Free Fatty Acid Profile Research Articles

Preparation of Pangasius hypophthalmus protein-stabilized pickering emulsions and 3D printing application

Pickering emulsions stabilized by protein have gotten many interests in recent years. The Basa fish (Pangasius hypophthalmus) is a popular aquatic product in the world, but its protein functions were seldom developed. This study reports food 3D printing inks constructed from Pickering emulsions by Basa fish protein stabilized and the profiles of its digestion in vitro. The excellent adsorption behavior of Basa fish protein (20 mg/mL) was applied to the interface of oil (60%) and water two-phase of Pickering emulsions. Rheological properties of crosslinking networks in the investigated Pickering emulsion might be adjusted to reach the required printability and extrudability during 3D printing. A slower release profile of free fatty acids was shown in the in vitro intestinal digestion experiment of Pickering emulsions. These edible Pickering emulsions have the potential to broaden 3D printing applications in foods and offer information about modulate the digestion of emulsified lipids with protein interface.

Substantial Fat Loss in Physique Competitors Is Characterized by Increased Levels of Bile Acids, Very-Long Chain Fatty Acids, and Oxylipins.

Weight loss and increased physical activity may promote beneficial modulation of the metabolome, but limited evidence exists about how very low-level weight loss affects the metabolome in previously non-obese active individuals. Following a weight loss period (21.1 ± 3.1 weeks) leading to substantial fat mass loss of 52% (−7.9 ± 1.5 kg) and low body fat (12.7 ± 4.1%), the liquid chromatography-mass spectrometry-based metabolic signature of 24 previously young, healthy, and normal weight female physique athletes was investigated. We observed uniform increases (FDR < 0.05) in bile acids, very-long-chain free fatty acids (FFA), and oxylipins, together with reductions in unsaturated FFAs after weight loss. These widespread changes, especially in the bile acid profile, were most strongly explained (FDR < 0.05) by changes in android (visceral) fat mass. The reported changes did not persist, as all of them were reversed after the subsequent voluntary weight regain period (18.4 ± 2.9 weeks) and were unchanged in non-dieting controls (n = 16). Overall, we suggest that the reported changes in FFA, bile acid, and oxylipin profiles reflect metabolic adaptation to very low levels of fat mass after prolonged periods of intense exercise and low-energy availability. However, the effects of the aforementioned metabolome subclass alteration on metabolic homeostasis remain controversial, and more studies are warranted to unravel the complex physiology and potentially associated health implications. In the end, our study reinforced the view that transient weight loss seems to have little to no long-lasting molecular and physiological effects.

The Quality and Composition of Iranian Low-Salt UF-White Cheese

In cheese, the reduction of salt is still a challenging task, as sodium chloride exerts multiple and fundamental functions. Salt favors the drainage of the residual whey; enhances the taste and the aroma profile; regulates the texture, the final pH, and the water activity; and affects the microbial growth. Hence the impact of partial replacement of NaCl by KCl on the ripening characteristics of Iranian UF (ultrafiltration) cheese during storage was monitored. To produce low-salt cheese, different mixtures of UF white cheese were treated with NaCl : KCl ratios of (a) 3% NaCl (control), (b) 1.50% NaCl+1.50% KCl, (c) 1.00% NaCl+2.00% KCl, and (d) 0.75% NaCl+2.25% KCl by dry salting. ADV (acid degree value) results showed significant differences ( P &lt; 0.05 ) in all treatments after 15, 30, 40, and 50 days of ripening. No significant differences were observed in the GC (gas chromatography) results in the samples’ free fatty acid (FFA) profile except for C18 : 0 in all treatments. KCl did not affect the moisture, dry matter, fat, TN (total nitrogen)/dry matter, and WSN (water-soluble nitrogen) contents of cheeses considerably. The evaluation of force to fracture showed that there were significant differences ( P &lt; 0.05 ) between treatment (d) as a control cheese and treatments (b) and (c). Sensory evaluations showed as the concentration of KCl increased, the cheese gradually became less acceptable and treatments with higher potassium chloride content were crumblier and less firm. Results of the aroma evaluation of cheese samples showed that unlike acetaldehyde, ethanol, acetoin, and diacetyl amounts decreased significantly ( P &lt; 0.05 ) during the storage period. Results also indicated that a reduction of sodium by up to 50% did not significantly affect the quality and composition of Iranian low-salt UF-white cheese except for sensory evaluation, texture analysis, and aroma characteristics.

