Addition Of Saturated Fatty Acids Research Articles

Let's get fat: emergence of S-acylation as a therapeutic target in Huntington disease.

Protein mislocalization is a key initial step in neurodegeneration, regardless of etiology, and has been linked to changes in the dynamic addition of saturated fatty acids to proteins, a process known as S-acylation. With the advent of new techniques to study S-acylation and the recent discovery of new enzymes that facilitate protein deacylation, novel small molecules are emerging as potential new therapeutic treatments. Huntington disease (HD) is a devastating, fatal neurodegenerative disease characterized by motor, cognitive, and psychiatric deficits caused by a CAG repeat expansion in the HTT gene. The protein that is mutated in HD, huntingtin, is less S-acylated which is associated with mutant HTT aggregation and cytotoxicity. Recent exciting findings indicate that restoring S-acylation in HD models using small molecule inhibitors of the deacylation enzymes is protective. Herein, we set out to describe the known roles of S-acylation in HD and how it can be targeted for therapeutic design.

Optimized Incorporation of Alkynyl Fatty Acid Analogs for the Detection of Fatty Acylated Proteins using Click Chemistry

Fatty acylation, the covalent addition of saturated fatty acids to protein substrates, is important in regulating a myriad of cellular functions in addition to its implications in cancer and neurodegenerative diseases. Recent developments in fatty acylation detection methods have enabled efficient and non-hazardous detection of fatty acylated proteins, particularly through the use of click chemistry with bio-orthogonal labeling. However, click chemistry detection can be limited by the poor solubility and potential toxic effects of adding long chain fatty acids to cell culture. Described here is a labeling approach with optimized delivery using saponified fatty acids in combination with fatty-acid free BSA, as well as delipidated media, which can improve detection of hard to detect fatty acylated proteins. This effect was most pronounced with the alkynyl-stearate analog, 17-ODYA, which has been the most commonly used fatty acid analog in click chemistry detection of acylated proteins. This modification will improve cellular incorporation and increase sensitivity to acylated protein detection. In addition, this approach can be applied in a variety of cell types and combined with other assays such as pulse-chase analysis, stable isotope labeling with amino acids in cell culture, and mass spectrometry for quantitative profiling of fatty acylated proteins.

Optimized Incorporation of Alkynyl Fatty Acid Analogs for the Detection of Fatty Acylated Proteins using Click Chemistry.

Fatty acylation, the covalent addition of saturated fatty acids to protein substrates, is important in regulating a myriad of cellular functions in addition to its implications in cancer and neurodegenerative diseases. Recent developments in fatty acylation detection methods have enabled efficient and non-hazardous detection of fatty acylated proteins, particularly through the use of click chemistry with bio-orthogonal labeling. However, click chemistry detection can be limited by the poor solubility and potential toxic effects of adding long chain fatty acids to cell culture. Described here is a labeling approach with optimized delivery using saponified fatty acids in combination with fatty-acid free BSA, as well as delipidated media, which can improve detection of hard to detect fatty acylated proteins. This effect was most pronounced with the alkynyl-stearate analog, 17-ODYA, which has been the most commonly used fatty acid analog in click chemistry detection of acylated proteins. This modification will improve cellular incorporation and increase sensitivity to acylated protein detection. In addition, this approach can be applied in a variety of cell types and combined with other assays such as pulse-chase analysis, stable isotope labeling with amino acids in cell culture, and mass spectrometry for quantitative profiling of fatty acylated proteins.

Coupling Phase Behavior of Fatty Acid Containing Membranes to Membrane Bio-Mechanics.

