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1.
Fatty acid biosynthesis in the leaves of barley, wheat and pea.   总被引:5,自引:1,他引:4       下载免费PDF全文
1. The incorporation of radioactivity from [1-14C]acetate into the leaf lipids of barley, pea and wheat has been studied in pulse-labelling experiments. 2. There was little increase in the total labelling of lipids after the leaves were transferred to non-radioactive medium. However, there was an increase in the relative labelling of unsaturated fatty acids. In addition, there was an increase in the relative labelling of diacylgalactosylglycerol. 3. The principal radioactively labelled acyl lipids were diacylgalactosylglycerol and phosphatidylcholine. Phosphatidylcholine showed a decreasing proportion of [14C]oleate and an increasing amount of [14C]linoleate with time. Diacylgalactosylglycerol also had decreasing amounts of [14C]oleate but, in addition, had an increasing proportion of [14C]linolenate with time. 4. The absence of significant amounts of [14C]linolenate in phosphatidylcholine appeared to exclude a role for this phospholipid in linoleate desaturation. 5. The specific radioactivities of oleate and linoleate in phosphatidylcholine, diacylgalactosylglycerol and diacylgalabiosylglycerol were very similar in any single experiment. It was concluded that these fatty acids can rapidly exchange between the three intact lipids.  相似文献   

2.
1. The patterns of incorporation of (14)C into glycerolipid fatty acids of developing maize leaf lamina from supplied [1-(14)C]acetate and from (14)CO(2) during steady-state photosynthesis were similar. Oleate of phosphatidylcholine and palmitate of phosphatidylglycerol attained linear rates of labelling more rapidly than did other fatty acids, particularly the linoleate and linolenate of monogalactosyl diacylglycerol. 2. After the transfer of lamina from labelled to unlabelled acetate, there was a decrease in labelled oleate and linoleate of phosphatidylcholine and a concomitant increase in the amount of radioactivity in the linoleate and linolenate of monogalactosyl diacylglycerol. 3. The rapidly labelled phospholipids, phosphatidylcholine and phosphatidylglycerol, were shown by differential and sucrose-density-gradient centrifugation to be associated with different organelles, the former being mainly in a low-density membrane fraction, probably microsomal, and the latter mainly in chloroplasts. 4. During a 48h period after supplying spinach leaves with [(14)C]acetate, radioactivity was lost from the oleate of phosphatidylcholine present in fractions sedimented at 12000g and 105000g, and accumulated in the linolenate of monogalactosyl diacylglycerol of the chloroplast. 5. It is proposed that the phosphatidylcholine of some non-plastid membranes is intimately involved in the process of oleate desaturation and that this lipid serves as a donor of unsaturated C(18) fatty acids to other lipids, principally monogalactosyl diacylglycerol, of the chloroplasts.  相似文献   

3.
[1-14C]Oleic and [1-14C]linoleic acids were rapidly desaturated when incubated with maize leaves from 8-day-old plants and the labeled fatty acids, and their desaturation products, were rapidly incorporated into glycerolipids. Oleic acid was desaturated to linoleate at the rate of 0.7 nmol/100 mg tissue/h and further desaturated to linolenate at about one-third this rate. The rates of linolenate formation were similar when either oleic acid or linoleic acid was the substrate although there was a 2-h lag period when oleic acid was substrate. When radioactive oleic, linoleic, and linolenic acids were substrates, phosphatidylcholine was the most extensively labeled glycerolipid followed by monogalactosyldiacylglycerol. The relative rates of incorporation of label into individual glycerolipids are consistent with a movement of labeled fatty acids from phosphatidylcholine to monogalactosyldiacylglycerol and then to diagalactosyldiacylglycerol. The rates of labeling of phosphatidylcholine oleate and of phosphatidylcholine linoleate are consistent with a precursor-product relationship in that there was a delayed accumulation of phosphatidylcholine linoleate relative to that of phosphatidylcholine oleate and phosphatidylcholine linoleate continued to accumulate while phosphatidylcholine oleate declined. Linoleate formed from oleate was widely distributed in glycerolipids but neither phosphatidylcholine linolenate nor linolenate-containing diacylglycerol was detected at short and intermediate incubation times when either oleic or linoleic acid was substrate. The kinetics of incorporation of linoleate and linolenate into monogalactosyldiacylglycerol suggest a transfer of linoleate from phosphatidylcholine. The initial rate of accumulation of labeled linolenate in monogalactosyldiacylglycerol was very similar to the rate of desaturation of linoleate and it is suggested that desaturation of linoleate occurs while associated with monogalactosyl-diacylglycerol.  相似文献   

