Hydrolysis Of Triglycerides Research Articles

Abstract 121: Exploiting Natural Genetic Variation In The Human Triglyceride Regulator APOA5 To Understand Its Function

Plasma triglycerides (TGs) are an independent predictor of the risk for CAD, the leading cause of death worldwide. TGs are also positively associated with risk and severity of hyperTG-induced acute pancreatitis (HTG-AP). Current therapies are often insufficient in reducing extremely elevated TGs. We believe that apolipoprotein A-V (apoA-V, encoded by APOA5) can fill this unmet medical need. ApoA-V is a potent modulator of TG metabolism; it enhances lipoprotein lipase TG hydrolysis. We hypothesize that naturally occurring human APOA5 variants can inform ApoA-V function and identify novel ApoA-V based therapeutic axes. We used the Penn Medicine Biobank (PMBB) to identify and measure plasma TGs of carriers of APOA5 variants predicted to change ApoA-V structure-function. Then, we used hydrogen-deuterium exchange mass spectroscopy to determine the secondary structure of ApoA-V, thereby identifying putative functional domains onto which we mapped our variants of interest. Finally, we characterized the plasma lipid effects of these mutants using adeno-associated viral (AAV) vectors in apoa5 knockout (KO) mice. We identified APOA5 variants associated with changes in plasma TGs. These variants primarily fall near the central heparin binding domain or C-terminal lipid binding domain. We selected APOA5 Q275X, which removes the entire lipid binding domain, for further interrogation. Apoa5 KO mice that received APOA5 Q275X AAV had higher plasma TGs than mice treated with WT APOA5 AAV. While WT ApoA-V protein associated with VLDL and HDL particles, Q275X ApoA-V protein appeared in lipoprotein free fractions. We have identified APOA5 variants associated with plasma TG phenotypes in humans, and mapped them to an experimentally determined ApoA-V secondary structure to identify the functional domains likely impacted. We have identified APOA5 Q275X as a loss of function variant that fails to bind lipoprotein particles and is associated with elevated plasma TGs. Continued study of this and other interesting naturally occurring variants will provide insight into the function of ApoA-V in TG metabolism. These insights can help us to therapeutically enhance ApoA-V to rapidly reduce TG levels during acute HTG-AP and to help prevent recurrent HTG-AP.

Understanding Metabolic Remodeling in Mycobacterium smegmatis to Overcome Energy Exigency and Reductive Stress Under Energy-Compromised State.

Mycobacteria such as Mycobacterium tuberculosis, the causative agent of tuberculosis that annually kills several million people worldwide, and Mycobacterium smegmatis, the non-pathogenic fast-growing mycobacteria, require oxidative phosphorylation to meet their energy requirements. We have previously shown that deletion of one of the two copies of atpD gene that codes for the ATP synthase β-subunit establishes an energy-compromised state in M. smegmatis. Here we report that upon such deletion, a major routing of electron flux occurs through the less energy-efficient complexes of its respiratory chain. ΔatpD bacterium also shows an increased reduced state which is further confirmed by the overexpression of WhiB3, a major redox sensor. We show a substantial modulation of the biosynthesis of cell wall associated lipids and triacylglycerol (TAG). An accumulation of TAG-containing lipid bodies is further confirmed by using 14C oleate incorporation. Interestingly, the mutant also shows an overexpression of TAG-degrading lipase genes, and the intracellular lipolytic enzymes mediate TAG hydrolysis for their utilization as energy source. We believe that our in vitro energy-depleted model will allow us to explore the critical link between energy metabolism, redox homeostasis, and lipid biosynthesis during ATP-depleted state, which will enhance our understanding of the bacterial adaptation, and will allow us to identify novel drug targets to counter mycobacterial infections.

Association between the levels of CGI-58 and lipoprotein lipase in the placenta of patients with preeclampsia.