Biological membranes constantly modulate their fluidity for proper functioning of the cell. Modulation of membrane properties via regulation of fatty acid composition has gained a renewed interest owing to its relevance in endocytosis, endoplasmic reticulum membrane homeostasis, and adaptation mechanisms in the deep sea. Endowed with significant degrees of freedom, the presence of free fatty acids can alter the curvature of membranes which in turn can alter the response of curvature sensing proteins, thus defining adaptive ways to reconfigure membranes. Most significantly, recent experiments demonstrated that polyunsaturated lipids facilitate membrane bending and fission by endocytic proteins – the first step in the biogenesis of synaptic vesicles. Despite the vital roles of fatty acids, a systematic study relating the interactions between fatty acids and membrane and the consequent effect on the bio-mechanics of membranes under the influence of fatty acids has been sparse. Of specific interest is the vast disparity in the properties of cis and trans fatty acids, that only differ in the orientation of the double bond and yet have entirely unique and opposing chemical properties. Here we demonstrate a combined X-ray diffraction and membrane fluctuation analysis method to couple the structural properties to the biophysical properties of fatty acid-laden membranes to address current gaps in our understanding. By systematically doping pure dioleoyl phosphatidylcholine (DOPC) membranes with cis fatty acid and trans fatty acid we demonstrate that the presence of fatty acids doesn’t always fluidize the membrane. Rather, an intricate balance between the curvature, molecular interactions, as well as the amount of specific fatty acid dictates the fluidity of membranes. Lower concentrations are dominated by the nature of interactions between the phospholipid and the fatty acids. Trans fatty acid increases the rigidity while decreasing the area per lipid similar to the properties depicted by the addition of saturated fatty acids to lipidic membranes. Cis fatty acid however displays the accepted view of having a fluidizing effect at small concentrations. At higher concentrations curvature frustration dominates, leading to increased rigidity irrespective of the type of fatty acid. These results are consistent with theoretical predictions as detailed in the manuscript.

The acyltransferase Gpc1 impacts PC molecular species and phenotypic outcomes

Phospholipids are major components of cellular membranes and their biochemical properties, such as fatty acid composition, help define the biophysical properties of membranes. Phosphatidylcholine (PC) is the most abundant phospholipid in cellular membranes and is synthesized primarily by two well defined metabolic pathways in Saccharomyces cerevisiae. Our laboratory recently characterized a novel glycerophosphocholine (GPC) acyltransferase, encoded by GPC1, that provides a third route for PC biosynthesis and is involved in the newly defined PC deacylation‐reacylation remodeling pathway (PC‐DRP). PC‐DRP begins with GPC, the product of complete PC deacylation by phospholipases of the B type. Gpc1 acylates GPC to form lysophosphatidylcholine (LysoPC). LysoPC can subsequently be converted to PC by Ale1. Previous results have shown that Gpc1 prefers the addition of saturated fatty acids during PC remodeling, which results in more saturated PC species and has the potential to influence the biophysical properties of the membrane. To better understand how GPC1 is regulated and to determine its physiological role in the cell, we assessed the effect of GPC1 dosage and temperature alterations on growth, expression, and PC molecular species. Preliminary data indicates that while increasing the temperature from 30°C to 37°C slows growth in a wild type strain, the gpc1Δ mutant is much less effected. As expected, overexpression of GPC1 further negatively impacts growth at 37°C. GPC1 message abundance, as measured by qRT‐PCR, is decreased at 37°C as compared to 30°C in a wild type strain. Other members of PC‐DRP, including ALE1 and PLB1 also display decreased expression upon elevated temperature. Mass spectrometry‐based analysis of the PC molecular species under these conditions are ongoing. Other stressors expected to impact membrane events, including inositol availability, are being examined. These experiments will provide insight into the physiological role of Gpc1.Support or Funding InformationNIH R15 GM104876 (to JPV)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Huntingtin interacting proteins 14 and 14-like are required for chorioallantoic fusion during early placental development

Huntington disease (HD) is an adult-onset neurodegenerative disease characterized by motor, cognitive, and psychiatric symptoms that is caused by a CAG expansion in the HTT gene. Palmitoylation is the addition of saturated fatty acids to proteins by DHHC palmitoylacyl transferases. HTT is palmitoylated by huntingtin interacting proteins 14 and 14-like (HIP14 and HIP14L or ZDHHC17 and 13 respectively). Mutant HTT is less palmitoylated and this reduction of palmitoylation accelerates its aggregation and increases cellular toxicity. Mouse models deficient in either Hip14 (Hip14−/−) or Hip14l (Hip14l−/−) develop HD-like phenotypes. The biological function of HTT palmitoylation and the role that loss of HTT palmitoylation plays in the pathogenesis of HD are unknown. To address these questions mice deficient for both genes were created. Loss of Hip14 and Hip14l leads to early embryonic lethality at day embryonic day 10–11 due to failed chorioallantoic fusion. The chorion is thickened and disorganized and the allantois does not fuse correctly with the chorion and forms a balloon-like shape compared to Hip14l−/−; Hip14+/+ littermate control embryos. Interestingly, the Hip14−/− ; Hip14−/− embryos share many features with the Htt−/− embryos, including folding of the yolk sac, a bulb shaped allantois, and a thickened and disorganized chorion. This may be due to a decrease in HTT palmitoylation. In Hip14−/−; Hip14l−/− mouse embryonic fibroblasts show a 25% decrease in HTT palmitoylation compared to wild type cells. This is the first description of a double PAT deficient mouse model where loss of a PAT or multiple PATs results in embryonic lethality in mammals. These results reinforce the physiological importance of palmitoylation during embryogenesis.