Free fatty acid profiling of Greek yogurt by liquid chromatography-high resolution mass spectrometry (LC-HRMS) analysis

Yogurt is a fermented dairy product of high nutritional value, very popular in many parts of the world. Free fatty acids (FFAs), which are formed during fermentation, may cause changes in organoleptic properties of yogurt, and thus, the determination of FFAs is of importance. We present a liquid chromatography–high resolution mass spectrometry (LC-HRMS) method, which allows the simultaneous determination of a large set of common and uncommon FFAs in yogurt samples, avoiding any derivatization step. Twenty-five common saturated and unsaturated FAs, together with 21 saturated hydroxy fatty acids (SHFAs) and 17 saturated oxo fatty acids (SOFAs), were analyzed in 26 cow and 7 sheep Greek yogurt samples. A detailed analysis of bioactive SHFAs and SOFAs was carried out in yogurt samples for the first time. Differences at the concentrations of six common FAs and five oxidized FAs between the cow and sheep samples were observed. Based on these FAs, Principal Component Analysis (PCA) allows the discrimination of cow from sheep yogurt samples.

Reverting to a Healthy Diet during Lactation Normalizes Maternal Milk Lipid Content of Diet-Induced Obese Rats and Prevents Early Alterations in the Plasma Lipidome of the Offspring.

ScopeThis study aims to assess in rats whether normalizing maternal diet during lactation prevents the harmful effects of western diet (WD) consumption during the whole perinatal period on the lipidomic profile in maternal milk and offspring plasma.Methods and ResultsControl dams (CON‐dams), fed with standard diet (SD); WD‐dams, fed with WD prior and during gestation and lactation; and reversion dams (REV‐dams), fed as WD‐dams but moved to SD during lactation are followed. Lipidomic analysis is performed in milk and plasma samples from pups. Milk of WD‐dams presents a different triacylglycerol composition and free fatty acid (FA) profile compared to CON‐dams, including an increased ratio of pro‐inflammatory to anti‐inflammatory long‐chain polyunsaturated FA. Such alterations, which are also present in the plasma of their offspring, are widely reversed in the milk of REV‐dams and the plasma of their pups. This is related with the recovery of control adiponectin expression levels in the mammary gland, and the presence of decreased expression of pro‐inflammatory factors.ConclusionImplementing a healthy diet during lactation prevents early alterations in the plasma lipidome of pups associated to the maternal intake of an obesogenic diet, which may be related to the normalization of milk lipid content and the inflammatory state in the mammary gland.

Free fatty acids regulating action of Capparis decidua fruit on dyslipidemia in rats

Capparis decidua belongs to family Capparidaceae in wastelands of India. The study aim was to determine the role of C. decidua fruits on the free fatty acids (FFA) profile in fat-rich diet (FRD) dyslipidemic rats. The methanolic extract of edible fruit of C. decidua (CD) was given orally to obese dyslipidemic rats at the dose of 125 mg/kg and 250 mg/kg for consecutive 28 days. CD treatment in FRD rats significantly restricts the body weight gains. Blood lipid profile was altered dose dependently and significantly after 4-week treatment with CD to FRD. rats. It significantly (p&lt;0.05) enhanced serum FFA especially, g-linolinate, a-linolinate, arachidonate, ecosapentaenoate, docosapentaenoate and docosahexaenoate. Moreover, w3-PUFA content was also enhanced (50.3% and 78.8%) in the serum of CD treated animals, whereas MUFA was lowered (31.1% and 40%). Therefore, Capparis decidua fruit has a promising role on dyslipidemia and obesity and has the capabilities to regulate beneficial free fatty acids.

