On the purification of a fatty acid synthetase complex from an insect

1976 ◽  
Vol 54 (8) ◽  
pp. 1397-1399 ◽  
Author(s):  
S. N. Thompson ◽  
J. S. Barlow
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Column Chromatography ◽  
De Novo ◽  
Gel Filtration ◽  
Fatty Acid Synthetase ◽  
De Novo Synthesis ◽  
Anion Exchange Column ◽  
Malonyl Coa ◽  
Salt Fractionation

The fatty acid synthetase complex of the blowfly, Lucilia sericata, catalyzing the de novo synthesis of fatty acids from acetyl- and malonyl-CoA (EC 2.3.1.9 and EC 2.3.1.39 respectively) and requiring NADPH, was isolated from whole-insect homogenates by ultracentrifugation. Purification was carried out by salt fractionation, anion exchange column chromatography on ECTEOLA (epichlorohydrin triethanolamine) cellulose, and gel filtration column chromatography on Sephadex®.

Effect of Thiolactomycin on de novo Fatty Acid Biosynthesis in Plants

1990 ◽  
Vol 45 (5) ◽  
pp. 518-520 ◽  
Author(s):  
Manfred Focke ◽  
Andrea Feld ◽  
Hartmut K. Lichtenthaler
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Fatty Acid Biosynthesis ◽  
De Novo ◽  
Fatty Acid Synthetase ◽  
Acetate Incorporation ◽  
Acid Biosynthesis ◽  
Acetyl Coa ◽  
Malonyl Coa ◽  
Total Fatty Acids

Thiolactomycin was shown to be a potent inhibitor of de novo fatty acid biosynthesis in intact isolated chloroplasts (measured as [14C]acetate incorporation into total fatty acids). In our attempt to further localize the inhibition site we confirmed the inhibition with a fatty acid synthetase preparation, measuring the incorporation of [14C]malonyl-CoA into total fatty acids. From the two proposed enzymic targets of the fatty acid synthetase by thiolactomycin we could exclude the acetyl-CoA: ACP transacetylase. It appears that the inhibition by thiolactomycin occurs on the level of the condensing enzymes, i.e. the 3-oxoacyl-ACP synthases. We also demonstrated that the two starting enzymes of de novo fatty acid biosynthesis, the acetyl-CoA synthetase and the acetyl-CoA carboxylase, are not affected by thiolactomycin.

scholarly journals The fractionation of the fatty acid synthetase activities of avocado mesocarp plastids

1975 ◽  
Vol 146 (2) ◽  
pp. 439-445 ◽  
Author(s):  
P J Weaire ◽  
R G O Kekwick
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
De Novo ◽  
Acyl Carrier Protein ◽  
Fatty Acid Synthetase ◽  
Carrier Protein ◽  
Acetyl Coa ◽  
Principal Product ◽  
Absolute Requirement ◽  
Malonyl Coa

1. The range of fatty acids formed by preparations of ultrasonically ruptured avocado mesocarp plastids was dependent on the substrate. Whereas [1-14C]palmitate and [14C]oleate were the major products obtained from [-14C]acetate and [1-14C]acetyl-CoA, the principal product from [2-14C]malonyl-CoA was [14-C]stearate. 2. Ultracentrifugation of the ruptured plastids at 105000g gave a supernatant that formed mainly stearate from [2-14C]malonyl-CoA and to a lesser extent from [1-14C]acetate. The incorporation of [1-14C]acetate into stearate by this fraction was inhibited by avidin. 3. The 105000g precipitate of the disrupted plastids incorporated [1-14C]acetate into a mixture of fatty acids that contained largely [14C]plamitate and [14C]oleate. The formation of [14C]palmitate and [14C]oleate by disrupted plastids was unaffected by avidin. 4. The soluble fatty acid synthetase was precipitated from the 105000g supernatant in the 35-65%-saturated-(NH4)2SO4 fraction and showed an absolute requirement for acyl-carrier protein. 5. Both fractions synthesized fatty acids de novo.

