Influence of vegetable oils fatty acid composition on reaction temperature and glycerides conversion to biodiesel during transesterification

Presence of unreacted glycerides in biodiesel may reduce drastically its quality. This is why conversion of raw material in biodiesel through transesterification needs to readjust reaction parameter values to complete. In the present work, monitoring of glycerides transformation in biodiesel during the transesterification of vegetable oils was carried out. To check the influence of the chemical composition on glycerides conversion, selected vegetable oils covered a wide range of fatty acid composition. Reactions were carried out under alkali-transesterification in the presence of methanol. In addition, a multiple regression model was proposed. Results showed that kinetics depends on chemical and physical properties of the oils. It was found that the optimal reaction temperature depends on both length and unsaturation degree of vegetable oils fatty acid chains. Vegetable oils with higher degree of unsaturation exhibit faster monoglycerides conversion to biodiesel. It can be concluded that fatty acid composition influences reaction parameters and glycerides conversion, hence biodiesel yield and economic viability.     

A study of the fatty acid metabolism of the yeast Pityrosporum ovale

The yeast Pityrosporum ovale, a skin saprophyte, will only grow if fatty acids of chain length greater than C(10) are added to the culture medium. 9-Hydroxypalmitic acid is the major product of metabolism of even-carbon-number fatty acids; 9-hydroxystearic acid is also found. The optimum pH for this conversion is pH4.5. The hydroxy fatty acids produced are found bound in a polar form in the aqueous phase of the culture medium. Growth of the organisms is facilitated by presentation of the substrate as a two-phase liquid system.     

Effect of Substrate on the Fatty Acid Composition of Hydrocarbon- and Ketone-utilizing Microorganisms

The fatty acid pattern in hydrocarbon- and ketone-utilizing bacteria after growth on various substrates was examined. The fatty acid composition of one hydrocarbon-utilizing organism (Mycobacterium sp. strain OFS) was investigated in detail after growth on n-alkanes, 1-alkenes, ketones, and n-alcohols. n-Alkanes shorter than C₁₃ or longer than C₁₇ were not incorporated into cellular fatty acids without some degradation. Strain OFS incorporated C₁₄ to C₁₇ 1-alkenes into cellular fatty acids as the ω-monoenoic fatty acid. Methyl ketones were incorporated into strain OFS after removal of one- or two-carbon fragments from the carbonyl end of the molecule. An organism isolated by enrichment on methyl ketones was incapable of n-alkane utilization but could grow on, although not incorporate, ketones or long chain n-alcohols into cellular fatty acids.     

The influence of carbohydrate dissimilation on the fatty acid metabolism of Monascus purpureus

The formation of methyl ketones from fatty acids is known as a detoxifying mechanism and takes place during the lag period of growth. Fermentations of fatty acid and saccharides mixture led to two-phase growth kinetics. It was found that first the fatty acid is oxidized before saccharide dissimilation and growth occurs. This principle can also be observed in fermentation processes with precursor mixtures. Precursor-induced breakdown happens due to the toxicity of fatty acids and methyl ketones. Thereby the fatty acid is metabolized prior to the methyl ketone. After this, saccharide dissimilation and growth take place. Correspondence to: N. Peters     

Fatty acid composition of Bifidobacterium and Lactobacillus strains

Normal C(14), C(16), and C(18) saturated acids and C(16) and C(18) monoenoic acids are the main fatty acids of nine strains of Bifidobacterium. Their lactobacillic acid content was less than 5%. Lactobacillus strains contained the same fatty acids as main compounds except for octadecanoic acid, which was present only in very low amounts. Eight of nine Lactobacillus strains contained in the stationary phase more than 15% lactobacillic acid. No correlation was observed between the fatty acid composition and other physiological characteristics used in the literature for classification of strains of one genus. Aging of the culture, which involved a decrease of the pH, caused a lengthening of the chain length of the fatty acids of B. bifidum var. pennsylvanicus but only a conversion of octadecenoic to lactobacillic acid in the lactobacilli. Lowering of the temperature of cultivation decreased the chain length of the fatty acids of B. bifidum var. pennsylvanicus. L. lactis did not show any influence of the temperature on the chain length of the fatty acids. The percentage of unsaturated acids was temperature independent in both organisms.     