Effects of HIV Infection in Plasma Free Fatty Acid Profiles among People with Non-Alcoholic Fatty Liver Disease.

Despite its high prevalence, the mechanisms underlying non-alcoholic fatty liver disease (NAFLD) in people living with HIV (PLWH) are still unclear. In this prospective cohort study, we aim to evaluate differences in plasma fatty acid profiles between HIV-infected and HIV-uninfected participants with NAFLD. We included participants diagnosed with NAFLD, both HIV-infected and HIV-uninfected. Fatty acid methyl esters were measured from plasma samples. Ratios ([product]/[substrate]) were used to estimate desaturases and elongases activity. We used linear regression for adjusted analyses. We included 31 PLWH and 22 HIV-uninfected controls. We did not find differences in the sum of different types of FA or in FA with a greater presence of plasma. However, there were significant differences in the distribution of some FA, with higher concentrations of ALA, trans-palmitoleic, and behenic acids, and a lower concentration of lignoceric acid in PLWH. PLWH had lower C24:0/C22:0 and C16:0/C14:0 ratios, which estimates the activity of elongases ELOVL1 and ELOVL6. Both groups had similar fatty acid distribution, despite differences in traditional risk factors. PLWH had a lower proportion of specific ratios that estimate ELOVL1 and ELOVL6 activity, which had been previously described for other inflammatory conditions, such as psoriasis.

Follicular free fatty acid metabolic signatures and their effects on oocyte competence in non-obese PCOS patients.

Polycystic ovary syndrome (PCOS) is a common cause of anovulatory infertility in women. This study identified changes in free fatty acids profiles in the follicular fluid that may lead to better diagnosis and management of infertility in PCOS women. Polycystic ovary syndrome (PCOS) is a heterogeneous disease characterized by various endocrine/metabolic disorders and impaired reproductive potential. Alterations in oocyte competence are considered potentially causative factors for infertility in PCOS women and analyzing the composition of follicular fluid in these patients may help to identify which changes have the potential to alter oocyte quality. In this study, free fatty acid metabolic signatures in follicular fluid were performed to identify changes that may impact oocyte competence in non-obese PCOS women. Sixty-four non-obese women (32 with PCOS and 32 age- and BMI-matched controls) undergoing in vitro fertilization were recruited. Embryo quality was morphologically assessed. Free fatty acid metabolic profiling in follicular fluid was performed using gas/liquid chromatography-mass spectrometry. Principal component analysis and orthogonal partial least squares-discriminant analysis models were further constructed. Nine free fatty acids and 24 eicosanoids were identified and several eicosanoids synthesized by the cyclooxygenase pathway were significantly elevated in PCOS patients compared to controls. The combination of PGE2, PGF2α, PGJ2, and TXB2 had an area under the curve of 0.867 (0.775-0.960) for PCOS discrimination. Furthermore, follicular fluid levels of PGE2 and PGJ2 were negatively correlated with high-quality embryo rate in PCOS patients (P < 0.05). Metabolomic analysis revealed that follicular fluid lipidomic profiles undergo changes in non-obese PCOS women, which suggests that identifying changes in important metabolic signatures may give us a better understanding of the pathogenesis of PCOS. Furthermore, elevated PGE2 and PGJ2 concentrations may contribute to impaired oocyte competence in non-obese PCOS patients.