Preeclampsia is an idiopathic disease of pregnancy, which seriously endangers the life of both the mother and the infant. The pathogenesis of preeclampsia has not been fully elucidated, although it is generally considered to be associated with abnormal lipid metabolism during pregnancy. Comparative gene identification-58 (CGI-58) and lipoprotein lipase (LPL) are involved in the first step of triglyceride hydrolysis and serve an important role in lipid transport in the placenta. The present study aimed therefore to investigate the association between CGI-58 and LPL in the placentas of patients with or without preeclampsia and to evaluate blood lipid levels. The patient cohort was divided into two groups, pregnant women with preeclampsia and normal pregnant women (control). According to biochemical analyses, reverse transcription-quantitative PCR, immunohistochemistry analysis and western blotting, the expression of CGI-58 and LPL in the placenta was detected, the blood lipid levels were evaluated and other clinical data were collected. Compared with the control group, triglycerides (TGs), low density lipoprotein-cholesterol (LDL-C), apolipoprotein B (ApoB) and atherosclerotic index (AI) were significantly higher in the preeclampsia group, whereas high density lipoprotein-cholesterol (HDL-C) and apolipoprotein A (ApoA) were significantly lower (P<0.05). Furthermore, the expression levels of CGI-58 and LPL in the placental tissue of the preeclampsia group was significantly lower than that of the control group (P<0.05). Linear correlation analysis demonstrated that there was a positive association between CGI-58 and LPL (r=0.602; P<0.05), that CGI-58 was positively associated with HDL-C (r=0.63; P<0.01) but negatively associated with TG and ApoB (r=0.840; P<0.01; and r=0.514; P<0.05, respectively), that LPL was positively associated with HDL-C (r=0.524; P<0.01) but negatively associated with TG and AI (r=0.659; P<0.01; and r=0.496; P<0.01, respectively). These results suggested that the expression of CGI-58 and LPL in the placenta was associated with the pathogenesis of preeclampsia and maternal lipids and the risk of preeclampsia was increased with decreasing expression levels of CGI-58 and LPL. Hence, CGI-58 and LPL may be used as important indicators for the diagnosis of preeclampsia and for the prevention of preeclampsia in pregnant women.

Thioesterase induction by 2,3,7,8-tetrachlorodibenzo-p-dioxin results in a futile cycle that inhibits hepatic \u03b2-oxidation

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD), a persistent environmental contaminant, induces steatosis by increasing hepatic uptake of dietary and mobilized peripheral fats, inhibiting lipoprotein export, and repressing β-oxidation. In this study, the mechanism of β-oxidation inhibition was investigated by testing the hypothesis that TCDD dose-dependently repressed straight-chain fatty acid oxidation gene expression in mice following oral gavage every 4 days for 28 days. Untargeted metabolomic analysis revealed a dose-dependent decrease in hepatic acyl-CoA levels, while octenoyl-CoA and dicarboxylic acid levels increased. TCDD also dose-dependently repressed the hepatic gene expression associated with triacylglycerol and cholesterol ester hydrolysis, fatty acid binding proteins, fatty acid activation, and 3-ketoacyl-CoA thiolysis while inducing acyl-CoA hydrolysis. Moreover, octenoyl-CoA blocked the hydration of crotonyl-CoA suggesting short chain enoyl-CoA hydratase (ECHS1) activity was inhibited. Collectively, the integration of metabolomics and RNA-seq data suggested TCDD induced a futile cycle of fatty acid activation and acyl-CoA hydrolysis resulting in incomplete β-oxidation, and the accumulation octenoyl-CoA levels that inhibited the activity of short chain enoyl-CoA hydratase (ECHS1).

Morphine increases myocardial triacylglycerol through regulating adipose triglyceride lipase S406 phosphorylation

AimsMorphine, a commonly used drug for anesthesia, affects lipid metabolism in different tissues, but the mechanism is currently unclear. Adipose triglyceride lipase (ATGL) is the rate-limiting enzyme responsible for the first step of triglyceride (TG) hydrolysis. Here we aim to investigate whether ATGL phosphorylation is involved in morphine-induced TG accumulation. Main methodsOil red O staining and TG content analysis were used to detect the effect of morphine on lipid storage. A series of ATGL phosphoamino acid site mutant plasmids were constructed by gene synthesis and transfected to HL-1 cells to evaluate the phosphorylation levels of ATGL phosphoamino acid in morphine-treated HL-1 cells with immunoprecipitation and immunoblotting assay. Key findingsMorphine acute treatment induced excessive accumulation of TG and decreased the phosphorylation level of ATGL Ser406 in HL-1 cells. Of note, the phosphorylation positive mutation of ATGL Ser406 to aspartic acid effectively reversed morphine-induced excessive accumulation of TG in HL-1 cells. SignificanceThis discovery will help to fully understand the lipid regulation function of morphine in a new scope. In addition, it will expand the phosphorylation research of ATGL more comprehensively and provide powerful clues for lipid metabolism regulation.