In vivo study of the biosynthesis of long-chain fatty acids using deuterated water

1995 ◽  
Vol 269 (2) ◽  
pp. E247-E252 ◽  
Author(s):  
H. O. Ajie ◽  
M. J. Connor ◽  
W. N. Lee ◽  
S. Bassilian ◽  
E. A. Bergner ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
De Novo ◽  
Chain Elongation ◽  
Deuterated Water ◽  
De Novo Synthesis ◽  
Isotopomer Distribution ◽  
Long Chain ◽  
Mass Isotopomer

To determine the contributions of preexisting fatty acid, de novo synthesis, and chain elongation in long-chain fatty acid (LCFA) synthesis, the synthesis of LCFAs, palmitate (16:0), stearate (18:0), arachidate (20:0), behenate (22:0), and lignocerate (24:0), in the epidermis, liver, and spinal cord was determined using deuterated water and mass isotopomer distribution analysis in hairless mice and Sprague-Dawley rats. Animals were given 4% deuterated water for 5 days or 8 wk in their drinking water. Blood was withdrawn at the end of these times for the determination of deuterium enrichment, and the animals were killed to isolate the various tissues for lipid extraction for the determination of the mass isotopomer distributions. The mass isotopomer distributions in LCFA were incompatible with synthesis from a single pool of primer. The synthesis of palmitate, stearate, arachidate, behenate, and lignocerate followed the expected biochemical pathways for the synthesis of LCFAs. On average, three deuterium atoms were incorporated for every addition of an acetyl unit. The isotopomer distribution resulting from chain elongation and de novo synthesis can be described by the linear combination of two binomial distributions. The proportions of preexisting, chain elongation, and de novo-synthesized fatty acids as a percentage of the total fatty acids were determined using multiple linear regression analysis. Fractional synthesis was found to vary, depending on the tissue type and the fatty acid, from 47 to 87%. A substantial fraction (24-40%) of the newly synthesized molecules was derived from chain elongation of unlabeled (recycled) palmitate.

The composition and metabolism of fatty acids in Ips paraconfusus Lanier (Coleoptera: Scolytidae)

1972 ◽  
Vol 50 (10) ◽  
pp. 1263-1267 ◽  
Author(s):  
K. R. Penner ◽  
J. S. Barlow
Keyword(s):  
Fatty Acids ◽  
Fatty Acid Composition ◽  
Fatty Acid ◽  
Sexual Dimorphism ◽  
Acid Composition ◽  
De Novo ◽  
Monounsaturated Fatty Acids ◽  
Ips Paraconfusus ◽  
Chain Elongation ◽  
De Novo Synthesis

The fatty acid composition of newly emerged Ips paraconfusus Lanier shows no sexual dimorphism and is approximately as follows: C14:0, 0.5%; C16:0, 23.0%; C16:1, 6%; C18:0, 3%; C18:1, 55%; C18:2, 9%; C18:3, 2%. Both sexes, but particularly the female, use up fatty acids, particularly the monounsaturated acids, during reproduction. Isotope from 1-14C-acetate injected into newly emerged females appeared in all saturated and monounsaturated fatty acids within 30 min. There was evidence of de novo synthesis of C14:0 and C16:0, chain elongation of C16:0 to C18:0, and desaturation of C16:0 and C18:0 to yield C16:1 and C18:1 respectively.

Lipid composition of developing Xenopus laevis embryos

1976 ◽  
Vol 54 (6) ◽  
pp. 578-582 ◽  
Author(s):  
Mary Mes-Hartree ◽  
John B. Armstrong
Keyword(s):  
Fatty Acids ◽  
Fatty Acid Composition ◽  
Fatty Acid ◽  
Xenopus Laevis ◽  
Lipid Content ◽  
Lipid Composition ◽  
De Novo ◽  
Total Lipid Content ◽  
De Novo Synthesis ◽  
Xenopus Laevis Embryos

The total lipid content, amount of phospholipid, proportions of major polar and neutral lipid classes, and the overall fatty acid composition were examined in Xenopus laevis embryos. No obvious differences were observed in any of the parameters between fertilization and hatching, or between eggs produced by different females. The average lipid content per egg was 113 μg, 31.6 μg of which was phospholipid. The major phospholipids were phosphatidylcholine and sphingomyelin. The major fatty acids were palmitic and oleic acids, but polyunsaturated fatty acids were also present in substantial amounts. The results suggest that significant de novo synthesis of lipids does not occur until after hatching.

scholarly journals A Review of the Metabolic Origins of Milk Fatty Acids

2013 ◽  
Vol 5 (3) ◽  
pp. 270-274 ◽  
Author(s):  
Anamaria COZMA ◽  
Doina MIERE ◽  
Lorena FILIP ◽  
Sanda ANDREI ◽  
Roxana BANC ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Mammary Gland ◽  
Fatty Acid ◽  
Milk Fat ◽  
De Novo ◽  
Long Chain Fatty Acids ◽  
De Novo Synthesis ◽  
Ruminal Biohydrogenation ◽  
Long Chain ◽  
Chain Fatty Acids

