scholarly journals Control mechanisms operating for lipid biosynthesis differ in oil-palm (Elaeis guineensis Jacq.) and olive (Olea europaea L.) callus cultures

2002 ◽  
Vol 364 (2) ◽  
pp. 385-391 ◽  
Author(s):  
Umi S. RAMLI ◽  
Darren S. BAKER ◽  
Patti A. QUANT ◽  
John L. HARWOOD
Keyword(s):  
Fatty Acid ◽  
Oil Palm ◽  
Olea Europaea ◽  
Elaeis Guineensis ◽  
Lipid Synthesis ◽  
Control Analysis ◽  
Flux Control ◽  
Olea Europaea L ◽  
Elaeis Guineensis Jacq ◽  
Olea Europaéa

As a prelude to detailed flux control analysis of lipid synthesis in plants, we have examined the latter in tissue cultures from two important oil crops, olive (Olea europaea L.) and oil palm (Elaeis guineensis Jacq.). Temperature was used to manipulate the overall rate of lipid formation in order to characterize and validate the system to be used for analysis. With [1-14C]acetate as a precursor, an increase in temperature from 20 to 30°C produced nearly a doubling of total lipid labelling. This increase in total lipids did not change the radioactivity in the intermediate acyl-(acyl carrier protein) or acyl-CoA pools, indicating that metabolism of these pools did not exert any significant constraint for overall synthesis. In contrast, there were some differences in the proportional labelling of fatty acids and of lipid classes at the two temperatures. The higher temperature caused a decrease in polyunsaturated fatty acid labelling and an increase in the proportion of triacylglycerol labelling in both calli. The intermediate diacylglycerol was increased in olive, but not in oil palm. Overall the data indicate the suitability of olive and oil-palm cultures for the study of lipid synthesis and indicate that de novo fatty acid synthesis may exert more flux control than complex lipid assembly. In olive, diacylglycerol acyltransferase may exert significant flux control when lipid synthesis is rapid.

Comparative study on fatty acid composition of olive (Olea europaea L.), with emphasis on phytosterol contents

2017 ◽  
Vol 31 (8) ◽  
pp. e3933 ◽  
Author(s):  
Ali Ozkan ◽  
Hassan Y. Aboul-Enein ◽  
Muhittin Kulak ◽  
Recep Bindak
Keyword(s):  
Fatty Acid Composition ◽  
Fatty Acid ◽  
Comparative Study ◽  
Acid Composition ◽  
Olea Europaea ◽  
Olea Europaea L ◽  
Olea Europaéa

scholarly journals Control analysis of lipid biosynthesis in tissue cultures from oil crops shows that flux control is shared between fatty acid synthesis and lipid assembly

2002 ◽  
Vol 364 (2) ◽  
pp. 393-401 ◽  
Author(s):  
Umi S. RAMLI ◽  
Darren S. BAKER ◽  
Patti A. QUANT ◽  
John L. HARWOOD
Keyword(s):  
Fatty Acid ◽  
Fatty Acid Synthesis ◽  
Oil Palm ◽  
Biosynthetic Pathway ◽  
Acid Synthesis ◽  
Lipid Biosynthesis ◽  
Control Analysis ◽  
Tissue Cultures ◽  
Flux Control ◽  
Oil Crops

Top-Down (Metabolic) Control Analysis (TDCA) was used to examine, quantitatively, lipid biosynthesis in tissue cultures from two commercially important oil crops, olive (Olea europaea L.) and oil palm (Elaeis guineensis Jacq.). A conceptually simplified system was defined comprising two blocks of reactions: fatty acid synthesis (Block A) and lipid assembly (Block B), which produced and consumed, respectively, a common and unique system intermediate, cytosolic acyl-CoA. We manipulated the steady-state levels of the system intermediate by adding exogenous oleic acid and, using two independent assays, measured the effect of the addition on the system fluxes (JA and JB). These were the rate of incorporation of radioactivity: (i) through Block A from [1-14C]acetate into fatty acids and (ii) via Block B from [U-14C]glycerol into complex lipids respectively. The data showed that fatty acid formation (Block A) exerted higher control than lipid assembly (Block B) in both tissues with the following group flux control coefficients (C):(i) Oil palm: ∗CJTLBlkB = 0.64±0.05 and ∗CJTLBlkB = 0.36±0.05(ii) Olive: ∗CJTLBlkB =0.57±0.10 and ∗CJTLBlkB = 0.43±0.10where ∗C indicates the group flux control coefficient over the lipid biosynthesis flux (JTL) and the subscripts BlkA and BlkB refer to defined blocks of the system, Block A and Block B. Nevertheless, because both parts of the lipid biosynthetic pathway exert significant flux control, we suggest strongly that manipulation of single enzyme steps will not affect product yield appreciably. The present study represents the first use of TDCA to examine the overall lipid biosynthetic pathway in any tissue, and its findings are of immediate academic and economic relevance to the yield and nutritional quality of oil crops.