Lysosomal acid lipase controls dendritic cells maturation

Background and Aims: Lysosomal acid lipase (LAL) hydrolyzes cholesterol esters and triglycerides thus releasing free cholesterol and fatty acids. In immune cells, these metabolites contribute to cellular metabolic reprogramming and indeed, LAL was shown to be critical for the polarization of macrophages toward pro-resolutive phenotype and the generation of memory T cell pools. Aim of this project is to investigate the role of LAL during dendritic cells development and maturation.

Hypertriglyceridemia and acute pancreatitis complicated by splenic and hepatic infarcts

Background and Aims: Hypertriglyceridemia is a common clinical problem that can be exacerbated by numerous medications and medical conditions. Primary hypertriglyceridemia caused by lipoprotein lipase (LPL) deficiency is an extremely rare metabolic disorder. LPL is a lipolytic enzyme involved in catalyzing hydrolysis of triglycerides (TG) and very low-density lipoprotein (VLDL) particles.

Regulation of lipid homeostasis by the TBC protein dTBC1D22 via modulation of the small GTPase Rab40 to facilitate lipophagy.

The regulation of lipid homeostasis is not well understood. Using forward genetic screening, we demonstrate that the loss of dTBC1D22, an essential gene that encodes a Tre2-Bub2-Cdc16 (TBC) domain-containing protein, results in lipid droplet accumulation in multiple tissues. We observe that dTBC1D22 interacts with Rab40 and exhibits GTPase activating protein (GAP) activity. Overexpression of either the GTP- or GDP-binding-mimic form of Rab40 results in lipid droplet accumulation. We observe that Rab40 mutant flies are defective in lipid mobilization. The lipid depletion induced by overexpression of Brummer, a triglyceride lipase, is dependent on Rab40. Rab40 mutant flies exhibit decreased lipophagy and small size of autolysosomal structures, which may be due to the defective Golgi functions. Finally, we demonstrate that Rab40 physically interacts with Lamp1, and Rab40 is required for the distribution of Lamp1 during starvation. We propose that dTBC1D22 functions as a GAP for Rab40 to regulate lipophagy.

AAV5-based gene therapy targeting ANGPTL3 reduces hyperlipidemia and development of atherosclerosis in APOE*3-Leiden.CETP mice

Background and Aims: ANGPTL3 is an inhibitor of lipoprotein and endothelial lipase, thus restricting triglyceride hydrolysis from triglyceride-rich lipoproteins and leading to increased plasma lipid levels. We explored the therapeutic potential of a gene therapeutic approach to target ANGPTL3 expression by introduction of an artificial microRNA into hepatocytes using an adeno-associated virus type 5 (AAV5) vector (AAV5-miQURE™) in hyperlipidemic APOE*3-Leiden.CETP mice, alone or in combination with atorvastatin treatment.

Engineering of Saccharomyces cerevisiae for the accumulation of high amounts of triacylglycerol