DOES FUNCTIONAL FOOD PROVIDE BENEFITS FOR CARDIOVASCULAR HEALTH? ONGOING STUDIES IN SCIENTIFIC CENTRE OF EXCELLENCE

Objective: Functional food (FF) is a food with presumably additional health benefits beyond its nutritional effects. We have been investigating health effects of consumption of hen eggs enriched with n-3 polyunsaturated fatty acids (n-3PUFAs) in healthy young sedentary populations, athletes and patients with acute and chronic coronary syndrome. Hereby, summarized data are presented to provide overall conclusions on the effect of n-3 PUFAs enriched eggs consumption on microvascular function, blood biochemical markers of inflammation and serum lipid profile of these participants. Design and method: Randomized controlled clinical interventional studies were conducted on a total of 90 healthy young (sedentary or athletes) and 40 cardiovascular participants. Participants were given n-3PUFAs enriched hen eggs (app.1 g n-3PUFAs (3 pcs)/ day for 3 consecutive weeks or regular eggs; n-3PUFAs 245 mg/day). Microvascular blood flow in response to various stimuli was assessed in skin microcirculation by laser Doppler flowmetry. Serum lipid profile, free fatty acids profile and various proinflammatory, anti-inflammatory and oxidative stress markers were measured. The study protocol and procedures conformed with the standards set by the latest revision of the Declaration of Helsinki, were approved by the Institutional Ethical Committees and registered at ClinicalTrials.gov. Results: Young healthy sedentaries on n-3 PUFAs eggs had significantly lower cholesterol and higher HDL-cholesterol after protocol. Microvascular flow was significantly increased after n-3 PUFAs consumption in sedentary and athletes. Glutathione peroxidase (GPx) and SOD activity decreased. Cardiovascular patients: LDL cholesterol and IL-6 (in Ac-CAD patients), and hsCRP and IL-1a (in all CAD patients) significantly decreased in n-3PUFAs group. GPx activity decreased, while microvascular reactivity remained unchanged. Conclusions: Consumption of FF can enhance microvascular function in healthy young participants and athletes; this effect is not observed in cardiovascular patients. The difference probably occurred due to the duration of the dietary protocol and medications taken by cardiovascular patients. Modest anti-inflammatory effects were observed in all study groups that consumed FF. Consumption of eggs does not have adverse effects on serum lipid profile. On the contrary; consumption of FF has a beneficial effect on the n-3 to n-6 free fatty acids ratio in serum of participants.

Effects of Orlistat or Telmisartan on the Serum Free Fatty Acids in Non-alcoholic Fatty Liver Disease Patients: An Open-Labeled Randomized Controlled Study.

One of the important inducers of inflammatory responses and accumulation of fat in hepatocytes is free fatty acids which ultimately lead to the development of non-alcoholic fatty liver disease. Patients with non-alcoholic fatty liver disease have high levels of plasma free fatty acids which are usually associated with type 2 diabetes and components of metabolic syndrome including dyslipidemia. Objective of this research is to investigate the effects of orlistat (a lipase enzyme inhibitor) or telmisartan (an angiotensin receptor blocker) on the serum free fatty acids in non-alcoholic fatty liver disease patients taking into consideration the baseline lipid profile. This open-label clinical trial was carried out in the Department of Pharmacology, College of Medicine at the University of Sulaimani in cooperation with Shar Teaching Hospital in Sulaimani city-Kurdistan Region of Iraq. A total number of 74 non-alcoholic fatty liver disease patients were recruited and grouped randomly into group I (n = 25) treated with orlistat (120 mg/day orally) for 12 weeks, group II (n = 24) treated with telmisartan (20 mg/day orally) for 8 weeks, and group III (n = 25) treated with placebo (carboxy- methyl cellulose) once daily. Fasting serum level of free fatty acid and lipid profile including total cholesterol, triglyceride, high-density lipoprotein, and non-high-density lipoproteins were determined. Orlistat and telmisartan significantly reduced the triglyceride-glucose index and free fatty acid levels (P < .001) in patients with non-alcoholic fatty liver diseases. Short-term treatment with orlistat or telmisartan produce effective and significant reductions in FFAs in patients with non-alcoholic fatty liver disease compared to placebo. Orlistat effectively reduces the free fatty acid irrespective of the baseline lipid profile.

Octanoic Acid-An Insecticidal Metabolite of Conidiobolus coronatus (Entomopthorales) That Affects Two Majors Antifungal Protection Systems in Galleria mellonella (Lepidoptera): Cuticular Lipids and Hemocytes.

