeutic options are under investigation.

Chemometric characterization of the fatty acid and trace element profiles of organic and conventional Galician cheeses

The aim of this study was to evaluate the fatty acid profile and trace element content in three different Galician cow's milk cheeses (organic made from pasteurized milk, industrial made from pasteurized milk and artisanal made with raw milk). Twenty-five fatty acids and thirteen elements were determined by respectively gas liquid chromatography and inductively coupled plasma mass spectrometry, in thirty representative samples of the cheeses. The organic cheeses had significantly lower palmitic acid contents and significantly higher contents of odd and branched fatty acids, oleic acid, vaccenic acid, α-linolenic acid and conjugated linoleic acid than the other cheeses. The organic cheeses also had lower concentrations of trace and toxic element residues. Moreover, an association between medium chain fatty acids and palmitic acid and the presence of several mineral elements was detected by chemometric techniques. The nutritional evaluation of the organic cheeses was more favorable than for the other cheeses, as confirmed by the values of health-related indices.

Two-Stage Screening of Metschnikowia spp. Bioprotective Properties: From Grape Juice to Fermented Must by Saccharomyces cerevisiae.

Gluconobacter oxydans (Go) and Brettanomyces bruxellensis (Bb) are detrimental micro-organisms compromising wine quality through the production of acetic acid and undesirable aromas. Non-Saccharomyces yeasts, like Metschnikowia species, offer a bioprotective approach to control spoilage micro-organisms growth. Antagonist effects of forty-six Metschnikowia strains in a co-culture with Go or Bb in commercial grape juice were assessed. Three profiles were observed against Go: no effect, complete growth inhibition, and intermediate bioprotection. In contrast, Metschnikowia strains exhibited two profiles against Bb: no effect and moderate inhibition. These findings indicate a stronger antagonistic capacity against Go compared to Bb. Four promising Metschnikowia strains were selected and their bioprotective impact was investigated at lower temperatures in Chardonnay must. The antagonistic effect against Go was stronger at 16 °C compared to 20 °C, while no significant impact on Bb growth was observed. The bioprotection impact on Saccharomyces cerevisiae fermentation has been assessed. Metschnikowia strains' presence did not affect the fermentation time, but lowered the fermentation rate of S. cerevisiae. An analysis of central carbon metabolism and volatile organic compounds revealed a strain-dependent enhancement in the production of metabolites, including glycerol, acetate esters, medium-chain fatty acids, and ethyl esters. These findings suggest Metschnikowia species' potential for bioprotection in winemaking and wine quality through targeted strain selection.

Unlocking lager's flavour palette by metabolic engineering of Saccharomyces pastorianus for enhanced ethyl ester production

Despite being present in trace amounts, ethyl esters play a crucial role as flavour compounds in lager beer. In yeast, ethyl hexanoate, ethyl octanoate and ethyl decanoate, responsible for fruity and floral taste tones, are synthesized from the toxic medium chain acyl-CoA intermediates released by the fatty acid synthase complex during the fatty acid biosynthesis, as a protective mechanism. The aim of this study was to enhance the production of ethyl esters in the hybrid lager brewing yeast Saccharomyces pastorianus by improving the medium chain acyl-CoA precursor supply. Through CRISPR-Cas9-based genetic engineering, specific FAS1 and FAS2 genes harbouring mutations in domains of the fatty acid synthesis complex were overexpressed in a single and combinatorial approach. These mutations in the ScFAS genes led to specific overproduction of the respective ethyl esters: overexpression of ScFAS1I306A and ScFAS2G1250S significantly improved ethyl hexanoate production and ScFAS1R1834K boosted the ethyl octanoate production. Combinations of ScFAS1 mutant genes with ScFAS2G1250S greatly enhanced predictably the final ethyl ester concentrations in cultures grown on full malt wort, but also resulted in increased levels of free medium chain fatty acids causing alterations in flavour profiles. Finally, the elevated medium chain fatty acid pool was directed towards the ethyl esters by overexpressing the esterase ScEEB1. The genetically modified S. pastorianus strains were utilized in lager beer production, and the resulting beverage exhibited significantly altered flavour profiles, thereby greatly expanding the possibilities of the flavour palette of lager beers.

The beneficial impact of ketogenic diets on chemically-induced colitis in mice depends on the diet's lipid composition

Previously, we showed that restrictive diets, including ketogenic diet (KD), have an anti-inflammatory impact on the healthy gastrointestinal tract of mice. Afterward, we found that energy-restricting diets mitigate inflammation in the dextran sodium sulfate (DSS) colitis mouse model. The current study aimed to verify the impact of KD on DSS colitis and assess if the diet's fat composition influences the outcomes of the intervention.Mice with mild chronic colitis were fed control chow, KD composed of long-chain triglycerides (KD LCT) or a KD containing a mix of LCT and medium-chain triglycerides (KD LCT/MCT).KDs did not reverse DSS-enhanced gut permeability and shortening of the colon. Both KDs had a similar impact on liver, cecum, and spleen weight, villi and colon length, the thickness of muscularis externa, and expression of ZO-1 and occludin. On the contrary, body weight, glutathione (GSH) and taurine-GSH levels, GSH-S transferase (GST), and myeloperoxidase (MPO) activity, as well as an abundance of several fecal bacteria, all were differentially affected by the two types of KDs. When compared to the DSS control diet, reduction in colon mucosa cytokines expression was stronger in KD LCT than in the KD LCT/MCT group.We conclude that the outcomes of the KD interventions in terms of potential therapeutical applications depend on lipid composition. KD LCT showed a strong positive impact on gut inflammation. A potential contribution of GSH to KD outcomes and a correlation between MPO activity and microbiota composition was identified.

Continuous Liquid-Liquid Extraction of Medium-Chain Fatty Acids from Fermentation Broth Using Hollow-Fiber Membranes.

Medium-chain fatty acids (MCFAs; carbon-lengths: C6-C12) are high-value platform chemicals that serve a variety of industrial applications, including green antimicrobials, food ingredients, animal feed additives, cosmetics, fragrances, pharmaceuticals, and structured lipids. Currently, most MCFAs are produced from palm and coconut oil originating from Southeast Asia and South America. The conventional approach to harvesting palm and coconut fruits causes considerable ecological damage in these regions. Therefore, researchers are developing biological approaches (e.g., precision and open-culture fermentations) to generate MCFAs more sustainably using low-value substrates (e.g., methanol, ethanol, lactate) or organic wastes as feedstock. Microbial chain-elongation (CE) is a rapidly maturing open-culture fermentation platform that converts short-chain fatty acids (SCFAs; carbon lengths: C1-C5) into a subset of these MCFAs at industrially relevant rates. However, continuous in situ extraction of MCFA products is necessary not only to avoid product inhibition but also to facilitate the recovery of MCFAs in a pure and usable form.Liquid-liquid extraction (LLE) using hollow-fiber membranes and targeted extractant mixtures has proven a robust approach to selectively extract MCFA products from fermentation broths containing SCFAs. Here, the application of LLE for continuous MCFA removal is demonstrated using CE as the reference fermentation system and 3% (w/v) trioctylphosphine oxide in mineral oil as the extractant system. Fatty acids ranging from valeric acid (C5) to caprylic acid (C8) are selectively removed from SCFA-containing broths and concentrated to high titers in a semi-batch alkaline stripping solution for downstream processing.