scholarly journals Transformation of Fatty Acids Catalyzed by Cytochrome P450 Monooxygenase Enzymes of Candida tropicalis

2003 ◽  
Vol 69 (10) ◽  
pp. 5992-5999 ◽  
Author(s):  
William H. Eschenfeldt ◽  
Yeyan Zhang ◽  
Hend Samaha ◽  
Lucy Stols ◽  
L. Dudley Eirich ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Cytochrome P450 ◽  
Oleic Acid ◽  
Candida Tropicalis ◽  
Saturated Fatty Acids ◽  
Dicarboxylic Acids ◽  
Initial Step ◽  
Cytochrome P450s ◽  
Commercial Development ◽  
P450 Monooxygenase

ABSTRACT Candida tropicalis ATCC 20336 can grow on fatty acids or alkanes as its sole source of carbon and energy, but strains blocked in β-oxidation convert these substrates to long-chain α,ω-dicarboxylic acids (diacids), compounds of potential commercial value (Picataggio et al., Biotechnology 10:894-898, 1992). The initial step in the formation of these diacids, which is thought to be rate limiting, is ω-hydroxylation by a cytochrome P450 (CYP) monooxygenase. C. tropicalis ATCC 20336 contains a family of CYP genes, and when ATCC 20336 or its derivatives are exposed to oleic acid (C18:1), two cytochrome P450s, CYP52A13 and CYP52A17, are consistently strongly induced (Craft et al., this issue). To determine the relative activity of each of these enzymes and their contribution to diacid formation, both cytochrome P450s were expressed separately in insect cells in conjunction with the C. tropicalis cytochrome P450 reductase (NCP). Microsomes prepared from these cells were analyzed for their ability to oxidize fatty acids. CYP52A13 preferentially oxidized oleic acid and other unsaturated acids to ω-hydroxy acids. CYP52A17 also oxidized oleic acid efficiently but converted shorter, saturated fatty acids such as myristic acid (C14:0) much more effectively. Both enzymes, in particular CYP52A17, also oxidized ω-hydroxy fatty acids, ultimately generating the α,ω-diacid. Consideration of these different specificities and selectivities will help determine which enzymes to amplify in strains blocked for β-oxidation to enhance the production of dicarboxylic acids. The activity spectrum also identified other potential oxidation targets for commercial development.

scholarly journals Identification and Characterization of the CYP52 Family of Candida tropicalis ATCC 20336, Important for the Conversion of Fatty Acids and Alkanes to α,ω-Dicarboxylic Acids

2003 ◽  
Vol 69 (10) ◽  
pp. 5983-5991 ◽  
Author(s):  
David L. Craft ◽  
Krishna M. Madduri ◽  
Mark Eshoo ◽  
C. Ron Wilson
Keyword(s):  
Fatty Acids ◽  
Cytochrome P450 ◽  
Fatty Acid ◽  
Candida Tropicalis ◽  
Strain Improvement ◽  
Dicarboxylic Acids ◽  
Fold Increase ◽  
Isolation And Characterization ◽  
Unique Genes

ABSTRACT Candida tropicalis ATCC 20336 excretes α,ω-dicarboxylic acids as a by-product when cultured on n-alkanes or fatty acids as the carbon source. Previously, a β-oxidation-blocked derivative of ATCC 20336 was constructed which showed a dramatic increase in the production of dicarboxylic acids. This paper describes the next steps in strain improvement, which were directed toward the isolation and characterization of genes encoding the ω-hydroxylase enzymes catalyzing the first step in the ω-oxidation pathway. Cytochrome P450 monooxygenase (CYP) and the accompanying NADPH cytochrome P450 reductase (NCP) constitute the hydroxylase complex responsible for the first and rate-limiting step of ω-oxidation of n-alkanes and fatty acids. 10 members of the alkane-inducible P450 gene family (CYP52) of C. tropicalis ATCC20336 as well as the accompanying NCP were cloned and sequenced. The 10 CYP genes represent four unique genes with their putative alleles and two unique genes for which no allelic variant was identified. Of the 10 genes, CYP52A13 and CYP52A14 showed the highest levels of mRNA induction, as determined by quantitative competitive reverse transcription-PCR during fermentation with pure oleic fatty acid (27-fold increase), pure octadecane (32-fold increase), and a mixed fatty acid feed, Emersol 267 (54-fold increase). The allelic pair CYP52A17 and CYP52A18 was also induced under all three conditions but to a lesser extent. Moderate induction of CYP52A12 was observed. These results identify the CYP52 and NCP genes as being involved in α,ω-dicarboxylic acid production by C. tropicalis and provide the foundation for biocatalyst improvement.