Glyceride synthesis by four kinds of microbial lipase

Apart from their usual mechanism of action, lipases from Aspergillus niger and Rhizopus delemar also catalyzed the synthesis of glycerides from oleic acid and glycerol. Lipases from Geotrichum candidum and Penicillium cyclopium were inactivated by oleic acid, but were stable in the presence of casein, albumin or buffer of appropriate pH. Lipases from Aspergillus niger and Rhizopus delemar synthesized glycerides from, not only fatty acid, but dibasic acids and aromatic acids, making ester bonds only at position 1 and 3 of glycerol. In contrast, lipases from Geotricum candidum and Penicillium cyclopium synthesized glycerides only from long chain fatty acids, and made ester bonds at all three available positions of the glycerol molecule.     

Biotransformation of free fatty acids in mixtures to methyl ketones by Monascus purpureus

Summary Monascus purpureus converts short-chain fatty acids to methyl ketones. The regulation of the metabolic pathway is similar to that found in Penicillium roquefortii. There are differences in the actual amount of precursors metabolized. The fermentation of fatty acid mixtures led to methyl ketone mixtures. The metabolism of each fatty acid was dependent on the precursor composition. Offprint requests to: C. Kranz     

The fatty acid composition of sphingolipids from bovine CNS axons and myelin

Abstract— Cerebrosides, sulphatides and sphingomyelin were isolated from bovine CNS myelin and from myelin-free axons derived from myelinated axons. The fatty acid composition of each sphingolipid was determined by gas-liquid chromatography of the fatty acid methyl esters. In each case the fatty acids of the axonal sphingolipids were of shorter average chain length than those from the corresponding myelin lipids. These differences, however, were small and the fatty acids of the axonal cerebrosides and sulphatides were similar in average chain length to those reported previously for bovine myelin. The principal unsubstituted acid of both cerebroside and sulphatide from axons was 24: 1, with the total long chain acids (> C₁₈) amounting to 80 and 85 per cent, respectively. The corresponding figures for myelin galactolipids were 94 and 95 per cent long chain acids. The principal α-hydroxy acid of both axonal galactolipids was 24 h:0, with cerebroside having 80 per cent and sulphatide 92 per cent long chain acids, compared to the figures of 87 and 97 per cent for the corresponding myelin lipids. In axonal sphingomyelin the major acid was 18:0 (compared to 24:1 in myelin) and the long chain acids were 61 per cent of the total vs 76 per cent of the total for myelin sphingomyelin. The non-identity of axonal and myelin sphingolipid fatty acids substantiates the belief that they are intrinsic axonal constituents. These findings do not rule out the possibility of a close metabolic relationship between the sphingolipids of the axon and its myelin sheath.     

Fatty acid metabolism of isolated mammalian cells

It is now clear that a wide variety of differentiated cells in culture exhibit essentially the full spectrum of mammalian fatty acid metabolism. These cells readily incorporate free fatty acids into membrane phosphoglycerides, modify exogenous fatty acids by desaturation and elongation, and store excess fatty acyl groups, primarily as triacylglycerols. Similarly, many different types of cells synthesize cyclooxygenase and lipoxygenase derivatives of long chain polyunsaturated fatty acids. Furthermore, although the fatty acid composition of cellular phospholipids can be modified by medium supplementation, cells in culture exhibit definite fatty acyl specificities for the various steps of fatty acid activation, transesterification and release. As the extensive repertoire of fatty acid metabolism in mammalian cells has been elucidated, and as the ability to grow differentiated cells in culture has increased, new questions have arisen. There is still much to be learned about the enzymes involved in synthesizing and maintaining the unique fatty acid composition of the different cellular phospholipids and the processes which regulate the desaturation, elongation and retroconversion of polyunsaturated fatty acids. Other areas of great current interest are the mechanisms by which certain long chain polyunsaturated fatty acids are made available for conversion to oxygenated, biologically-active derivatives, the metabolic interactions between different polyunsaturated fatty acids, particularly n-3 and n-6 fatty acids, the cellular roles of the C22 polyunsaturated fatty acids, and the functions of particular molecular species of phospholipids in membrane-mediated events. Further research in these areas will contribute to unravelling the role of fatty acids and fatty acid derivatives in the physiological processes of mammalian cells.     

Identification and characterization of a fatty acid binding protein in bovine mammary gland

A fatty acid binding protein (FABP) was isolated from bovine mammary cytosol by gel filtration and ion exchange chromatography. Polyacrylamide gel electrophoresis in sodium dodecyl sulfate indicated a mol. wt. of 12,000. Isoelectric focusing showed two bands at pH 5.6 and 5.8. FABP bound long chain fatty acids and their CoA thioesters, but not medium or short chain fatty acids. Affinity constant (Ka) for 18:1 was about 2 micromolar. Endogenously bound fatty acids included 16:0, 18:0 and 18:1, in both covalent and noncovalent association with FABP. Activities of microsomal phosphatidic acid phosphatase, fatty acid:CoA ligase or diacylglycerol acyltransferase were not affected by purified FABP in vitro.     