4.
The patterns of incorporation of [l-14C]-acetate into the glycerolipid fatty acids of leaves of olive plants ( Olea europea L. cv. Chétoui) suggested a specific pathway for a-linolenic acid biosynthesis. The results confirmed the involvement of phosphati-dylcholine in galactolipid metabolism, and seemed to exclude the role of that mole-cule as a substrate for desaturation of oleate to linoleate. The two oleate desaturation steps seemed to occur rapidly on the diacylgalactosylglycerol molecule for biosynthesis of galactolipid linolenate. In addition, the results indicated a slow sequen-tial desaturation of oleate to linolenate via linoleate in the phospholipid molecules (phosphatidylcholine, phosphatidylethanolamine and phosphatidylglycerol).  相似文献   

5.
Exogenous [1-14C]oleic acid and [1-14C]linoleic acid were taken up and esterified to complex lipids by greening cucumber (Cucumis sativus L.) cotyledons. Both 14C-labeled fatty acids were initially esterified to phosphatidylcholine prior to eventual accumulation in triacylglycerols and galactolipids. Kinetic data suggest that esterification occurs prior to desaturation and that phosphatidylcholine is the initial site of both [14C]-oleate and [1-14C]linoleate esterification and of [1-14C]oleate desaturation to [1-14C]linoleate. [1-14C]Linoleic acid was esterified more rapidly than [14C]oleic acid and its desaturation product, [1-14C]α-linolenate, occurred mainly on monogalactosyl diacylglycerol, although some was also observed on the other major acyl lipids, including phosphatidylcholine.  相似文献   

6.
The effects of water stress on [1-14C]-oleic and [1-14C]-linoleic acid desaturations were studied in leaves of two varieties of cotton ( Gossypium hirsutum L.), one drought-sensitive (Reba) and the other more resistant (Mocosinho). After 24 h incorporation, [1-14C]-oleate led to the appearance of linoleate in phospholipids and, additionally, of linolenate in galactolipids. [1-14C]-Linoleate was desaturated to linolenate only in galactolipid fractions. Water stress markedly inhibited the incorporation of the precursors into the leaf lipids. The two desaturation steps were affected, particularly the transformation of linoleate to linolenate in monogalactosyldiacylglycerol in the drought-sensitive variety of cotton. The metabolic implications of the inhibition of the biosynthesis of C18-polyunsaturated fatty acids are discussed.  相似文献   

7.
Hawke JC  Stumpf PK 《Plant physiology》1980,65(6):1027-1030
Oleate and linoleate desaturation in leaves of maize seedlings was largely independent of previous light treatment of the seedlings; there was no evidence of light-induced desaturase activities. These results are in sharp contrast to those observed with developing cucumber cotyledons in which pronounced increase in desaturation occurs after exposure of tissue to light. The rates of desaturation of oleate were about four times those of linoleate in both etiolated and 16-hour greened maize leaves. In both etiolated and greened tissues, about two-thirds of the label from oleate was esterified after 4 hours, half of which was in phosphatidylcholine. Phosphatidylcholine and diglyceride contained large proportions of [14C]linoleate formed from [14C]oleate but not [14C]linolenate. In monogalactolipid, about two-thirds of the labeled fatty acids were linolenate. In vivo desaturase activity was present in tissue of widely different levels of differentiation and chlorophyll content obtained from light-grown maize seedlings.  相似文献   

8.
9.
Phosphorylase kinase phosphorylation of skeletal-muscle troponin T.   总被引:1,自引:1,他引:0       下载免费PDF全文
When [14C]diacylgalactosylglycerol was added to isolated pea or lettuce chloroplasts linolenate synthesis was seen. The desaturation of [14C]linoleate in diacylgalactosylglycerol to [14C]linolenate was stimulated by the addition of a soluble protein fraction containing lipid-exchange activity. Other [14C]acyl lipids were ineffective, except that [14C]phosphatidylcholine in the presence of UDP-galactose and sn-glycerol 3-phosphate could also supply [14C]linoleate for desaturation. These results are consistent with a role of diacylgalactosylglycerol in linolenate synthesis, as indirectly suggested by labelling experiments.  相似文献   

