• Title/Summary/Keyword: Long Chain Fatty Acids

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Molecular Cloning, Characterization, and Expression Analysis of Chicken Δ-6 Desaturase

  • Kang, Xiangtao;Bai, Yichun;Sun, Guirong;Huang, Yanqun;Chen, Qixin;Han, Ruili;Li, Guoxi;Li, Fadi
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.1
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    • pp.116-121
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    • 2010
  • Long-chain polyunsaturated fatty acids (LC-PUFA) promote the development of brain and vision of the fetus, relieve inflammation, inhibit oral dysplasia of rumor cell, decrease the incidence of cardiovascular disease and regulate arrhythmia. ${\Delta}-6$ desaturase is the rate-limited enzyme in the desaturation process. This study reports the cloning, characterization and tissue expression of a ${\Delta}-6$ desaturase gene in the chicken. PCR primers were designed based on the predicted sequence of chicken ${\Delta}-6$ desaturase (accession number: XM421053) and used to isolate a cDNA fragment of 1,323 bp from chicken liver. Based on the 1,323 bp fragment an EST (BI390105) was obtained by BLAST. The EST and 5'nd of the 1,323 bp fragment were partially overlapped. Gene specific primers derived from the EST were used for amplification of the 5'nd. Another gene-specific primer derived from the 1,323 bp fragment was used for amplification of the 3'nd by 3'ACE. Then the three overlapping cDNA sequences obtained were assembled with DNAMAN software and a full-length ${\Delta}-6$ desaturase of 2,153 bp was obtained. The full-length cDNA contained an ORF of 1,335 bp with a 5'ntranslated region of 147 nucleotides followed by an ATG initiation codon. Stop codon TGA was at position 1,481-1,483 bp. The deduced amino acids shared an homology above 77% with bovine, mice, orangutan, rat and human. The protein sequence had three histidine-rich regions HDFGH (HisI region), HFQHH (HisII region) and HH (HisIII region), a cytochrome $b_{5}$-like domain containing a heme-binding motif and two transmembrane domains. Sequence analysis of the chicken genomic DNA revealed that the coding sequence of chicken ${\Delta}-6$ desaturase included 12 exons and 11 introns. Semi-quantitative RT-PCR showed that the ${\Delta}-6$ desaturase expression levels were in turn liver, spleen, pancreas, lung, breast muscle, heart, and abdominal fat. The expression of ${\Delta}-6$ desaturase in liver was significantly higher than that in breast muscle (p<0.01). The expression of ${\Delta}-6$ desaturase in lung was significantly higher than that in abdominal fat (p<0.01). This is the first clone of chicken ${\Delta}-6$ desaturase.

Addition Effect of Seed-associated or Free Linseed Oil on the Formation of cis-9, trans-11 Conjugated Linoleic Acid and Octadecenoic Acid by Ruminal Bacteria In Vitro

  • Wang, J.H.;Song, M.K.;Son, Y.S.;Chang, M.B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.8
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    • pp.1115-1120
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    • 2002
  • The effects of seed-associated or free linseed oil on fermentation characteristics and long-chain unsaturated fatty acids composition, especially the formation of conjugated linoleic acid (CLA) and octadecenoic acid (trans-11 $C_{18:1}$, $t-C_{18:1}$) by mixed ruminal bacteria were examined in vitro. Concentrate (1% of culture solution, w/v, as-fed basis) with ground linseed (0.6% of culture solution, w/v, DM basis) or linseed oil as absorbed onto ground alfalfa hay was added to 600 ml mixed solution consisting of strained rumen fluid and artificial saliva at the ratio of 1:1 in a glass culture jar. The culture jar was covered with a glass lid with stirrer, and placed into a water-bath ($39^{\circ}C$) and incubated anaerobically up to 24 h. Seed-associated or free linseed oil did not significantly affect the pH and ammonia concentration in the culture solution. Molar percent of acetate tended to increase while that of propionate decreased with the addition of free oil treatment throughout the incubation. Differences in bacterial number were relatively small, regardless of the form of supplements. Decreasing trends in the compositions of linoleic acid ($C_{18:2}$) and linolenic acid ($C_{18:3}$) but increasing trends of stearic acid ($C_{18:0}$), $t-C_{18:1}$ and CLA compositions were found from culture contents up to 12h incubation when incubated with both ground linseed and linseed oil. The compositions of $C_{18:0}$, $C_{18:2}$ and $C_{18:3}$ were greater but those of oleic acid ($C_{18:1}$), $t-C_{18:1}$ and CLA were smaller in a culture solution containing ground linseed than those containing linseed oil. The ratio of $t-C_{18:1}$ to CLA was lower in the culture solutions containing linseed oil up to 12h incubations as compared to those containing ground linseed.