Formation of Inverse Topology Lyotropic Phases in Dioleoylphosphatidylcholine/Oleic Acid and Dioleoylphosphatidylethanolamine/Oleic Acid Binary Mixtures

The addition of saturated fatty acids (FA) to phosphatidylcholine lipids (PC) that have saturated acyl chains has been shown to promote the formation of lyotropic liquid-crystalline phases with negative mean curvature. PC/FA mixtures may exhibit inverse bicontinuous cubic phases (Im3m, Pn3m) or inverse topology hexagonal phases (HII), depending on the length of the acyl chains/fatty acid. Here we report a detailed study of the phase behavior of binary mixtures of dioleoylphosphatidylcholine (DOPC)/oleic acid (OA) and dioleoylphosphatidylethanolamine (DOPE)/oleic acid at limiting hydration, constructed using small-angle X-ray diffraction (SAXD) data. The phase diagrams of both systems show a succession of phases with increasing negative mean curvature with increasing OA content. At high OA concentrations, we have observed the occurrence of an inverse micellar Fd3m phase in both systems. Hitherto, this phase had not been reported for phosphatidylethanolamine/fatty acid mixtures, and as such it highlights an additional route through which fatty acids may increase the propensity of bilayer lipid membranes to curve. We also propose a method that uses the temperature dependence of the lattice parameters of the HII phases to estimate the spontaneous radii of curvature (R0) of the binary mixtures and of the component lipids. Using this method, we calculated the R0 values of the complexes comprising one phospholipid molecule and two fatty acid molecules, which have been postulated to drive the formation of inverse phases in PL/FA mixtures. These are -1.8 nm (±0.4 nm) for DOPC(OA)2 and -1.1 nm (±0.1 nm) for DOPE(OA)2. R0 values estimated in this way allow the quantification of the contribution that different lipid species make to membrane curvature elastic properties and hence of their effect on the function of membrane-bound proteins.

Lipid droplet formation protects against gluco/lipotoxicity in Candida parapsilosis: An essential role of fatty acid desaturase Ole1

Elevated levels of glucose and lipids can result in cellular dysfunction in eukaryotic cells ranging from Saccharomyces cerevisiae yeasts to human cells. Moreover, glucotoxicity and lipotoxicity can cause cell death, although the mechanism(s) for lethality is unclear. In the present study, we utilized Candida parapsilosis fatty acid desaturase (OLE1) and fatty acid synthase (FAS2) gene deletion mutants and wild-type (WT) yeast cells to unravel the relationship to glucose and lipid induced cell death in eukaryotic cells. Incubation of WT yeast cells with glucose led to the rapid accumulation of lipid droplets, whereas lipid droplet formation was severely impaired in yeast cells with deletion of OLE1 (ole1Δ/Δ) or FAS2 (fas2Δ/Δ). Interestingly, ole1Δ/Δ yeast cells died within hours in a 1% glucose medium without fatty acid supplementation, whereas the WT or fas2Δ/Δ yeast cells did not. In glucose medium, ole1Δ/Δ yeast cells accumulated saturated fatty acids, while fas2Δ/Δ did not. Addition of saturated fatty acids (e.g., palmitic acid) enhanced ole1Δ/Δ yeast cell death, whereas the addition of unsaturated fatty acids (e.g., oleic or palmitoleic acid) rescued cell death. Furthermore, palmitic acid and glucose medium induced apopotic cell death in ole1Δ/Δ yeast cells, which was dependent on mitochondrial function. Thus, our results show that glucotoxicity is directly linked to lipotoxicity, which we demonstrate is mediated by mitochondrial function.

