• Title/Summary/Keyword: hydroxy fatty acid

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Acetoacetyl-CoA Synthetase, a Novel Cytosolic Ketone Body-Utilizing Enzyme that Specifically Activates Acetoacetate to its Coenzyme A Ester

  • Fukui, Tetsuya
    • Proceedings of the PSK Conference
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    • 2003.10a
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    • pp.70-70
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    • 2003
  • In mammalians, ketone bodies (acetoacetate, D(-)-3-hydroxybutyrate and acetone) are generated mainly in the liver via the 3-hydroxy-3-methylglutaryl-CoA pathway, carried to and utilized in extrahepatic tissues as an energy source during starvation and diabetes in particular due to their overproduction as the consequence of elevated fatty acid oxidation and lowered glucose metabolism. (omitted)

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Study on hydroxy fatty acid contents changes and physiological responses under abiotic stresses in transgenic Camelina

  • Kim, Hyun-Sung;Lee, Hyun-Sook;Lim, Hyun-Gyu;Park, Won;Kim, Hyun-Uk;Lee, Kyeong-Ryeol;Ahn, Sung-Ju
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.191-191
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    • 2017
  • Hydroxy fatty acid (HFA) is an important industrial resource that known to be extracted from seeds of Castor or Lesquerella. However, mass production of HFA from those crops are difficult because of their behavior or life cycle. In this study, we applied HFA synthesis related gene FAH12, RcPDAT1, RcLPCAT, RcDGAT2, and RcPDCT on bioenergy crop Camelina sativa. Furthermore, we determined NaCl or cold stress tolerance changes of transgenic Camelina. RcFAH12, RcPDAT1, RcLPCAT, RcDGAT2, and RcPDCT genes were cloned into multigene expression vector which is engineered with seed specific promoter of FAE1 or Napin. Combination of HFA genes multi-expression vector constructs were divided into Set3 (RcFAH12, RcPDAT1-2, RcLPCAT), Set4 (RcFAH12, RcDGAT2, RCPDAT1-2, RcLPCAT), and Set5 (RcFAH12, RcDGAT2, RCPDAT1-2, RcLPCAT, RcPDCT). Transgenic HFA synthesis Camelina plants were generated using agrobacterium-mediated vacuum infiltration system. Results of fatty acid composition of T1 transgenic Camelina seeds analyzed by GC-MS showed 9.5, 9.0, and 13.6 % of HFA contents in Set3#6, Set4#8, and Set5#10, respectively. Therefore, seeds of T2 generation were harvest from Set5#10 which is shown highest HFA contents, and, 17.7, 8.1 and 10.5 % of HFA contents were determined in Set5#10-5, Set5#10-8, and Set#10-10, respectively. However, 7.7% of C18:2 and 22.3 % of C18:3 among unsaturated fatty acids were decreased in Set5#10-5 than WT. Meanwhile, we confirmed abiotic stress responses in T2 transgenic Camelina Set5#10-5 and Set5#10-10 under 0, 100, 150, and 200 mM NaCl or 25, 15, and $10^{\circ}C$ temperature for 5 weeks. Both Set5#10-5 and Set5#10-10 showed lower growth in height than WT in control and NaCl condition. Growth of leaf length and width were similar in WT and Set5#10-10 but lower in Set5#10-5 under NaCl stress. Number of opened flowers showed that both transgenic Camelina were lower than WT under normal condition. But, WT and Set5#10-10 showed similar opened flower number in 100 and 200 mM NaCl. In cold stress, 15 and $10^{\circ}C$ treatment for 5 weeks did not showed significant changes in between WT and both transgenic lines even they showed different growth rate in control condition. Taken together, growth and development are delayed by expression of exogenous HFA related genes in transgenic lines but relative abiotic stress sensitivity is similar with WT. In conclusion, reduced C18:2 or C18:3 fatty acid composition of seed by HFA synthesis is resulted from lack of resource supplement for development at seedling stage but it is not affect NaCl and cold stress tolerance.