Effect of Continuous use of Inorganic Fertilizer on the Soil Organisms and Food Chain (무기질비료의 장기연용이 토양생물 및 먹이연쇄에 미치는 영향)

  • Eo, Jinu;Park, Kee-Choon;Park, Jin-Myeon;Kim, Myung-Hyun;Choi, Soon-Kun;Bang, Hea-Seon
    • Korean Journal of Environmental Agriculture
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    • v.35 no.1
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    • pp.39-45
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    • 2016
  • BACKGROUND: This study aimed to evaluate the combined effects of three components (NPK) of chemical fertilizers with basal application of compost on soil organisms.METHODS AND RESULTS: The soil was treated with five treatments continuously for 15 years: control, PK, NK, NP and NPK. The application of N increased plant growth or biomass, and enhanced organic matter content in the soils. Levels of microbial phospholipid fatty acids (PLFAs) in the soils did not show marked differences among the soils treated with different treatments. However, the principal component analysis showed the changes in the structure of the microbial community in the soil, depending on treatments added. Nitrogen application caused a decrease of pH and an increase of EC in the soils, and these environmental stresses appeared to offset the promoting effect of increased organic matter content on microbial abundance. The abundance of bacterivorous nematodes was the highest in the soils after treating NPK; however, the abundance of fungivorous nematodes was unaffected. There was no significant correlation between the abundances of microbial groups and their feeders. Organic matter content was significantly correlated with the abundance of nematodes in the soils.CONCLUSION: Our results showed that chemical fertilizers affect the soil food chains through both biotic and abiotic factors, and a trophic cascade in the soils may not occur in response to long-term fertilization.

Production of Biodiesel Using Immobilized Lipase from Proteus vulgaris (Proteus vulgaris에서 유래한 리파아제의 고정화 및 바이오디젤 생산)

  • Yoon, Shin-Ah;Han, Jin-Yee;Kim, Hyung-Kwoun
    • Microbiology and Biotechnology Letters
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    • v.39 no.3
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    • pp.238-244
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    • 2011
  • Biodiesel, mono-alkyl esters of long chain fatty acids, is one of the alternative fuels derived from renewable lipid feedstock, such as vegetable oils or animal fats. For decade, various lipases have been used for the production of biodiesel. However, the production of biodiesel by enzymatic catalyst has profound restriction in industry application due to high cost. To overcome these problems, many research groups have studied extensively on the selection of cheap oil sources, the screening of suitable lipases, and development of lipase immobilization methods. In this study, we produced biodiesel from plant oil using Proteus vulgaris lipase K80 expressed in Escherichia coli cells. The recombinant lipase K80 was not only expressed in high level but also had high specific lipase activity and high stability in various organic solvents. Lipase K80 could produce biodiesel from olive oil by 3-stepwise methanol feeding method. The immobilized lipase K80 also produced biodiesel using the same 3-stepwise method. The immobilized lipase could produce biodiesel efficiently from various plant oils and waste oils.

A Study on The Preparation of Poly(alkyl methacrylate-co-maleic anhydride) as Cold Flow Improvers for Biodiesel Fuels (바이오디젤용 저온 유동성 향상제로서의 폴리 (알킬메타크릴레이트-공-무수말레인산) 제조 연구)

  • Hong, Jin-Sook;Chung, Keun-Wo;Kim, Young-Wun;Kim, Nam-Kyun;Im, Dae-Jae
    • Applied Chemistry for Engineering
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    • v.23 no.2
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    • pp.232-240
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    • 2012
  • Bio-diesel (BD) is the mono alkyl esters of long chain fatty acids derived from renewable feed stocks like vegetable oils or animal fats. Bio-diesel shows poorer fuel properties than that of diesel fuel in a cold condition. For the diesel fuel, many cold flow improvers have been developed; however, since primary ingredients of bio-diesel are different from those of the diesel fuel, there is a limit to the cold flow improvement when the same cold flow improvers are added to bio diesel. In this study, to improve low temperature properties of bio-diesel, we developed a cold flow improver using an alkyl methacrylate monomer, prepared via ester reaction, and maleic anhydride and also conducted a ring opening reaction using amine. We characterized the products using $^1H-NMR$, FT-IR and GPC methods. In addition, the cold flow improvements of the products in Soybean BD and Palm BD in the concentration rage of 1000~10000 ppm were investigated. It was found that the addition of LMA2SMA6MA2-C8A in Soybean BD improved the pour point by $12.5\;^{\circ}C$.