BackgroundFatty acid-based substances play an important role in many products, from food supplements to pharmaceutical products and biofuels. The production of fatty acids, mainly in their esterified form as triacylglycerol (TAG), has been intensively studied in oleaginous yeasts, whereas much less effort has been invested into non-oleaginous species. In the present work, we engineered the model yeast Saccharomyces cerevisiae, which is commonly regarded as non-oleaginous, for the storage of high amounts of TAG, comparable to the contents achieved in oleaginous yeasts.ResultsWe investigated the effects of several mutations with regard to increased TAG accumulation and identified six of them as important for this phenotype: a point mutation in the acetyl-CoA carboxylase Acc1p, overexpression of the diacylglycerol acyltransferase Dga1p, deletions of genes coding for enzymes involved in the competing pathways glycogen and steryl ester synthesis and TAG hydrolysis, and a deletion of CKB1, the gene coding for one of the regulatory subunits of casein kinase 2. With the combination of these mutations in a S. cerevisiae strain with a relatively high neutral lipid level already in the non-engineered state, we achieved a TAG content of 65% in the dry biomass. High TAG levels were not only obtained under conditions that favor lipid accumulation, but also in defined standard carbon-limited media.ConclusionsBaker's yeast, which is usually regarded as inefficient in the storage of TAG, can be converted into a highly oleaginous strain that could be useful in processes aiming at the synthesis of fatty acid-based products. This work emphasizes the importance of strain selection in combination with metabolic engineering to obtain high product levels.

Disturbance of lipid homeostasis in lysosomal lipase deficiency – pathomechanism, diagnosis and treatment

Lysosomal acid lipase (LAL) plays a key role in lipid metabolism through the hydrolysis of cholesteryl esters and triglycerides in lysosomes. LAL deficiency is a rare autosomal recessive lysosomal storage disease caused by deleterious mutations in the LIPA gene. In the case of LAL deficiency, cholesteryl esters and triglycerides accumulate within the lysosomes. The up-regulation of endogenous cholesterol production, increased synthesis of apolipoprotein B (ApoB) and increased production of very-low-density lipoprotein cholesterol (VLDL-C) is observed. The diagnosis is easy due to the currently available method of testing the enzyme activity in a dry blood spot. Molecular analysis is necessary to verify the clinical and biochemical diagnosis and to analyze the genotype-phenotype correlation. Sebelipase alfa is a recombinant human lysosomal lipase intended for use in enzyme replacement therapy in patients with LAL deficiency.

The disassembly of lipid droplets in Chlamydomonas.

Lipid droplets (LDs) are ubiquitous and specialized organelles in eukaryotic cells. Consisting of a triacylglycerol core surrounded by a monolayer of membrane lipids, LDs are decorated with proteins and have myriad functions, from carbon/energy storage to membrane lipid remodeling and signal transduction. The biogenesis and turnover of LDs are therefore tightly coordinated with cellular metabolic needs in a fluctuating environment. Lipid droplet turnover requires remodeling of the protein coat, lipolysis, autophagy and fatty acid β-oxidation. Several key components of these processes have been identified in Chlamydomonas (Chlamydomonas reinhardtii), including the major lipid droplet protein, a CXC-domain containing regulatory protein, the phosphatidylethanolamine-binding DTH1 (DELAYED IN TAG HYDROLYSIS1), two lipases and two enzymes involved in fatty acid β-oxidation. Here, we review LD turnover and discuss its physiological significance in Chlamydomonas, a major model green microalga in research on algal oil.

Eruptive xanthoma model reveals endothelial cells internalize and metabolize chylomicrons, leading to extravascular triglyceride accumulation.

Although tissue uptake of fatty acids from chylomicrons is primarily via lipoprotein lipase (LpL) hydrolysis of triglycerides (TGs), studies of patients with genetic LpL deficiency suggest additional pathways deliver dietary lipids to tissues. Despite an intact endothelial cell (EC) barrier, hyperchylomicronemic patients accumulate chylomicron-derived lipids within skin macrophages, leading to the clinical finding eruptive xanthomas. We explored whether an LpL-independent pathway exists for transfer of circulating lipids across the EC barrier. We found that LpL-deficient mice had a marked increase in aortic EC lipid droplets before and after a fat gavage. Cultured ECs internalized chylomicrons, which were hydrolyzed within lysosomes. The products of this hydrolysis fueled lipid droplet biogenesis in ECs and triggered lipid accumulation in cocultured macrophages. EC chylomicron uptake was inhibited by competition with HDL and knockdown of the scavenger receptor-BI (SR-BI). In vivo, SR-BI knockdown reduced TG accumulation in aortic ECs and skin macrophages of LpL-deficient mice. Thus, ECs internalize chylomicrons, metabolize them in lysosomes, and either store or release their lipids. This latter process may allow accumulation of TGs within skin macrophages and illustrates a pathway that might be responsible for creation of eruptive xanthomas.