The food flavour additive octanoic acid (C8:0) is also a metabolite of the entomopathogenic fungus Conidiobolus coronatus, which efficiently infects and rapidly kills Galleria mellonella. GC-MS analysis confirmed the presence of C8:0 in insecticidal fraction FR3 extracted from C. coronatus filtrate. Topical administration of C8:0 had a dose-dependent effect on survival rates of larvae but not on pupation or adult eclosion times of the survivors. Topically applied C8:0 was more toxic to adults than larvae (LD100 for adults 18.33 ± 2.49 vs. 33.56 ± 2.57 µg/mg of body mass for larvae). The administration of C8:0 on the cuticle of larvae and adults, in amounts corresponding to their LD50 and LD100 doses, had a considerable impact on the two main defense systems engaged in protecting against pathogens, causing serious changes in the developmental-stage-specific profiles of free fatty acids (FFAs) covering the cuticle of larvae and adults and damaging larval hemocytes. In vitro cultures of G. mellonella hemocytes, either directly treated with C8:0 or taken from C8:0 treated larvae, revealed deformation of hemocytes, disordered networking, late apoptosis, and necrosis, as well as caspase 1–9 activation and elevation of 8-OHdG level. C8:0 was also confirmed to have a cytotoxic effect on the SF-9 insect cell line, as determined by WST-1 and LDH tests.

Ginsenoside Rb1 Improves Metabolic Disorder in High-Fat Diet-Induced Obese Mice Associated With Modulation of Gut Microbiota.

Gut microbiota plays an important role in metabolic homeostasis. Previous studies demonstrated that ginsenoside Rb1 might improve obesity-induced metabolic disorders through regulating glucose and lipid metabolism in the liver and adipose tissues. Due to low bioavailability and enrichment in the intestinal tract of Rb1, we hypothesized that modulation of the gut microbiota might account for its pharmacological effects as well. Here, we show that oral administration of Rb1 significantly decreased serum LDL-c, TG, insulin, and insulin resistance index (HOMA-IR) in mice with a high-fat diet (HFD). Dynamic profiling of the gut microbiota showed that this metabolic improvement was accompanied by restoring of relative abundance of some key bacterial genera. In addition, the free fatty acids profiles in feces were significantly different between the HFD-fed mice with or without Rb1. The content of eight long-chain fatty acids (LCFAs) was significantly increased in mice with Rb1, which was positively correlated with the increase of Akkermansia and Parasuttereller, and negatively correlated with the decrease of Oscillibacter and Intestinimonas. Among these eight increased LCFAs, eicosapentaenoic acid (EPA), octadecenoic acids, and myristic acid were positively correlated with metabolic improvement. Furthermore, the colonic expression of the free fatty acid receptors 4 (Ffar4) gene was significantly upregulated after Rb1 treatment, in response to a notable increase of LCFA in feces. These findings suggested that Rb1 likely modulated the gut microbiota and intestinal free fatty acids profiles, which should be beneficial for the improvement of metabolic disorders in HFD-fed mice. This study provides a novel mechanism of Rb1 for the treatment of metabolic disorders induced by obesity, which may provide a therapeutic avenue for the development of new nutraceutical-based remedies for treating metabolic diseases, such as hyperlipidemia, insulin resistance, and type 2 diabetes.