scholarly journals Effect of nitrogen and potassium fertilization on the production and quality of oil in Jatropha curcas L. under the dry and warm climate conditions of Colombia

2014 ◽  
Vol 32 (2) ◽  
pp. 255-265 ◽  
Author(s):  
Omar Montenegro R. ◽  
Stanislav Magnitskiy ◽  
Martha C. Henao T.
Keyword(s):  
Fatty Acids ◽  
Oleic Acid ◽  
Jatropha Curcas ◽  
Oil Content ◽  
Unsaturated Fatty Acids ◽  
Saturated Fatty Acids ◽  
Fruit Production ◽  
Biodiesel Production ◽  
Oil Composition

This study was conducted to assess fruit and seed yield, oil content and oil composition of Jatropha curcas fertilized with different doses of nitrogen and potassium in Espinal (Tolima, Colombia). The yields ranged from 4,570 to 8,800 kg ha-1 of fruits and from 2,430 to 4,746 kg ha-1 of seeds. These yields showed that the fertilizer dose of 150 kg ha-1 N + 120 kg ha-1K increased fruit production by 92% and seed production by 95%, which represents an increase of about 100% in oil production, which increased from 947 to 1,900 kg ha-1. The total oil content in the seeds ranged from 38.7 to 40.1% (w/w) with a high content of the unsaturated fatty acids oleic (> 47%) and linoleic acid (> 29%). The highest content of oleic acid in the seed oil was from the unfertilized control plants and plants with an application of 100 kg ha-1 of N and 60 kg ha-1 of K, with an average of 48%. The lowest content of oleic acid was registered when a low dose of nitrogen and a high level of potassium were applied at a ratio of 1:2.4 and doses of 50 kg ha-1 N + 120 kg ha-1 K, respectively. Low contents of the saturated fatty acids palmitic (13.4%) and stearic (7.26%) were obtained, making this oil suitable for biodiesel production. The nitrogen was a more important nutrient for the production and quality of oil in J. curcas than potassium under the studied conditions of soil and climate.

scholarly journals Antagonism Between Saturated and Unsaturated Fatty Acids in ROS Mediated Lipotoxicity in Rat Insulin-Producing Cells

2015 ◽  
Vol 36 (3) ◽  
pp. 852-865 ◽  
Author(s):  
Wiebke Gehrmann ◽  
Wiebke Würdemann ◽  
Thomas Plötz ◽  
Anne Jörns ◽  
Sigurd Lenzen ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Oleic Acid ◽  
Palmitic Acid ◽  
Unsaturated Fatty Acids ◽  
Saturated Fatty Acids ◽  
H2o2 Production ◽  
Β Cell ◽  
Specific Expression ◽  
Insulin Producing Cells ◽  
H2o2 Formation

Background/Aims: Elevated levels of non-esterified fatty acids (NEFAs) are under suspicion to mediate β-cell dysfunction and β-cell loss in type 2 diabetes, a phenomenon known as lipotoxicity. Whereas saturated fatty acids show a strong cytotoxic effect upon insulin-producing cells, unsaturated fatty acids are not toxic and can even prevent toxicity. Experimental evidence suggests that oxidative stress mediates lipotoxicity and there is evidence that the subcellular site of ROS formation is the peroxisome. However, the interaction between unsaturated and saturated NEFAs in this process is unclear. Methods: Toxicity of rat insulin-producing cells after NEFA incubation was measured by MTT and caspase assays. NEFA induced H2O2 formation was quantified by organelle specific expression of the H2O2 specific fluorescence sensor protein HyPer. Results: The saturated NEFA palmitic acid had a significant toxic effect on the viability of rat insulin-producing cells. Unsaturated NEFAs with carbon chain lengths >14 showed, irrespective of the number of double bonds, a pronounced protection against palmitic acid induced toxicity. Palmitic acid induced H2O2 formation in the peroxisomes of insulin-producing cells. Oleic acid incubation led to lipid droplet formation, but in contrast to palmitic acid induced neither an ER stress response nor peroxisomal H2O2 generation. Furthermore, oleic acid prevented palmitic acid induced H2O2 production in the peroxisomes. Conclusion: Thus unsaturated NEFAs prevent deleterious hydrogen peroxide generation during peroxisomal β-oxidation of long-chain saturated NEFAs in rat insulin-producing cells.