Chronic Elevation of Tumor Necrosis Factor-Alpha Suppresses Lactation via Downregulation of Lipoprotein Lipase

ObjectivesIdentification of physiologic factors that impede lactation is critical for developing interventions to support milk production. Our objective was to examine the role of tumor necrosis factor–alpha (TNF-A), a potent regulator of lipid metabolism, in the suppression of human lactation. We hypothesized that mothers with very low, versus sufficient milk production, would have elevated milk TNF-A and differential expression of TNF-A signaling pathways in the mammary epithelial cells (MECs). MethodsAmong mothers in a study of low milk supply, extracellular mammary epithelial cell mRNA was isolated from fresh milk fat and submitted to RNA-sequencing. Aligned and quantified reads were examined for differentially expressed genes using t-tests (DEG, p < 0.05 after false discovery rate adjustment). We compared DEG in n = 5 with the lowest [LMP] (0–53 mL/24 h), and n = 4 with the highest [HMP] (422–463 mL/24 h) milk production in the sampled breast, using PPAR, TNF-Alpha, and fatty acid biosynthesis KEGG pathways. BMI, fasting serum triglycerides, and milk TNF-A were also assessed. ResultsMean day postpartum (StdDev) was 32 (11) versus 47 (19) in LMP versus HMP, P > 0.05. Serum fasting triglyceride and BMI were significantly elevated in LMP versus HMP, P < 0.05. Median [min-max] TNF-A was 1.77 [0.64–6.89] pg/mL in LMP versus none detected in HMP (limit of detection, 0.50 pg/mL). Transcriptome analysis revealed 1.7-fold down regulation of lipoprotein lipase (LPL) in LMP versus HMP. PPARA and RXRA, regulatory factors upstream of LPL, were downregulated 1.8-fold. There was no difference in the expression of genes directly involved in fatty acid biosynthesis. ConclusionsMothers with insufficient milk production have elevated milk TNF-A and a corresponding reduction in expression of lipid processing proteins known to be downregulated by TNF-A, notably LPL which is necessary for the hydrolysis of triglycerides into free fatty acids and glycerol for transport into MECs. We propose that TNF-A’s disruption of critical fatty acid transport deprives the mammary gland of vital fuel and substrate towards the synthesis of human milk, without compensation through de novo lipid synthesis. Our findings shed light on TNF-A mediated lipid dysregulation as a potential link between obesity and insufficient lactation. Funding SourcesNIH and USDA.

197-LB: The Regulation of HSL by Macrophage Migration Inhibitory Factor (MIF) Contributes to Adipocyte Hypertrophy and Development of Obesity

Inflammation is associated with lipolysis in adipose tissue. HSL is a key enzyme in mediating lipolysis through augmenting hydrolysis of triglycerides and diacylglycerides. Our study found that the procytokine MIF has both transcriptional and non-transcriptional effects on mediating HSL through activating AMPK signaling pathway. The activated AMPK downregulates HSL gene expression in adipocytes. It also upregulates a phosphorylation of HSL Ser565, an inhibitory phosphorylation site, leading to the reduction of HSL active site, Ser563 phosphorylation. These mechanisms are operative in either transgenic overexpressed MIF mouse models or WT mice with high circulating MIF levels following a high caloric diet feeding (HFD). The MIF/AMPK/HSL axis regulates the development of adipocyte hypertrophy and obesity. We also confirm that high plasma MIF levels are associated with decreased adipose HSL gene expression in human obesity. Interestingly, in Mif-/-mice, HSL gene expression is also inhibited, suggesting that MIF genotype is important to maintain adipose HSL expression. Following HFD feeding, Mif-/-mice demonstrates higher Akt and lower PKA phosphorylation, which further decrease HSL activation and exacerbate obesity compared to WT. Thus, HSL regulation by MIF may be an important molecular mechanism in the development of obesity.View largeDownload slideView largeDownload slide DisclosureL. Chen: None. Y. Huang: None. L. Li: None. H. Zabihi: None. Y. Qi: None. L. Leng: None. G. Sun: None. R. Bucala: None. D. Qi: None.FundingNational Sciences and Engineering Research Council of Canada (RGPIN-2017-04542); Canadian Institutes of Health Research (PJT-156116)