scholarly journals Kloning parsial gen penyandi enoil-ACP reduktase dari mesokarp buah kelapa sawit (Elaeis guineensis Jacq.) Partial cloning of gene encoding enoyl-ACP reductase from mesocarp of oil palm (Elaeis guineensis Jacq.)

2016 ◽  
Vol 72 (1) ◽  
Author(s):  
Asmini BUDIANI ◽  
Djoko SANTOSO ◽  
Hajrial ASWIDINNOOR ◽  
Antonius SUWANTO
Keyword(s):  
Fatty Acid ◽  
Oil Palm ◽  
Fatty Acid Synthase ◽  
Elaeis Guineensis ◽  
Rt Pcr ◽  
Gene Encoding ◽  
Oil Accumulation ◽  
Enoyl Acp Reductase ◽  
Conserved Region ◽  
Elaeis Guineensis Jacq

Summary Enoyl-ACP reductase (ENR) is a component of fatty acid synthase (FAS) that is considered to play an important role in fatty acid elongation and oil accumulation of several plants. One of the proteins expressed coinciding with fruit development and oil accumulation in oil palm has been detected from the previous study and had homology with ENR. Therefore, as a part of genetic engineering program to improve oil content and quality in oil palm mesocarp, this research was aimed to clone cDNA conserved region of gene encoding enoyl-ACP reductase from oil palm mesocarp. Based on the amino acid sequence of the polypeptide that was homologous with ENR and combined with information of conserved region sequences of the same gene from other plant sources, primers were designed for amplifying conserved region of the ENR gene. Amplifi-cation was carried out by RT-PCR using total RNA as template, at several annealing temperatures and MgCl2 concentrations. Amplification product was cloned using pCR 2.1-TOPO, and the sequence was subjected into BlastN analysis. The results confirmed that the cloned cDNA fragment with 698 bp in size was the conserved region of the ENR gene.  This sequence was highly homologous with the same gene from other plants such as Oryza sativa, Olea europaea, Brassica napus, Triticum aestivum and Arabidopsis thaliana with E-value 1e-96, 7e-77, 2e-64, 5e-41 and 3e-36, respectively. Based on this result, primers have been made and used to amplify the 5’- and 3’ ends of the ENR -cDNA  of oil palm mesocarp. Ringkasan Enoil-ACP reduktase (ENR) merupakan salah satu komponen asam lemak sintase (FAS) yang berperan penting dalam pemanjangan asam lemak dan akumulasi minyak pada berbagai tanaman. Salah satu protein yang ter-ekspresi sejalan dengan tahapan perkembangan buah sawit dan akumulasi minyak pada penelitian sebelumnya diketahui mempunyai homologi dengan ENR. Oleh karena itu, sebagai salah satu bagian dari usaha rekayasa metabolisme minyak pada mesokarp buah sawit, penelitian ini bertujuan untuk mengklon cDNA daerah konservatif gen penyandi ENR dari mesokarp buah sawit. Berdasarkan  sekuen asam amino polipeptida yang mempunyai homologi dengan ENR dan dikombinasikan dengan hasil penjajaran daerah konservatif gen tersebut dari berbagai anaman lain, dirancang primer  untuk  amplifikasi daerah konservatif ENR. Amplifikasi dilakukan dengan RT-PCR menggunakan templat RNA total pada berbagai suhu   penempelan   dan   konsentrasi    MgCl2. Hasil amplifikasi dimurnikan dari gel dan diklon menggunakan vektor kloning pCR2.1-TOPO serta dianalisis nya menggunakan BlastN. Hasilnya mengkonfirmasi fragmen cDNA terklon berukuran 698 pb sebagai daerah konservatif ENR tersebut mempunyai homologi tinggi dengan gen yang sama dari    O. sativa,  O. europaea, B. napus, T. aestivum dan  A. thaliana masing-masing dengan E-value 1e-96, 7e-77, 2e-64, 5e-41 dan 3e-36. Berdasarkan hasil tersebut telah dibuat primer spesifik untuk amplifikasi cDNA daerah ujung 5’- dan 3’- gen  ENR dari mesokarp kelapa 

scholarly journals Kloning parsial gen penyandi enoil-ACP reduktase dari mesokarp buah kelapa sawit (Elaeis guineensis Jacq.) Partial cloning of gene encoding enoyl-ACP reductase from mesocarp of oil palm (Elaeis guineensis Jacq.)