Effect of increasing stearoyl coenzyme A desaturase mRNA concentrations using forage and concentrate diets on the fatty acid composition of ovine adipose tissue

2002 ◽  
Vol 2002 ◽  
pp. 206-206 ◽  
Author(s):  
Z.C.T.R. Daniel ◽  
R.J. Wynn ◽  
A.M. Salter ◽  
P.J. Buttery
Keyword(s):  
Fatty Acids ◽  
Adipose Tissue ◽  
Fatty Acid Composition ◽  
Oleic Acid ◽  
Fatty Acid ◽  
Stearic Acid ◽  
Acid Composition ◽  
Coenzyme A ◽  
Saturated Fatty Acids ◽  
Mrna Levels

Compared to meat from other animals lamb contains high levels of saturated fat, particularly stearic acid which comprises 18% of the total fatty acids (Enser et al, 1996). This stearic acid can be desaturated in the tissue by stearoyl coenzyme A desaturase (SCD) to produce oleic acid. In sheep SCD is produced from a single gene and the levels of SCD mRNA in the tissue correlate well with oleic acid (Ward et al, 1998, Barber et al, 2000) suggesting that an upregulation of SCD activity may increase the relative proportions of unsaturated and saturated fatty acids and so significantly improve the nutritional quality of sheep meat. Our recent studies have shown that insulin increases SCD mRNA levels and monounsaturated fatty acid synthesis in cultured ovine adipose tissue explants (Daniel et al, 2001). The present study was designed to investigate whether feeding a diet believed to manipulate SCD mRNA concentrations would significantly alter the fatty acid composition of lamb.

Enzymatische Aspekte zur Biosynthese des Blatt-Cutins bei Gasteria verricuosa-Blättern nach Verletzung

1963 ◽  
Vol 18 (1) ◽  
pp. 67-79 ◽  
Author(s):  
Wolfgang Heinen ◽  
Ingeborg V. D. Brand
Keyword(s):  
Fatty Acids ◽  
Oleic Acid ◽  
Fatty Acid ◽  
Linoleic Acid ◽  
Mode Of Action ◽  
Saturated Fatty Acids ◽  
Late Phase ◽  
Acid Oxidase ◽  
Enzymatic Assays ◽  
Microscopic Studies

1. Three fatty acid oxidizing enzymes, stearic and oleic acid oxidase as well as lipoxidase have been shown to be present in leaves of Gasteria verricuosa.2. By following the activity of these enzymes after injury we considered that they are involved in cutin synthesis which takes place at the wounded top of the leaf.3. Comparing the activity near the wounded part and the untreated inner sphere of the leaf lead to the conclusion that two of the oxidases (stearic and oleic oxidase) serve as substrate donors for lipoxydase by converting stearic into oleic and the latter into linoleic acid.4. Since the level of polyenic acids in leaves is high in comparison to saturated fatty acids, the activity of stearic and oleic oxidase only increases in the late phase of cutin synthesis, while lipoxydase is highly activated at the top directly after wounding and in the inner part of the leaf 3 - 4 weeks after cutin synthesis has started. At the same time pectinase shows its highest activity, suggesting that the formation of the pectic layer is secondary to the formation of cutin.5. Simultaneously to the enzymatic assays, cutin formation was followed by macro- and microscopic studies.6. The mode of action of lipoxydase and the interrelationship of the oxidizing enzymes in the formation of cutin are discussed and a formula for the structure of Gasteria cutin is given.7. According to the data presented here and the results obtained from literature, a possible scheme for cutin synthesis is given.