Saturated fatty acids suppress adrenocorticotropic hormone (ACTH) release from rat anterior pituitary cells in vitro

We studied whether fatty acids modify adrenocorticotropic hormone (ACTH) release induced by stimulation with corticotropin-releasing hormone (CRH) from rat anterior pituitary cells. Stimulation with CRH (0.01–100 nmol/l) significantly and concentration-dependently increased ACTH release, which was synergistically enhanced by the simultaneous stimulation with 1 nmol/l arginine-vasopressin. Addition of saturated fatty acids (butyrate, caprylate, laurate, palmitate and stearate) in a medium at 1 mmol/l, despite effects on the basal release, significantly reduced the ACTH release induced by CRH (1 nmol/l) stimulation. Caprylate suppressed ACTH release in a concentration-dependent manner. However, unsaturated C18 and C20 fatty acids (oleate, linolate, linolenate and arachidonate) at 1 mmol/l significantly increased the basal release, but none of them suppressed CRH (1 nmol/l)-induced ACTH release. In the presence of caprylate (1 mmol/l), CRH (1 nmol/l)-stimulated increase in cellular calcium ion concentration was diminished. From these results we conclude that saturated fatty acids have a suppressing effect on CRH-induced ACTH increase in primary cultured rat anterior pituitary cells.

Ruminal and total tract digestibilities in steers fed diets containing liquefied or prilled saturated fatty acids.

We studied the effects of the addition of liquefied vs prilled mostly saturated fatty acids (FA) to the concentrate portion of total mixed diets on ruminal and total tract digestibilities. Four Holstein steers (270 +/- 23 kg) fitted with ruminal and duodenal cannulas were used in a 4 x 4 Latin square design with 21-d periods. Diets contained (DM basis) 30.0% corn silage, 22.2% chopped alfalfa hay, 25.0% ground shelled corn, 12.5% soybean meal, and 5% of one of the following fat sources: 1) prilled FA (PFA), 2) choice white grease (triglycerides) added in liquid form (LTG), 3) 2.5% PFA + 2.5% LTG, or 4) 2.5% liquefied PFA (LFA) + 2.5% LTG. Ad libitum OM intake was not different (P > .10) among diets (mean 7.8 kg/d). Ruminal digestibilities of OM (35.7, 39.9, 42.2, and 37.3% for Diets 1 to 4, respectively) were greatest (P < .10) for the combination of PFA + LTG and lowest for PFA alone. Ruminal digestibilities of NDF, ADF, and starch did not differ (P > .10) among diets. Total tract digestibilities of OM, NDF, and ADF were greater (P < .10) for the diet containing LTG alone than for the diet containing LFA + LTG because of trends for greater postruminal digestibilities. The LFA + LTG diet resulted in a greater proportion of acetate and lower proportion of propionate in ruminal fluid than PFA alone (P < .10). The acetate:propionate ratio (3.53, 2.96, 3.10, and 2.89 for Diets 1 to 4, respectively) was lower (P < .05) for LFA + LTG or LTG alone than for PFA alone. Postruminal and total tract digestibilities of total FA (66.0, 76.0, 71.2, and 68.9% for Diets 1 to 4, respectively) were lower (P < .05) for PFA than for other diets. Addition of saturated FA in liquid form resulted in digestibilities and ruminal effects similar to the same saturated FA added in prilled form.

天然ゴムの結晶化挙動の解析 （第２報） 結合脂肪酸の影響

The effect of fatty acid linked to cis-1, 4 polyisoprene on the crystallization of polymer chain was investigated by differential scanning calorimetry (DSC) and dilatometry at -25°C. Synthetic cis-1, 4 polyisoprene (IR) was esterified with stearic acid selectively at 3, 4 isoprene units after introduction of OH group by hydroboration (IR-C 18). The rate of crystallization of IR-C 18 was increased by the addition of saturated fatty acids. The addition of methyl linoleate promoted the crystallization of IR-C 18, whereas it suppressed the crystallization of IR. This was explained to be due to a synergistic effect of mixed and linked fatty acids on the crystallization of IR. Stearic acid was introduced into natural rubber (NR) by esterification after purification of NR by deproteinization followed by acetone extraction and transesterification with NaOCH3. The content of stearic acid in the esterified NR (DPNR-C 18) was about 1/2 compared to the ester content in untreated NR. The rate of crystallization of DPNR-C 18 was similar to that of DPNR-TE and was not increased by the addition of methyl linoleate. Precooling of the rubber at 0, 10 and 30°C before crystallization at -25°C resulted in a significant difference in the crystallization rate between untreated NR, DPNR-TE and DPNR-C 18, where the crystallization of only untreated NR was promoted by precooling. This demonstrates the presence of a critical concentration of linked fatty acid on the crystallization of NR, at which the synergistic effect was promoted with mixed fatty acids.