Comparative study on the esterification of gymnodimine in different shellfish exposed to the dissolved toxin in seawater

Some lipophilic phycotoxins dissolved in seawater can be accumulated by bivalves via the filtering process. To explore the relationship between the bioaccumulation of gymnodimine-A (GYM-A) and free fatty acids (FFAs) of shellfish, three species of bivalves (venus clam Meretrix meretrix, mussel Mytilus galloprovincialis, and ark shell Anadara kagoshimensis) were exposed to dissolved GYM-A for 7 days in the same seawater system. Results indicated that GYM-A can be accumulated by these bivalves from the dissolved phase and esterified with FFAs reaching over 90% in most tissues of bivalves. Gymnodimine-A and its esters mainly distributed in the gills of shellfish, and the highest concentration of toxins occurred in mussel, followed by ark shell and venus clam. Similar percent of different fatty acid esters occurred in the experimental shellfish, in which the C16:0, C17:0, C18:0, C18:1, C20:1, C20:2, C22:2, and C22:6-GYM-A esters were the main metabolites of GYM-A. The binding capacity of fatty acids and GYM-A varied in different FFAs, which can explain why the C20:1-GYM-A ester dominated the ester profile while C16:0 was the most abundant fatty acid in all samples. Comparing with the FFA profile of shellfish in the control groups, overexpression of some FFAs occurred in the tissues of shellfish exposed to GYM-A in the experimental groups, which suggested that biosynthesis of FFAs was affected by the accumulation and metabolism of GYM-A in bivalves. Multiple fatty acids including some valuably nutritional FFAs such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were consumed in the esterification metabolism of GYM-A, which hinted that the lipid metabolism and nutritional quality of shellfish affected by the contamination of GYMs should be explored and assessed in future works.

Flazin as a Lipid Droplet Regulator against Lipid Disorders.

Lipid disorders are closely related to numerous metabolic diseases, and lipid droplets (LDs) have been considered as a new target for regulating lipid metabolism. Dietary intervention and nutraceuticals provide safe and long-term beneficial effects for treating metabolic diseases. Flazin is a diet-derived bioactive constituent mainly existing in fermented foods, of which the lipid metabolism improvement function has not been studied. In this study, the effect of flazin on lipid regulation at both cell level and organelle level was investigated. Lipidomic profiling showed that flazin significantly decreased cellular triglyceride (TG) by 12.0–22.4% compared with modeling groups and improved the TG and free fatty acid profile. LD staining revealed that flazin efficiently reduced both cellular neutral lipid content by 17.4–53.9% and LD size by 10.0–35.3%. Furthermore, nanoelectrospray ionization mass spectrometry analysis proved that flazin exhibited a preferential suppression of LD TG and regulated LD morphology, including a size decrease and surface property improvement. An evaluation of related gene expression suggested the mechanism to be lipolysis promotion and lipogenesis inhibition. These findings indicated that flazin might be an LD regulator for reversing lipid metabolism disturbance. Moreover, the strategy proposed in this study may contribute to developing other nutraceuticals for treating lipid disorder-related metabolic diseases.

The combination of RPLC-ESI-FTMS/MS and m-CPBA epoxidation for the location and geometry assignment of double bonds in unsaturated fatty acyl chains of drying oils

Abstract Oil painting is the process of painting with drying oils as binders. Such oils mainly consist of triacylglycerols characterized by high levels of unsaturated fatty acyl chains. For instance, poppy seed oils have received much attention for their use as oil paints since very ancient time. Upon exposure to oxygen, not only do drying oils harden to a densely cross-linked polymer network through radical reactions, but their fatty acids also interact with cations of some pigments forming the related carboxylate salts. These so-called metal soaps lead to severe degradation phenomena because they can migrate and aggregate, breaking through the surface of the paint. Since the fatty-acidome of a drying oil is involved in these processes, the current study is chiefly devoted to the identification of both free and total fatty acids in poppy seed oil by reversed-phase liquid chromatography coupled with high-resolution mass spectrometry by an electrospray ionization source (RPLC-ESI-FTMS). Total fatty acids (TFA) were investigated by enzymatic hydrolysis and comparisons were made between the free fatty acid (FFA) profile and the total one resulting from the digestion on the same poppy seed oil. A key component of our approach was the double bond (C=C) epoxidation reaction, which was carried out on unsaturated fatty acids by meta-chloroperoxybenzoic acid (m-CPBA). A chromatographic set up was developed to separate such oxidized lipids, that were identified as deprotonated molecules [epoM-H]− by accurate mass matching in full mass spectra. Their epoxide ring was subsequently cleaved upon tandem mass fragmentation to yield a pair of diagnostic product ions for C=C double bond localization. Interestingly, the double bond geometry (i.e., cis/trans) was also assigned based on the retention times of derivatized-FA. The present approach opens new avenues for the analysis of artworks whether affected by metal soap-related degradation phenomena.