scholarly journals Ova Fatty Acids Composition and Spawning Performances of Silver Carp, Hypophthalmichthys molitrix (Morocco)

2020 ◽  
Vol 20 (12) ◽  
pp. 879-888
Author(s):  
Fatima-Zahra Majdoubi ◽  
Redouane Benhima ◽  
Anouar Ouizgane ◽  
Sana Farid ◽  
Mohammed Droussi ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Oleic Acid ◽  
Docosahexaenoic Acid ◽  
Eicosapentaenoic Acid ◽  
Saturated Fatty Acids ◽  
Silver Carp ◽  
Fertilization Success ◽  
Hypophthalmichthys Molitrix ◽  
Maternal Weight ◽  
Significant Difference

The present study is performed in order to determine the relationship between fatty acid (FA) profile of silver carp (Hypophthalmichthys molitrix) eggs and fertilization success, embryos viability and larval production. Mature unfertilized ova were collected during the reproductive season from 23 mature and healthy females, reared in Deroua fish farm (Morocco). Total lipids were extracted from ova sample and subjected to trans-esterification then the resulting fatty acids methyl esters were analyzed by gas chromatography tandem mass spectrometry (GC/MS/MS). The results showed that silver carp ova are dominated on total FA by docosahexaenoic acid (DHA) (21.21%) followed by oleic acid (21.07%), palmitic acid (17.71%) and eicosapentaenoic acid (EPA) (10.25%). During the breeding season, polyunsaturateds (PUFAs), monounsaturateds (MUFAs) and saturated fatty acids (SFA) didn’t show any significant difference. Moreover, within the PUFAs, the n-3 series were more abundant than the n-6 series, the total mean was 31.57 ± 1.01% and 5.33 ± 0.32%, respectively. No correlation was between fatty acids and the fertilization success. Maternal weight has effect on the levels of oleic acid (C18:1), arachidonic acid (C20:4) and docosahexaenoic acid (C22:6) in the egg whereas levels of palmetoleic acid (C16:1), eicosapentaenoic acid (C20:5) and docosahexaenoic acid (C22:6) are related to female age.

scholarly journals Structural insights into oxidation of medium-chain fatty acids and flavanone by myxobacterial cytochrome P450 CYP267B1

2018 ◽  
Vol 475 (17) ◽  
pp. 2801-2817 ◽  
Author(s):  
Ilona K. Jóźwik ◽  
Martin Litzenburger ◽  
Yogan Khatri ◽  
Alexander Schifrin ◽  
Marco Girhard ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Cytochrome P450 ◽  
Saturated Fatty Acids ◽  
Decanoic Acid ◽  
Dodecanoic Acid ◽  
Binding Modes ◽  
Cytochrome P450 Enzymes ◽  
Tetradecanoic Acid ◽  
Medium Chain ◽  
Pharmaceutical Industries

Oxidative biocatalytic reactions performed by cytochrome P450 enzymes (P450s) are of high interest for the chemical and pharmaceutical industries. CYP267B1 is a P450 enzyme from myxobacterium Sorangium cellulosum So ce56 displaying a broad substrate scope. In this work, a search for new substrates was performed, combined with product characterization and a structural analysis of substrate-bound complexes using X-ray crystallography and computational docking. The results demonstrate the ability of CYP267B1 to perform in-chain hydroxylations of medium-chain saturated fatty acids (decanoic acid, dodecanoic acid and tetradecanoic acid) and a regioselective hydroxylation of flavanone. The fatty acids are mono-hydroxylated at different in-chain positions, with decanoic acid displaying the highest regioselectivity towards ω-3 hydroxylation. Flavanone is preferably oxidized to 3-hydroxyflavanone. High-resolution crystal structures of CYP267B1 revealed a very spacious active site pocket, similarly to other P450s capable of converting macrocyclic compounds. The pocket becomes more constricted near to the heme and is closed off from solvent by residues of the F and G helices and the B–C loop. The crystal structure of the tetradecanoic acid-bound complex displays the fatty acid bound near to the heme, but in a nonproductive conformation. Molecular docking allowed modeling of the productive binding modes for the four investigated fatty acids and flavanone, as well as of two substrates identified in a previous study (diclofenac and ibuprofen), explaining the observed product profiles. The obtained structures of CYP267B1 thus serve as a valuable prediction tool for substrate hydroxylations by this highly versatile enzyme and will encourage future selectivity changes by rational protein engineering.