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In vitro control of plant pathogens by using anti-fungal hydroxy fatty acid obtained from microbial bioconversion of linolenic acid

  • Kang, Sun-Chul;Bajpai, Vivek K.;Kim, Hak-Ryul
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.309-310
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    • 2005
  • Bioconverted linolenic acid (bLNA) obtained from linolenic acid by Pseudomonas aeruginosa PR3, showed anti-fungal activity against plant pathogens such as B. cinerea, F. oxysporum, F. solani, P. capsici and C. capsici. The oil sample also showed anti-fungal activity with MIC values, ranging from >250 to >1,000 ${\mu}g/ml$. Varied concentrations of bLNA had a great potential effect on spore germination of different fungi.

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A Lipopeptide Biosurfactant Produced by Bacillus subtilis C9 Selected through the Oil Film-collapsing Assay

  • Kim, Hee-Sik;Lee, Chang-Ho;Suh, Hyun-Hyo;Ahn, Keug-Hyun;Oh, Hee-Mock;Kwon, Gi-Seok;Yang, Ji-Won;Yoon, Byung-Dae
    • Journal of Microbiology and Biotechnology
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    • v.7 no.3
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    • pp.180-188
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    • 1997
  • Bacillus subtilis C9 was selected by measuring the oil film-collapsing activity and produced biosurfactant in a medium containing glucose as a sole carbon source. The biosurfactant emulsified hydrocarbons, vegetable oils and crude oil, and lowered the surface tension of culture broth to 28 dyne/cm. A biosurfactant, C9-BS produced by B. subtilis C9 was purified by ultrafiltration, extraction with chloroform and methanol, adsorption chromatography, and preparative reversed phase HPLC. Structural analyses, IR spectroscopy, FAB mass spectroscopy, amino acid composition, and NMR analyses, demonstrated that C9-BS was a lipopeptide comprising a fatty acid tail and peptide moiety. The lipophilic part consisting of $C_{14}\;or\;C_{15}$ hydroxy fatty acid was linked to the hydrophilic peptide part, which contained seven amino acids (Glu-Leu-Leu-Val-Asp-Leu-Leu) with a lactone linkage.

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Carrot Juice Administration Decreases Liver Stearoyl-CoA Desaturase 1 and Improves Docosahexaenoic Acid Levels, but Not Steatosis in High Fructose Diet-Fed Weanling Wistar Rats

  • Mahesh, Malleswarapu;Bharathi, Munugala;Reddy, Mooli Raja Gopal;Kumar, Manchiryala Sravan;Putcha, Uday Kumar;Vajreswari, Ayyalasomayajula;Jeyakumar, Shanmugam M.
    • Preventive Nutrition and Food Science
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    • v.21 no.3
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    • pp.171-180
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    • 2016
  • Non-alcoholic fatty liver disease (NAFLD) is one of the most prevalent liver diseases associated with an altered lifestyle, besides genetic factors. The control and management of NAFLD mostly depend on lifestyle modifications, due to the lack of a specific therapeutic approach. In this context, we assessed the effect of carrot juice on the development of high fructose-induced hepatic steatosis. For this purpose, male weanling Wistar rats were divided into 4 groups, fed either a control (Con) or high fructose (HFr) diet of AIN93G composition, with or without carrot juice (CJ) for 8 weeks. At the end of the experimental period, plasma biochemical markers, such as triglycerides, alanine aminotransferase, and ${\beta}$-hydroxy butyrate levels were comparable among the 4 groups. Although, the liver injury marker, aspartate aminotransferase, levels in plasma showed a reduction, hepatic triglycerides levels were not significantly reduced by carrot juice ingestion in the HFr diet-fed rats (HFr-CJ). On the other hand, the key triglyceride synthesis pathway enzyme, hepatic stearoyl-CoA desaturase 1 (SCD1), expression at mRNA level was augmented by carrot juice ingestion, while their protein levels showed a significant reduction, which corroborated with decreased monounsaturated fatty acids (MUFA), particularly palmitoleic (C16:1) and oleic (C18:1) acids. Notably, it also improved the long chain n-3 polyunsaturated fatty acid, docosahexaenoic acid (DHA; C22:6) content of the liver in HFr-CJ. In conclusion, carrot juice ingestion decreased the SCD1-mediated production of MUFA and improved DHA levels in liver, under high fructose diet-fed conditions. However, these changes did not significantly lower the hepatic triglyceride levels.