STUDY ON THE UTILIZATION OF CATECHINS FROM GAMBIR (UNCARIA GAMBIR ROXB) LEAVES AS ANTIOXIDANTS COOKING OIL

Catechins are flavonoid molecules that act as antioxidants in the body. The degradation of palm oil with catechin treatment (400, 200, 100, 50, 0) was investigated in this work (ppm). The goal of this study was to see how efficient catechins are as antioxidants in cooking oil and what the optimal treatment would be for the amount of catechins added. Catechin concentration, antioxidant IC50, free fatty acid, peroxide value, water content, color, and fatty acid profile are all analyzed using GC-MS. Treatment E (400 ppm) was found to be the most effective, with free fatty acid values of 0.45 percent, peroxide value of 5 mek/g, moisture content of 0.89 percent, and color of 95.95 (L*16, 11, a* -1.51, b* 14.51). Catechins can retain three fatty acids in the oil, according to GC-MS analysis: hexadecanoic acid, 9-octadecanoic acid, and heptadecanoic acid.

Replacement of Pregastric Lipases in Cheese Production: Identification and Heterologous Expression of a Lipase from Pleurotus citrinopileatus.

Traditionally produced piquant cheeses such as Feta or Provolone rely on pregastric lipolytic enzymes of animal origin to intensify flavor formation during ripening. Herein, we report a novel fungal lipase, derived from the phylum Basidiomycota to replace animal-derived products. A screening of 31 strains for the desired hydrolytic activities was performed, which revealed a promising fungal species. The secretome of an edible golden oyster mushroom, Pleurotus citrinopileatus, provided suitable enzymatic activity, and the coding sequence of the corresponding enzyme was identified by combining transcriptome and liquid chromatography high-resolution electrospray tandem mass spectroscopy (LC-HR-ESI-MS/MS) data. Recombinant expression in Escherichia coli BL21 (DE3) using chaperones GroES-GroEL and DnaK-DnaJ-GrpE was established. The recombinant lipolytic enzyme was purified and biochemically characterized in terms of thermal and pH stability, optimal reaction conditions, and kinetic data toward p-nitrophenyl esters. An application in the microscale production of Feta-type brine cheese revealed promising sensory properties, which were confirmed by headspace solid-phase microextraction gas chromatography mass spectrometry (HS-SPME-GC-MS) analyses in comparison with the reference enzyme opti-zym z10uc from goat origin. Supplementation with 2.3 U of the heterologously expressed fungal lipase produced the most comparable free fatty acid profile after 30 days of ripening. The flavor and texture formed during the application of the new lipase from P. citrinopileatus proved to be competitive to the use of pregastric lipases and could therefore replace the products of animal origin.

Serum Levels of Free Fatty Acids in Obese Mice and Their Associations with Routine Lipid Profiles.