Studies to determine the role rates of chain elongation and desaturation play in regulating the unsaturated fatty acid composition of rat liver lipids.

Rat liver microsomes were used to measure the rates of chain elongation and desaturation of acids in the linoleate, oleate and palmitoleate biosynthetic pathways. These studies were designed to determine whether there is a relationship between rates of conversion and the types of unsaturated fatty acids found in rat liver lipids. In some cases rates of conversion correlate well with the types of unsaturated fatty acid found inrat liver lipids. In other cases, rates of conversion must be correlated with other controls such as competitive interactions, retroconversion, and specificities for incorporating given acids into lipids in order to explain the unsaturated fatty acid composition of rat liver lipids. The roles and interrelationships of these various metabolic processes are discussed relative to the control of polyunsaturated fatty acid biosynthesis.     

Phase behavior of stratum corneum lipids in mixed Langmuir-Blodgett monolayers.

The lipids found in the bilayers of the stratum corneum fulfill the vital barrier role of mammalian bodies. The main classes of lipids found in stratum corneum are ceramides, cholesterol, and free fatty acids. For an investigation of their phase behavior, mixed Langmuir-Blodgett monolayers of these lipids were prepared. Atomic force microscopy was used to investigate the structure of the monolayers as a function of the monolayer composition. Three different types of ceramide were used: ceramide extracted from pigskin, a commercially available ceramide with several fatty acid chain lengths, and two synthetic ceramides that have only one fatty acid chain length. In pigskin ceramide-cholesterol mixed monolayers phase separation was observed. This phase separation was also found for the commercially available type III Sigma ceramide-cholesterol mixed monolayers with molar ratios ranging from 1:0.1 to 1:1. These monolayers separated into two phases, one composed of the long fatty acid chain fraction of Sigma ceramide III and the other of the short fatty acid chain fraction of Sigma ceramide III mixed with cholesterol. Mixtures with a higher cholesterol content consisted of only one phase. These observations were confirmed by the results obtained with synthetic ceramides, which have only one fatty acid chain length. The synthetic ceramide with a palmitic acid (16:0) chain mixed with cholesterol, and the synthetic ceramide with a lignoceric acid (24:0) chain did not. Free fatty acids showed a preference to mix with one of these phases, depending on their fatty acid chain lengths. The results of this investigation suggest that the model system used in this study is in good agreement with those of other studies concerning the phase behavior of the stratum corneum lipids. By varying the composition of the monolayers one can study the role of each lipid class in detail.     

Conversion of Digalactosyl Diglyceride (Extra Long Carbon Chain Conjugates) into Monogalactosyl Diglyceride of Pine Needle Chloroplasts upon Dehardening

Changes in lipid and fatty acid composition of pine needle chloroplasts were determined upon transfer of the trees from outside (-5°C) to 32°C. Within 7 1/2 hours after transfer, conversion of a portion of digalactosyldiglyceride into monogalactosyl diglyceride was observed. This portion consisted of a conjugate with extra long chain fatty acids (cerotic acid, 26: 0, and C₂₆-cyclo-propane fatty acid). Only minor further changes in lipid and fatty acid composition were observed. Dehardening of the needles occurred within the same period. The data support the hypothesis that the degalactosidation reaction specifically depends on the fatty acid composition. The extra long chain fatty acids may link different sub-units of the chloroplasts and may contribute to an additional layer of H-bonded water on the membrane surface because of the increased exposure of the galactose groups.     

Branched long chain bases in cerebrosides of the guinea pig Harderian gland

Cerebrosides obtained from the guinea pig Harderian gland were analyzed. The purified cerebrosides gave a single spot on thin-layer chromatography, the Rf value being similar to that of phrenosine obtained from whale brain. The cerebrosides consisted of 74.7% of glucosylceramide and 25.3% of galactosylceramide. The fatty acid composition of these cerebrosides was 0.7% of non-hydroxy fatty acids and 99.3% of alpha-hydroxy fatty acids. Among these alpha-hydroxy fatty acids, a small amount of methyl branched acids was detected. The substituted position of methyl branching of alpha-hydroxy fatty acids was the 16th carbon atom from the carboxyl end irrespective of the carbon chain length. The long chain bases were composed of sphinganine (78%) and sphingenine (22%). 4-D-Hydroxysphinganine was not found. The most remarkable feature of the long chain bases of cerebrosides in the Harderian gland was the presence of a large amount of methyl branched sphinganine. The cerebrosides obtained from the cerebrum and cerebellum of the same animal were also analyzed. The sugar, fatty acid, and long chain base compositions of these cerebrosides were similar to those of whale brain cerebrosides. Methyl branched sphinganine was not found in guinea pig brain.     