Determination of plasma ketone body following oximation-trimethylsily| derivatization using gas chromatography-mass spectrometry selected ion monitoring (혈장 중 케톤체의 옥심-TMS 유도체화 후 GC-MS/SIM을 이용한 분석)

  • Yoon, Hye-Ran
    • Analytical Science and Technology
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    • v.29 no.1
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    • pp.49-55
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    • 2016
  • A ketone body (acetoacetic acid, β-hydroxybutyric acid, and acetone) increases from blood or urine when bio-energy dependence pays more fatty acid than glucose. However, in case oxidation of fat is greater than the capacity of the citric acid cycle the fatty acid oxidation is made from acetoacetyl CoA to acetoacetate then, again form β-hydroxyburytic acid to acetone, the diffusion take place into the blood. Enzymes that oxidize ketone body in the brain and nerve tissue blood ketone dody is increased during prolonged fasting, brain used it as energy. In this study, we developed the rapid two step derivatization method for sensitive detection of the ketone body by GC-MS/SIM. The plasma was deproteinized and then the hydroxy and carboxyl groups of ketone body are subjected to extraction and drying then, keto-group were derivatized with hydoxylamine at 60℃ for 30 min for oximation. Then it was trimetyl-silylated with BSTFA at 80℃ for 30 min and analyzed using a GC-MS. The linear ranges were in between 0.001 μg/mL and 250 μg/mL for β-hydroxy butyrate, and acetoacetate. The method detection limits were below 0.1 pg over each target compound determined. The mean recoveries (%) of target compounds were ranged from 88.2 % to 92.3 % at 1 µg/mL, from 89.5 % to 94.8 % at 10 μg/mL, with RSD of 6.3-9.4 %. This method could be applied to quantification of ketone bodies which are seen in the keto-acidosis in children and adults from a variety of diseases that cause ketones in the blood and urine.

Isolation and Characterization of Major Glycosphingolipid from Rice Bran Extract (쌀겨 추출물로부터 스핑고당지질의 분리와 구조결정)

  • Mitsutake, Susumu;Okada, Tadashi;Kang, Byoung-Won
    • Applied Biological Chemistry
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    • v.50 no.1
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    • pp.72-76
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    • 2007
  • In order to examine the biofunctions of glycosylceramide which is representative of sphingolipid, monoglycosylceramide (cerebroside) was isolated from rice bran extract. Crude glycosylceramides were isolated in large quantities and promptly by flash system column chromatography from rice bran extract, and purified by normal-phase HPLC using an evaporative light-scattering detector. One major cerebroside was obtained by HPLC used as an eluent consisting of chloroform, methanol and water (99:11:1, v/v/v), and the constituents were analyzed by MALDI/TOF-MS, FAB-MS, GC and 600 MHz $^1$H-NMR. The component sugar was estimated to be glucose. In the ceramide, the fatty acid component consist was 2-hydroxy arachidic acid. The long-chain base component was sphinga-4,8-dienine.