Milk fat and its fatty acid profile are important determinants of the technological, sensorial, and nutritional properties of milk and dairy products. The two major processes contributing to the presence of fatty acids in ruminant milk are the mammary lipogenesis and the lipid metabolism in the rumen. Among fatty acids, 4:0 to 12:0, almost all 14:0 and about a half of 16:0 in milk fat derive from de novo synthesis within the mammary gland. De novo synthesis utilizes as precursors acetate and butyrate produced through carbohydrates ruminal fermentation and involves acetyl-CoA carboxylase and fatty acid synthetase as key enzymes. The rest of 16:0 and all of the long-chain fatty acids derive from mammary uptake of circulating lipoproteins and nonesterified fatty acids that originate from digestive absorption of lipids and body fat mobilization. Further, long-chain fatty acids as well as medium-chain fatty acids entering the mammary gland can be desaturated via Δ-9 desaturase, an enzyme that acts by adding a cis-9-double bond on the fatty acid chain. Moreover, ruminal biohydrogenation of dietary unsaturated fatty acids results in the formation of numerous fatty acids available for incorporation into milk fat. Ruminal biohydrogenation is performed by rumen microbial population as a means of protection against the toxic effects of polyunsaturated fatty acids. Within the rumen microorganisms, bacteria are principally responsible for ruminal biohydrogenation when compared to protozoa and anaerobic fungi.

scholarly journals Triacylglycerol synthesis in goat mammary gland. The effect of ATP, Mg2+ and glycerol 3-phosphate on the esterification of fatty acids synthesized de novo

1984 ◽  
Vol 220 (2) ◽  
pp. 513-519 ◽  
Author(s):  
H O Hansen ◽  
I Grunnet ◽  
J Knudsen
Keyword(s):  
Fatty Acids ◽  
Mammary Gland ◽  
Fatty Acid ◽  
De Novo ◽  
Fatty Acid Synthetase ◽  
Acid Synthesis ◽  
Medium Chain ◽  
Medium Chain Fatty Acids ◽  
Triacylglycerol Synthesis ◽  
Chain Fatty Acids

Goat mammary-gland microsomal fraction by itself induces synthesis of medium-chain-length fatty acids by goat mammary fatty acid synthetase and incorporates short- and medium-chain fatty acids into triacylglycerol. Addition of ATP in the absence or presence of Mg2+ totally inhibits triacylglycerol synthesis from short- and medium-chain fatty acids, and severely inhibits synthesis de novo of medium-chain fatty acids. The inhibition by ATP of fatty acid synthesis and triacylglycerol synthesis de novo can be relieved by glycerol 3-phosphate. The effect of ATP could not be mimicked by the non-hydrolysable ATP analogue, adenosine 5′-[beta, gamma-methylene]triphosphate and could not be shown to be caused by inhibition of the diacylglycerol acyltransferase by a phosphorylation reaction. Possible explanations for the mechanism of the inhibition by ATP are discussed, and a hypothetical model for its action is outlined.

scholarly journals Inhibition of Very-Long-Chain Fatty Acid Formation by Indanofan, 2-[2-(3-Chlorophenyl)oxiran-2-ylmethyl]-2-ethylindan-1,3-dione, and Its Relatives

2002 ◽  
Vol 57 (1-2) ◽  
pp. 72-74 ◽  
Author(s):  
Hideomi Takahashi ◽  
Jochen Schmalfuß ◽  
Aiko Ohki ◽  
Akemi Hosokawa ◽  
Akira Tanaka ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
De Novo ◽  
Allium Porrum ◽  
Inhibitory Effects ◽  
Strong Activity ◽  
Oxirane Group ◽  
Malonyl Coa ◽  
High Concentrations ◽  
Long Chain

Indanofan and its analogs inhibited the elongation of stearoyl- or arachidoyl-CoA by [2-14C]-malonyl-CoA in leek microsomes from Allium porrum. Although the precise mode of interaction of indanofan at the molecular level is not completely clarified by the present study, it is concluded that indanofan and analogs act as inhibitor of the elongase enzyme involved in de novo biosynthesis of fatty acids with an alkyl chain longer than C18, called very-long-chain fatty acids (VLCFAs). For a strong inhibition of VLCFA formation chloro substituents at the benzene ring and the oxirane group were necessary. Furthermore, the greenhouse test showed strong activity for indanofan and its analogs, and the scores coincided with cell-free elongation inhibition. The cell-free assay, however, failed to indicate any activity for an analog having a methylene instead of the oxirane group, while both Digitaria ciliaris and Echinochloa oryzicola were killed with 1 kg a.i./ha. This finding cannot be discussed because the applied use rate of 1 kg a.i./ha is too high to allow for a score differentiation. For high concentrations of this compound additional unknown inhibitory effects may be involved besides fatty acid elongation.

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