The influence of RS738409 I148M polymorphism of patatin-like phospholipase domain containing 3 gene on the susceptibility of non-alcoholic fatty liver disease

We aimed to elucidate the frequency of polymorphic genotypes and alleles of patatin-like phospholipase domain containing 3 rs738409 polymorphism and its possible associations with non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis in a cohort from Turkey.We enrolled 200 patients diagnosed with NAFLD and genotyped for rs738409 I148M polymorphism by real-time polymerase chain reaction, particularly by melting curve analysis. SPSS analysis software was used for statistical significance. Continuous variable values were expressed as mean ± standard deviation. Significant statistical level was chosen as p = 0.05.Our results demonstrate in a cohort from Turkey that rs738409 C > G polymorphism (I148M) of patatin-like phospholipase domain containing 3 gene is significantly able to affect individuals to have NAFLD in unadjusted regression model.Consistent with the previous studies in other populations, our study group showed a significantly higher risk of having NAFLD in unadjusted regression model but not in the adjusted model indicating that non-genetic factors such as age and sex may be responsible for the association. However, independent studies need to validate our findings with a larger group of NAFLD patients, as well as in different ethnic cohorts.

Lipase Immobilization in Mesoporous Silica Nanoparticles for Biofuel Production

Lipases are ubiquitous enzymes whose physiological role is the hydrolysis of triacylglycerol into fatty acids. They are the most studied and industrially interesting enzymes, thanks to their versatility to promote a plethora of reactions on a wide range of substrates. In fact, depending on the reaction conditions, they can also catalyze synthesis reactions, such as esterification, acidolysis and transesterification. The latter is particularly important for biodiesel production. Biodiesel can be produced from animal fats or vegetable oils and is considered as a biodegradable, non-toxic and renewable energy source. The use of lipases as industrial catalysts is subordinated to their immobilization on insoluble supports, to allow multiple uses and use in continuous processes, but also to stabilize the enzyme, intrinsically prone to denaturation with consequent loss of activity. Among the materials that can be used for lipase immobilization, mesoporous silica nanoparticles represent a good choice due to the combination of thermal and mechanical stability with controlled textural characteristics. Moreover, the presence of abundant surface hydroxyl groups allows for easy chemical surface functionalization. This latter aspect has the main importance since lipases have a high affinity with hydrophobic supports. The objective of this work is to provide an overview of the recent progress of lipase immobilization in mesoporous silica nanoparticles with a focus on biodiesel production.

The influence of oriented external electric field on lipase catalyzed triglyceride hydrolysis

The use of microbial lipases for the hydrolysis of natural oils such as triglyceride esters is a green alternative to conventional processes to produce high value-added chemicals. Biocatalysis using lipases in aqueous solutions is limited often by instabilities, slow kinetics due to mass transfer limitations, and by challenges of economic lipase recycling. Prior work has shown that mass transfer rates and reaction rates can be intensified by increasing interfacial area using electrostatic spray reactors. However, the possible effect of oriented external electrical field on the lipase catalytic activity has hitherto not been considered. In this study, the performance of microbial lipase derived from Candida rugosa in aqueous solutions was analyzed in the presence of a steady DC externally applied electrical voltage. The reaction was conducted in three different batch type reactors, quiescent (fixed interface), stirred tank, and tubular flow system. In each of the three reactor systems it was concluded that the oriented external electrical field has a positive effect on reaction independently of interfacial area. Further studies conducted using reverse polarity, increased electrode distance and for an immobilized lipase system have shown that lipase undergoes conformational changes due to an oriented external electrical field is the main driving mechanism for this enhanced performance.