Biological Control of Phytophthora Blight of Red-pepper Caused by Phytophthora capsici.;Ⅲ. Identification of the Antifungal Substances Produced by Pseudomonas sp. A - 183. (고추역병균(疫病菌)(Phytophthora capsici)의 생물학적(生物學的) 방제(防除);Ⅲ. 항균물질(抗菌物質)의 구조분석(構造分析))

  • Chang, Yoon-Hee;Jang, Sang-Moon;Choi, Jyung;Lee, Dong-Hoon
    • Korean Journal of Environmental Agriculture
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    • v.16 no.1
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    • pp.1-6
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    • 1997
  • This study was carried out to identify the three antifungal substances isolated from the culture medium of Pseudomonas sp. A-183 which is antagonistic against Phytophthora capsici. The substance A and B showed positive reactions at the Molish test and Anthrone test, but negative one at the Fehling test, strongly suggesting that both substance A and B had nonreducing sugar frameworks. The substance C only exhibited the phenomenon of the UV induced fluorescence. From the qualitative analysis with the spectroscopic techniques such as UV, Mass, IR and NMR, the substance A and B were known to be composed to sugar and fatty acid, and showed a base peak of 171(m/e). It was identified that substance A was $(2-O-L-rhamnosyl-{\alpha}-L-rhamnosyl-{\beta}-hydroxydecanoyl-{\beta}-hydroxy$ decanoic acid) and the substance B was $({\alpha}-L-rhamnosyl-{\beta}-hydroxydecanoyl-{\beta}-hydroxy$ decanoic acid). The substance C was identified as a phenazine from the results of qualitative analysis with the spectroscopic techniques such as UV, Mass, IR and NMR.

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Functional Characterization of Drosophila melanogaster CYP6A8 Fatty Acid Hydroxylase

  • Sang-A Lee;Vitchan Kim;Byoungyun Choi;Hyein Lee;Young-Jin Chun;Kyoung Sang Cho;Donghak Kim
    • Biomolecules & Therapeutics
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    • v.31 no.1
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    • pp.82-88
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    • 2023
  • Genomic analysis indicated that the genome of Drosophila melanogaster contains more than 80 cytochrome P450 genes. To date, the enzymatic activity of these P450s has not been extensively studied. Here, the biochemical properties of CYP6A8 were characterized. CYP6A8 was cloned into the pCW vector, and its recombinant enzyme was expressed in Escherichia coli and purified using Ni2+-nitrilotriacetate affinity chromatography. Its expression level was approximately 130 nmol per liter of culture. Purified CYP6A8 exhibited a low-spin state in the absolute spectra of the ferric forms. Binding titration analysis indicated that lauric acid and capric acid produced type I spectral changes, with Kd values 28 ± 4 and 144 ± 20 µM, respectively. Ultra-performance liquid chromatography-mass spectrometry analysis showed that the oxidation reaction of lauric acid produced (ω-1)-hydroxylated lauric acid as a major product and ω-hydroxy-lauric acid as a minor product. Steady-state kinetic analysis of lauric acid hydroxylation yielded a kcat value of 0.038 ± 0.002 min-1 and a Km value of 10 ± 2 µM. In addition, capric acid hydroxylation of CYP6A8 yielded kinetic parameters with a kcat value of 0.135 ± 0.007 min-1 and a Km value of 21 ± 4 µM. Because of the importance of various lipids as carbon sources, the metabolic analysis of fatty acids using CYP6A8 in this study can provide an understanding of the biochemical roles of P450 enzymes in many insects, including Drosophila melanogaster.

Synthesis and Characterization of Lithium Dual Complex Grease (Lithium Dual Complex 그리이스의 합성 및 특성연구)

  • 최웅수;권오관;문탁진;유영홍
    • Tribology and Lubricants
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    • v.1 no.1
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    • pp.80-87
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    • 1985
  • Lithium dual complex grease was prepared through the second continuous saponification reaction of a complex gellant system whose essential components comprised of a selected hydroxy fatty acid, lithium hydroxide monohydrate and boric acid to have a fiber structure of chemical, thermal and mechanical stability at high temperatures. An optimum amount of complex gellant was found to be 14% (NLGI #2), and an addition of castor wax of 1.5% provided an excellent performance properties, especially. The oil separation, oxidation stability, water wash-out property, shear stability, extreme pressure and wear property of thus prepared were tested by the ASTM and KS methods, and a characteristic result was obtained.