Available phosphorus levels modulate gene expression related to intestinal calcium and phosphorus absorption and bone parameters differently in gilts and barrows

  • Julia Christiane Votterl;Jutamat Klinsoda;Simone Koger;Isabel Hennig-Pauka;Doris Verhovsek;Barbara U. Metzler-Zebeli
    • Animal Bioscience
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    • v.36 no.5
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    • pp.740-752
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    • 2023
  • Objective: Dietary phytase increases bioavailability of phytate-bound phosphorus (P) in pig nutrition affecting dietary calcium (Ca) to P ratio, intestinal uptake, and systemic utilization of both minerals, which may contribute to improper bone mineralization. We used phytase to assess long-term effects of two dietary available P (aP) levels using a one-phase feeding system on gene expression related to Ca and P homeostasis along the intestinal tract and in the kidney, short-chain fatty acids in stomach, cecum, and colon, serum, and bone parameters in growing gilts and barrows. Methods: Growing pigs (37.9±6.2 kg) had either free access to a diet without (Con; 75 gilts and 69 barrows) or with phytase (650 phytase units; n = 72/diet) for 56 days. Samples of blood, duodenal, jejunal, ileal, cecal, and colonic mucosa and digesta, kidney, and metacarpal bones were collected from 24 pigs (6 gilts and 6 barrows per diet). Results: Phytase decreased daily feed intake and average daily gain, whereas aP intake increased with phytase versus Con diet (p<0.05). Gilts had higher colonic expression of TRPV5, CDH1, CLDN4, ZO1, and OCLN and renal expression of TRPV5 and SLC34A3 compared to barrows (p<0.05). Phytase increased duodenal expression of TRPV5, TRPV6, CALB1, PMCA1b, CDH1, CLDN4, ZO1, and OCLN compared to Con diet (p<0.05). Furthermore, phytase increased expression of SCL34A2 in cecum and of FGF23 and CLDN4 in colon compared to Con diet (p<0.05). Alongside, phytase decreased gastric propionate, cecal valerate, and colonic caproate versus Con diet (p<0.05). Phytase reduced cortical wall thickness and index of metacarpal bones (p<0.05). Conclusion: Gene expression results suggested an intestinal adaptation to increased dietary aP amount by increasing duodenal trans- and paracellular Ca absorption to balance the systemically available Ca and P levels, whereas no adaption of relevant gene expression in kidney occurred. Greater average daily gain in barrows related to higher feed intake.

Development and Validation of an Official Analytical Method for Determination of Ipfencarbazone in Agricultural Products using GC-ECD (GC-ECD를 이용한 농산물 중 Ipfencarbazone의 신규분석법 개발 및 검증)

  • Jang, Jin;Kim, Heejung;Lee, Eun-Hyang;Ko, Ah-Young;Ju, Yunji;Kim, Sooyeon;Chang, Moon-Ik;Rhee, Gyu-Seek
    • The Korean Journal of Pesticide Science
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    • v.19 no.3
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    • pp.210-217
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    • 2015
  • Ipfencarbazone is a herbicide of the tetrazolinone class, and is believed to be an inhibitor of very long chain fatty acids (VLCFAs), which control cell division in weeds. The objective of this study was to develop and validate an official analytical method for ipfencarbazone determination in agricultural products. The ipfencarbazone residues in agricultural products were extracted with acetone, partitioned with n-hexane, and then purified through silica SPE cartridge. Finally, the analyte was quantified by gas chromatograph-electron capture detector (GC-ECD) and confirmed with gas chromatograph/mass spectrometer(GC/MS). The linear range of ipfencarbazone was 0.01 to 1.0 mg/L with the coefficient of determination ($r^2$) of 0.9999. The limit of detection (LOD) and quantification (LOQ) was 0.003 and 0.01 mg/kg, respectively. In addition, average recoveries of ipfencarbazone ranged from 80.6% to 112.3% at the different concentration levels LOQ, 10LOQ and 50LOQ, while the relative standard deviation was 2.2-8.6%. All values were consistent with the criteria ranges requested in the CODEX guidelines. Furthermore, and inter-laboratory study was conducted to validate the method. This proposed method for determination of ipfencarbazone residues in agricultural products can be used as an official analytical method.