TRANSESTERIFICATION OF VEGETABLE OILS’ TRIGLYCERIDES BY THE HEAD FRACTION OF ETHYL ALCOHOL

The regularities of sunflower, rapeseed and flaxseed oils’ triglycerides transesterification by the head fraction of ethyl alcohol in the presence of various catalysts have been investigated. The cation exchange resin KU-2-8 in H+-form and cation exchange resin KU-2-8 containing an immobilized Cu2+-ions (untreated and treated by 0,1 M sodium hydroxide solution for 90 min and for 7 days) were used as catalyst. It was shown that the all investigated catalysts except the untreated cation exchange resin KU-2-8 containing an immobilized Cu2+-ions allow to achieve of triglycerides conversion over 90 % under the reaction conditions. It was found that the catalyst KU-2-8 containing an immobilized Cu2+-ions treated by 0.1 M sodium hydroxide solution for 90 min shows the highest activity. Also it was found that this catalyst allows to achieve triglyceride conversion all of studied vegetable oils over 99.5% in 180 min of reaction. Simultaneously it was found that the highest rate of transesterification reaction in its presence is observed. The influence of these catalysts on the esterification reaction of free fatty acids, present in vegetable oils, by ethyl alcohol, present in the head fraction, was studied. It was established that the influence of the investigated catalysts on the esterification reaction is different. It was shown that cation exchange resin KU-2-8 containing an immobilized Cu2+-ions treated by 0.1 M sodium hydroxide solution for 90 min catalyzes the esterification reaction of free fatty acids of all investigated oils. Their conversion is from 21.6 % (in sunflower oil) up to 74.1 % (in rapeseed oil). At the same time, it was found that long-term treatment of cation exchange resin KU-2-8 containing an immobilized Cu2+-ions by sodium hydroxide solution leads to a sharp decrease or loss of catalyst activity in the esterification reaction of free fatty acids by ethyl alcohol. It was shown that the use of this catalyst for the transesterification of rapeseed and sunflower oils’ triglycerides even increases the acid number of the reaction mixture, namely to the partial hydrolysis of triglycerides. The obtained results show that the short-term treatment the cation exchange resin KU-2-8 containing an immobilized Cu2+-ions by 0.1 M alkali solution is one of the ways that allow to increase its catalytic activity in the transesterification process of vegetable oils by head fraction of ethyl alcohol. It is assumed that such treatment affects the catalytic activity of Cu2+-ions due to changes in the electron density on the catalyst cation caused the coordination of part of the OH–-anions. The long-term treatment of the cation exchange resin containing an immobilized Cu2+-ions by alkali solution causes a maximum saturation of the cation coordination sphere. This obviously creates steric problem of a reagents access to the catalyst and reduces the catalytic activity cation exchange resin.

POWER RATE LAW BASED CHEMICAL KINETICS AND THERMODYNAMIC MODELING OF AFRICAN PEAR SEED OIL CONSECUTIVE IRREVERSIBLE BASE METHANOLYSIS FOR BIODIESEL PRODUCTION

Currently the major challenge of biodiesel application as a replacement to petrodiesel is its industrial production sustainability.Consequently, the successful scale-up of laboratory results in transesterification requires so much information obtained through chemical kinetics.This paper presents the kinetics and thermodynamic study of alkali-homogeneous irreversible methanolysis of seed oil derived from African pear. The transesterification process was carried out from 0-100 minutes at temperature range of 55-65°C. The reaction mixture compositions were ascertained using gas chromatography- flame ionization detector (GC-FID) technique. Rate constants of the triglyceride (Tg), diglycerides (Dg) and monoglycerides(Mg) hydrolysis were in the range of 0.0140- 0.07810 wt%/min and increased with increase in temperature. The rate of reaction was found to increase with increase in temperature. Activation energies were found to be 6.14, 20.01 and 28.5kcal/mol at 55, 60 and 65oC respectively. Tg hydrolysis to Dg was observed asthe rate determining step while the reaction agreed with second order principles. A biodiesel yield of 93.02% was obtained with cloud point of 10°C , flash point of 125°C , pour point of 4°C , calorific value of 34.4MJ/kg, and cetane number of 54.90 which satisfy EN14214 and ASTM D 6751 standards. Results presented in this report would serve as idealized conditions for industrial scale up of biodiesel production from African pear seed oil. Keywords:Kinetics; methanolysis; rate constants; activation energy; African pear seed oil; biodiesel