PurposeTo investigate serum levels of free fatty acids (FFAs) and their associations with routine serum lipids in diet-induced obese mice, which have been scantily reported before.MethodsMale C57BL/6 J mice were fed high-fat diets for 12 weeks to induce obesity. Levels of serum FFAs were measured by ultra-performance liquid chromatography-electrospray ionization-tandem mass spectrometry.ResultsObese mice had higher serum levels of total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C), but lower triglycerides (TG) than control mice. A total of 30 FFAs were found, and 3 saturated fatty acids (SFAs), all 8 monounsaturated fatty acids (MUFAs) and 7 polyunsaturated fatty acids (PUFAs) decreased in obese mice, but one SFA (C4:0) increased. Differences in the relative levels of individual FFAs to total FFAs, SFAs, MUFAs or PUFAs between obese and control mice were different from each other and from those evaluated by concrete levels except C4:0, C16:1, C19:1 and C18:4. Only the concrete levels of C4:0, C22:3 and C18:4 were associated with routine serum lipids, including C22:3 negatively with TG in control mice, and C4:0 and C18:4 positively with LDL-C in obese mice, although the relative levels of C4:0 to total MUFAs negatively with TC, and C23:3 to total SFAs or MUFAs negatively with TG in control mice. Different relative levels of the remaining FFAs were differently associated with different routine serum lipids in obese and/or control mice.ConclusionObesity may influence serum FFAs profiles. The relationship of individual FFAs and their relative levels to other FFAs with routine serum lipids in obese and control mice suggests that individual FFAs may interact with others and obesity on levels of routine serum lipids. Once confirmed, the interactions may be novel perspectives when fatty acids are used to improve hyperlipidemia in the subjects with obesity.

Estimated Elovl6 and delta-5 desaturase activities might represent potential markers for insulin resistance in Japanese adults.

Results from a recent study indicated that lower stearic acid/palmitic acid (SA/PA) and arachidonic acid/dihomo-γ-linolenic acid (AA/DGLA) ratios were associated with metabolically unhealthy obesity. However, this has not been extensively studied in the Japanese population. We recruited 291 Japanese subjects with serum free fatty acid profiles undergoing health examinations. Whole serum desaturase activity was estimated as the product: precursor ratio -SA/PA ratio for elongation of long-chain fatty acid family member 6 (Elovl6) and AA/DGLA for delta-5 desaturase (D5D). The determinants of Elovl6 and D5D activity were investigated using multiple regression analyses. The Elovl6 and D5D activities exhibited a negative correlation with the logmatic-transformed TG/HDL-C ratio and TyG index. Multiple regression analyses revealed that the TG/HDL-C ratio and TyG index were negatively associated with Elovl6 and D5D activities. Most atherogenic markers were worse in the low Elovl6 or D5D activity group than in the high Elovl6 or D5D activity group. When study subjects were further stratified by TG levels, most atherogenic markers were the worst in the highest TG group in either the lowest Elovl6 or lowest D5D activity groups. The estimated Elovl6 and D5D activities might be useful markers of insulin resistance in Japanese subjects.

A phospholipase B from Pseudomonas aeruginosa with activity towards endogenous phospholipids affects biofilm assembly

Pseudomonas aeruginosa is a severe threat to immunocompromised patients due to its numerous virulence factors and biofilm-mediated multidrug resistance. It produces and secretes various toxins with hydrolytic activities including phospholipases. However, the function of intracellular phospholipases for bacterial virulence has still not been established. Here, we demonstrate that the hypothetical gene pa2927 of P. aeruginosa encodes a novel phospholipase B named PaPlaB. At reaction equilibrium, PaPlaB purified from detergent-solubilized membranes of E. coli released fatty acids (FAs) from sn-1 and sn-2 positions of phospholipids at the molar ratio of 51:49. PaPlaB in vitro hydrolyzed P. aeruginosa phospholipids reconstituted in detergent micelles and phospholipids reconstituted in vesicles. Cellular localization studies indicate that PaPlaB is a cell-bound PLA of P. aeruginosa and that it is peripherally bound to both membranes in E. coli, yet the active form was predominantly associated with the cytoplasmic membrane of E. coli. Decreasing the concentration of purified and detergent-stabilized PaPlaB leads to increased enzymatic activity, and at the same time triggers oligomer dissociation. We showed that the free FA profile, biofilm amount and architecture of the wild type and ΔplaB differ. However, it remains to be established how the PLB activity of PaPlaB is regulated by homooligomerisation and how it relates to the phenotype of the P. aeruginosa ΔplaB. This novel putative virulence factor contributes to our understanding of phospholipid degrading enzymes and might provide a target for new therapeutics against P. aeruginosa biofilms.