Modification of Milkfat Physical Properties by ImmobilizedPseudomonas fluorescensLipase

A bifunctional fusion protein was constructed by fusion of the streptavidin gene from Streptomyces avidinii to the lipase gene from Pseudomonas fluorescens, and the resulting streptavidin−lipase was expressed in Escherichia coli. Immobilized streptavidin−lipase was prepared by direct bioselective adsorption from crude cell lysates on biotinylated controlled-pore glass and used to catalyze interesterification of anhydrous butteroil. Changes in the triacylglycerol composition indicated that those with equivalent carbon numbers (ECN) ranging from 36 to 42 decreased, while those with ECN values from 48 to 50 increased following interesterification for 120 h in hexane at 42 °C. Both the melting temperatures and the solid fat content at various temperatures were lower as compared to those of the unmodified butteroil. Addition of unsaturated fatty acids, linoleic and linolenic, yielded modified butteroils with lower melting points and solid fat content, whereas addition of saturated fatty acids, palmitic and ste...

The effects of dietary fatty acids on the proliferation of normal human urothelial cells in vitro.

Little is known of the mechanisms by which dietary fatty acids (FAs) may affect normal epithelial cell physiology and thereby directly or indirectly influence tumour incidence and progression. In this study, we have used normal human urothelial cell cultures to investigate whether FAs may modify proliferation of normal human epithelial cells in vitro. FAs were presented as albumin complexes in serum-free medium and the effects on proliferation over a concentration range of 1-100 microM were assayed by thymidine incorporation. Saturated FAs had no effect. At lower concentrations (1-10 microM), mono-unsaturated FAs (MUFAs) and n-3 polyunsaturated FAs (PUFAs) were slightly stimulatory. Concentrations of unsaturated FAs above 10 microM were growth inhibitory in a dose-dependent manner. Oleic acid showed least cytostatic effect, whereas gamma-linolenic acid induced irreversible growth arrest. Although marked morphological effects were observed in irreversibly growth-inhibited cells, the cells remained attached to the substratum and showed no evidence of nuclear pyknosis or apoptosis. The inhibitory effects of single PUFAs could be reduced, but not abolished, by the addition of saturated FAs or MUFAs. Mixtures of different PUFAs were inhibitory in an additive manner. These data suggest that PUFAs have a cytostatic effect on rapidly proliferating epithelial cells which appears unrelated to malignant transformation.

Fatty acylation of heparan sulfate proteoglycan from human colon carcinoma cells.

A number of transmembrane proteins have been recently reported to be modified by the covalent addition of saturated fatty acids which may contribute to membrane targeting and specific protein-lipid interactions. Such modifications have not been reported in cell-associated heparan sulfate proteoglycans, although these macromolecules are known to be hydrophobic. Here, we report that a cell surface heparan sulfate proteoglycan is acylated with both myristate and palmitate, two long-chain saturated fatty acids. When colon carcinoma cells were labeled with [3H]myristic acid, a significant proportion of the label was shown to be specifically incorporated into the protein core of the proteoglycan. Characterization of fatty acyl moiety in the purified proteoglycan by reverse-phase high pressure liquid chromatography revealed that approximately 60% of the covalently bound fatty acids was myristate. We further show that this relatively rare 14-carbon fatty acid was bound to the protein core via a hydroxylamine- and alkali-resistant amide bond. The remaining 40% was the more common 16-carbon palmitate, which was bound via a hydroxylamine- and alkali-sensitive thioester bond. Palmitate appeared to be added post-translationally and derived in part from intracellular elongation of myristate, a process that occurred within the first two hours and was insensitive to inhibition of protein synthesis. Acylation of heparan sulfate proteoglycan represents a novel modification of this gene product and could play a role in a number of biological functions including specific interactions with membrane receptors and ligand stabilization.