scholarly journals Beneficial effect of oleoylated lipids on paraoxonase 1: protection against oxidative inactivation and stabilization

2003 ◽  
Vol 375 (2) ◽  
pp. 275-285 ◽  
Author(s):  
S. Duy NGUYEN ◽  
Dai-Eun SOK
Keyword(s):  
Fatty Acids ◽  
Oleic Acid ◽  
Linoleic Acid ◽  
Competitive Inhibition ◽  
Protective Action ◽  
Saturated Fatty Acids ◽  
Density Lipoprotein ◽  
Paraoxonase 1 ◽  
Pon1 Activity ◽  
Oxidative Inactivation

The effect of lipids on PON1 (paraoxonase 1), one of antioxidant proteins in high-density lipoprotein, was investigated in respect to inhibition, protection against oxidative inactivation, and stabilization. When the effect of lipids on the PON1 activity was examined, a remarkable inhibition was expressed by polyenoic fatty acids (C18:2–C20:5). Linoleic acid, the most potent (Ki, 3.8 μM), showed competitive inhibition. Next, various lipids were examined for prevention against the inactivation of PON1 by ascorbate/Cu2+, which caused a remarkable (≥90%) inactivation of PON1. Compared with saturated fatty acids (C6–C18), exhibiting a modest protection (9–40%), monoenoic acids (C16:1–C20:1) showed a greater maximal protective effect (Emax, 70–82%), with oleic acid being the most effective (EC50, 2.7 μM). In contrast, polyenoic acids showed no protection. Noteworthy, linoleic acid prohibited the protective action of oleic acid non-competitively. In the structure–activity relationship, a negatively charged group seems to be required for the protective action. Consistent with this, dioleoylphosphatidylglycerol, negatively charged, was more protective than dioleoylphosphatidylcholine. These data, together with requirement of Ca2+ (EC50, 0.6 μM) for the protective action, may support the existence of a specific site responsible for the protective action. A similar protective action of lipids was also observed in the inactivation of PON1 by ascorbate/Fe2+, peroxides or p-hydroxymercuribenzoate. Separately, PON1 was stabilized by oleic acid or oleoylated phospholipids, in combination with Ca2+, but not linoleic acid. These results suggest that in contrast to an adverse action of linoleic acid, monoenoic acids or their phospholipid derivatives play a beneficial role in protecting PON1 from oxidative inactivation as well as in stabilizing PON1.

THE EFFECT OF RAPESEED OIL ON REPRODUCTION AND ON THE COMPOSITION OF RAT MILK FAT

1961 ◽  
Vol 39 (2) ◽  
pp. 195-201 ◽  
Author(s):  
Joyce L. Beare ◽  
E. R. W. Gregory ◽  
D. Morison Smith ◽  
J. A. Campbell
Keyword(s):  
Fatty Acids ◽  
Oleic Acid ◽  
Olive Oil ◽  
Dietary Fat ◽  
Milk Fat ◽  
Rapeseed Oil ◽  
Corn Oil ◽  
Saturated Fatty Acids ◽  
Low Fat ◽  
Low Fat Diet

Rats fed corn oil or a mixture of lard and olive oil produced as many offspring as those receiving no fat supplement with a low-fat commercial meal, but the weanling weight was lower. Although rats fed rapeseed oil continued to reproduce they had fewer and smaller offspring than rats fed other diets.The composition of fatty acids in the milk varied with the dietary fat of the mother. Animals receiving the low-fat diet secreted predominantly saturated fatty acids. A high proportion of linoleic acid appeared in the milk when corn oil was fed, and of oleic acid when the mixture of lard and olive oil was fed. Eicosenoic and erucic acids were present in the milk of rats receiving rapeseed oil, but were less prevalent than in the original oil.

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