2016 ◽  
Vol 72 (1) ◽  
Author(s):  
Asmini BUDIANI ◽  
Djoko SANTOSO ◽  
Hajrial ASWIDINNOOR ◽  
Antonius SUWANTO
Keyword(s):  
Fatty Acid ◽  
Oil Palm ◽  
Fatty Acid Synthase ◽  
Elaeis Guineensis ◽  
Rt Pcr ◽  
Gene Encoding ◽  
Oil Accumulation ◽  
Enoyl Acp Reductase ◽  
Conserved Region ◽  
Elaeis Guineensis Jacq

Summary Enoyl-ACP reductase (ENR) is a component of fatty acid synthase (FAS) that is considered to play an important role in fatty acid elongation and oil accumulation of several plants. One of the proteins expressed coinciding with fruit development and oil accumulation in oil palm has been detected from the previous study and had homology with ENR. Therefore, as a part of genetic engineering program to improve oil content and quality in oil palm mesocarp, this research was aimed to clone cDNA conserved region of gene encoding enoyl-ACP reductase from oil palm mesocarp. Based on the amino acid sequence of the polypeptide that was homologous with ENR and combined with information of conserved region sequences of the same gene from other plant sources, primers were designed for amplifying conserved region of the ENR gene. Amplifi-cation was carried out by RT-PCR using total RNA as template, at several annealing temperatures and MgCl2 concentrations. Amplification product was cloned using pCR 2.1-TOPO, and the sequence was subjected into BlastN analysis. The results confirmed that the cloned cDNA fragment with 698 bp in size was the conserved region of the ENR gene.  This sequence was highly homologous with the same gene from other plants such as Oryza sativa, Olea europaea, Brassica napus, Triticum aestivum and Arabidopsis thaliana with E-value 1e-96, 7e-77, 2e-64, 5e-41 and 3e-36, respectively. Based on this result, primers have been made and used to amplify the 5’- and 3’ ends of the ENR -cDNA  of oil palm mesocarp. Ringkasan Enoil-ACP reduktase (ENR) merupakan salah satu komponen asam lemak sintase (FAS) yang berperan penting dalam pemanjangan asam lemak dan akumulasi minyak pada berbagai tanaman. Salah satu protein yang ter-ekspresi sejalan dengan tahapan perkembangan buah sawit dan akumulasi minyak pada penelitian sebelumnya diketahui mempunyai homologi dengan ENR. Oleh karena itu, sebagai salah satu bagian dari usaha rekayasa metabolisme minyak pada mesokarp buah sawit, penelitian ini bertujuan untuk mengklon cDNA daerah konservatif gen penyandi ENR dari mesokarp buah sawit. Berdasarkan  sekuen asam amino polipeptida yang mempunyai homologi dengan ENR dan dikombinasikan dengan hasil penjajaran daerah konservatif gen tersebut dari berbagai anaman lain, dirancang primer  untuk  amplifikasi daerah konservatif ENR. Amplifikasi dilakukan dengan RT-PCR menggunakan templat RNA total pada berbagai suhu   penempelan   dan   konsentrasi    MgCl2. Hasil amplifikasi dimurnikan dari gel dan diklon menggunakan vektor kloning pCR2.1-TOPO serta dianalisis nya menggunakan BlastN. Hasilnya mengkonfirmasi fragmen cDNA terklon berukuran 698 pb sebagai daerah konservatif ENR tersebut mempunyai homologi tinggi dengan gen yang sama dari    O. sativa,  O. europaea, B. napus, T. aestivum dan  A. thaliana masing-masing dengan E-value 1e-96, 7e-77, 2e-64, 5e-41 dan 3e-36. Berdasarkan hasil tersebut telah dibuat primer spesifik untuk amplifikasi cDNA daerah ujung 5’- dan 3’- gen  ENR dari mesokarp kelapa 

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