Lipids and Rheological Properties of Starch Part I: The Effect of Fatty Acids, Monoglycerides and Monoglyceride Ethers on Pasting Temperature and Viscosity of Wheat Starch

AbstractBy addition of saturated fatty acids (C14‐C18) in concentrations of 0.34 and 0.57% gelatinization temperature (GT) of wheat starch is reduced from 82.5°C to 81°C. Oleic acid (C18:1) increases GT. Palmitic and stearic acid (0.57%) cause a little increase of viscosity only, but during heating a slope of viscosity is observed. In contrast to saturated fatty acids monoglyderides increase GT with exception of glycerinmonooleate, which reduces GT. Glycerinmonomyristate and glycerinmonolinoleate increase the initial viscosity. Monoglyceride ethers, first time used for those studies, influence viscosity differently. Rac‐1‐O‐octadecylglycerol reduces GT from 82.5 to 81°C and the final viscosity is lowered remarkable but rac‐1‐O‐[(Z)‐9‐octadecenyl]glycerol causes an increase of GT from 82.5 to 85.5°C. Initial and end viscosity are raised also.

In vitro oxidation of alpha-tocopherol (vitamin E) in human platelets upon incubation with unsaturated fatty acids, diamide and superoxide.

Incubation of human blood platelets in vitro in Tyrode solution with unsaturated fatty acids, diamide or superoxide (generated in situ) resulted in the oxidation of tocopherol in the platelets. Arachidonate concentrations of (3-5).10(-4) M caused a 50% decrease in platelet alpha-tocopherol. The addition of saturated fatty acids or platelet-active substances such as ADP, dibutyryl cyclic AMP, and some prostaglandins, or peroxidizing agents such as hydrogen peroxide and tert-butylhydroperoxide to the incubation medium did not cause any change in platelet tocopherol content. During incubations of platelets with arachidonate, malonaldehyde as well as alpha-tocopherolquinone were produced. The latter was also produced during incubations with diamide or superoxide. The oxidation of tocopherol induced by unsaturated fatty acids may be one factor responsible for the well-known increase in dietary vitamin E requirements induced by polyunsaturated fatty acids. The oxidative consumption of tocopherol in the membranes could be expected to take place during localized release of oxidants such as superoxide and polyunsaturated fatty acids during normal biological function (e.g., phagocytosis) or pathological processes (e.g., ischemia). Tocopherol utilization is kept low probably by the regeneration of the compound by vitamin C and/or the preferential utilization of the other biological antioxidants.

Influence of diet on the incorporation of radioactive fatty acids into the glycerolipids during development of the insectCeratitis capitata

Abstract 1. Larvae ofCeratitis capitata were reared on three different lipid-containing diets. The levels of incorporation of palmitic and linoleic acids into triacylglycerols, phosphatidylethanolamine and phosphatidylcholine, from larvae and pharate adults, were determined. 2. The modified diets cause an increase in the incorporation of linoleic and palmitic acids into the larvae triacylglycerols and pharate adult phospholipids. 3. The addition of saturated fatty acids to the diet cause a higher incorporation of palmitic acid, than that of linoleic acid, into the larvae and pharate adult triacylglycerols. 4. In pharate adults coming from larvae reared on modified diets, there is a higher incorporation of linoleic acid into phospholipids compared with those of larvae. 5. These results indicate that dietary fatty acids have an influence on larvae triacylglycerol and pharate adults phospholipid biosynthesis.

Improvement of the productivity of elasnin, a specific elastase inhibitor, by Streptomyces noboritoensis KM-2753.

Streptomyces noboritoensis KM-2753, an elasnin-producing strain, co-produces an antimycin complex and its elasnin productivity is low (0.006 mg/ml). To obtain mutants possessing higher degrees of elasnin productivity and deficient in antimycin production, the strain was treated with N-methyl-N'-nitro-N-nitrosoguanidine (NTG), ultraviolet light, acriflavine (AF) and high temperature. Mutant N-134 obtained by treatment with NTG was 108 times higher than the original strain in elasnin productivity and produced no antimycin. Strain AF-17 obtained by AF treatment and strain H-80 obtained by incubation at high temperature showed 715 and 428 times higher productivities than that of the original strain, respectively. The productivity of elasnin was further increased 1.6 approximately 2.6 times by addition of saturated fatty acids, especially lauric acid, to the culture. Consequently, strain AF-17 produced 11.1 mg/ml of elasnin in the presence of 10 microgram/ml of lauric acid.