Lipase-catalyzed synthesis of partial glyceride

Summary Mucor miehei (IM 20) and Candida antarctica (SP 382) lipases were used for esterification of free fatty acids in the absence of organic solvent or transesterification of fatty acid methyl esters in hexane with isopropylidene glycerols. Acid catalyzed cleavage of the isopropylidene groups resulted in the formation of monoacyl glycerol (MAG) and diacyl glycerol (DAG). Both oleic (18:1 n-9) and eicosapentaenoic acid, EPA (20:5 n-3) were successfully incorporated into glycerides. Total acyl donor conversion ranged from 46.9 – 96.9% with MAG content of up to 88.5%.     

The effect of temperature, water activity and sorbic acid on ketone rancidity produced by Penicillium crustosum Thom in coconut and palm kernel oils

In a model system medium-chain fatty acids (MCFA) C₆–C₁₂ in coconut and palm kernel oil are converted to methyl ketones, one carbon atom less than the parent fatty acids, by two strains of Penicillium crustosum Thom. Conversion rates of up to 32% were seen for decanoic acid at 25°C. The optimum temperature for ketone production was 25°C in liquid suspension culture. Coconut oil contains 3.23 mmol/g MCFAs compared with 2.34 mmol/g for palm kernel oil. Coconut oil is more prone to fungal spoilage (growth and ketone production) than palm kernel oil. The main end product of fermentation was 2-undecanone reflecting the high concentration of dodecanoic acid in the substrates. Ketonic rancidity is fungal engendered. The reaction can be controlled by reducing the temperature (4°C), reducing the water activity (0.91) or by addition of sorbic acid (20 mmol/l).     

Antifungal properties of 2-alkynoic acids and their methyl esters.

Thirteen 2-alkynoic acids and their methyl esters (C3--C12, C14, C16, and C18) were tested against Aspergillus niger, Trichoderma viride, and Myrothecium verrucaria in Sabouraud dextrose agar at pH 4.0 and 5.6. Toxicity to Candida albicans, Trichophyton mentagrophytes, and Mucro mucedo was determined in the same medium at pH 5.6 and 7.0 in the absence and presence of 10% beef serum. The fungitoxicity of the acids was influenced by chain length, pH of the medium, and absence or presence of adsorbents. The toxicity of the esters was influenced primarily by chain length and to a lesser extent by the pH of the medium and the presence of beef serum. The order of activity of the 2-alkynoic acids is C10=C11=C12 greater than C14=C16 greater than C9 greater than C8 greater than C7. When compared with other fatty acid analogs, the order of fungitoxicity on a weight basis is 2-alkynoic acids greater than 2-alkenoic acids greater than alkanoic acids greater than 2-bromoalkanoic acids greater than 2-fluoroalkanoic acids. There is an inverse relationship between chain length and pKa of the acids, suggesting that partition behavior is a fundamental determinant of fungitoxicity along with the effect of adsorbents.     

Production of medium chain glycerides from coconut and palm kernel fatty acid distillates by lipase-catalyzed reactions

Medium chain glycerides (MCGs) containing C_8:0 and C_10:0 fatty acids is very much important for medicinal and nutritional applications. Coconut and palm kernel fatty acid distillates (FADs) can be utilized to produce MCGs by a combination of lipase-catalyzed hydrolysis and esterification reactions. The neutral glycerides present in coconut and palm kernel FADs are hydrolyzed by Candida rugosa lipase. The hydrolysates were then subjected to steam distillation under vacuum (at 120–140 °C) to get fractions rich in medium chain fatty acids (MCFAs). The fractions, from coconut and palm kernel FADs (75.2 and 76.2% MCFAs, respectively), were esterified with Rhizomucor miehei (Lipozyme RM IM) lipase to produce MCGs. Products from coconut FAD contained 64.7–67.5% diacylglycerol (DG), followed by 18.8–22.9% monoacylglycerol (MG) and 9.8–9.3% triacylglycerol (TG). Similarly, products from palm kernel FAD contained 63.5–66.7% DG, 19.1–23.6% MG and 9.5–10.1% TG.