10.
Microsomal membrane preparations from the immature cotyledons of safflower (Carthamus tinctorius) catalysed the interconversion of the neutral lipids, mono-, di-, and triacylglycerol. Membranes were incubated with neutral lipid substrates, 14C-labelled either in the acyl or glycerol moiety, and the incorporation of radioactivity into other complex lipids determined. It was clear that diacylglycerol gave rise to triacylglycerol and monoacylglycerol as well as phosphatidylcholine. Radioactivity from added [14C] triacylglycerol was to a small extent transferred to diacylglycerol whereas added [14C] monoacylglycerol was rapidly converted to diacylglycerols and triacylglycerols. The formation of triacylglycerol from diacylglycerol occurred in the absence of acyl-CoA and hence did not involve diacylglycerol acyltransferase (DAGAT) activity. Monoacylglycerol was not esterified by direct acylation from acyl-CoA. We propose that these reactions were catalyzed by a diacylglycerol: diacylglycerol transacylase which yielded triacylglycerol and monoacylglycerol, the reaction being freely reversible. The specific activity of the transacylase was some 25% of the diacylglycerol acyltransferase activity and, hence, during the net accumulation of oil, substantial newly formed triacylglycerol equilibrated with the diacylglycerol pool. In its turn the diacylglycerol rapidly interconverted with phosphatidylcholine, the major complex lipid substrate for Δ12 desaturation. Hence, the oleate from triacylglycerols entering phosphatidylcholine via this route could be further desaturated to linoleate. A model is presented which reconciles these observations with our current understanding of fatty acid desaturation in phosphatidylcholine and oil assembly in oleaceous seeds. Received: 8 November 1996 / Accepted: 5 February 1997  相似文献   

11.
Developing cotyledons of safflower (Carthamus tinctorius L.) and sunflower (Helianthus annuus L.) readily utilised exogenously supplied 14C-labelled fatty-acid substrates for the synthesis of triacylglycerols. The other major radioactive lipids were phosphatidylcholine and diacylglycerol. In safflower cotyledons, [14C]oleate was rapidly transferred to position 2 of sn-phosphatidylcholine and concomitant with this was the appearance of radioactive linoleate. The linoleate was further utilised in the synthesis of diacyl- and triacyl-glycerol via the reactions of the so-called Kennedy pathway. Supplying [14C]linoleate, however, resulted in a more rapid labelling of the diacylglycerols than from [14C]oleate. In contrast, sunflower cotyledons readily utilised both labelled acyl substrates for rapid diacylglycerol formation as well as incorporation into position 2 of sn-phosphatidylcholine. In both species, however, [14C]palmitate largely entered sn-phosphatidylcholine at position 1 during triacylglycerol synthesis. The results support our previous in-vitro observations with isolated microsomal membrane preparations that (i) the entry of oleate into position 2 of sn-phosphatidylcholine, via acyl exchange, for desaturation to linoleate is of major importance in regulating the level of polyunsaturated fatty acids available for triacylglycerol formation and (ii) Palmitate is largely excluded from position 2 of sn-phosphatidylcholine and enters this phospholipid at position 1 probably via the equilibration with diacylglycerol. Specie differences appear to exist between safflower and sunflower in relation to the relative importance of acyl exchange and the interconversion of diacylglycerol with phosphatidylcholine as mechanisms for the entry of oleate into the phospholipid for desaturation.Abbreviations FW fresh weight - TLC thin-layer chromatography  相似文献   

12.
ABSTRACT. Major fatty acid components of Acanthamoeba castellanii lipids extracted after growth at 30°C include myristate, palmitate, stearate and the polyunsaturates linoleate, eicosadienoate, eicosatrienoate and arachidonate, with oleate as the sole major monounsaturated fatty acid. By comparison, growth at 15°C gave increased linoleate, eicosatrienoate and arachidonate, but decreased oleate and palmitate. When the growth temperature was shifted downwards from 30°C to 15°C, increased lipid unsaturation occurred over a period of 24 h; thus decreases of oleate and eicosadienoate were accompanied by increases in linoleate, eicosatrienoate, arachidonate and eicosapentaenoate. An upwards shift from 15°C to 30°C gave negligible alterations in fatty acid composition over a similar period. At 15°C organisms rapidly use [1-14C] acetate for de novo fatty acid synthesis; stearate is converted via oleate to further desaturation and chain elongation products. Similar short term experiments at 30°C indicate only de novo synthesis and Δ9-desaturation; synthesis of polyunsaturates was a much slower process. Rapid incorporation of [1-14C] oleate at 30°C was not accompanied by metabolic conversion over two hours, whereas at 15°C n-6 desaturation to linoleate was observed. Temperature shift of organisms from 15°C to 30°C in the presence of [1-14C] acetate revealed that over half of the fatty acids in newly-synthesised lipids were saturated, but the proportions of unsaturated fatty acids increased with time until the total polyenoate components reached 17% after 22 h. A shift of temperature in the reverse direction gave a corresponding figure of 60% for polyunsaturated fatty acids. These results emphasize the importance of n-6 desaturation in the low temperature adaptation of Acanthamoeba castellanii .  相似文献   

13.
Etiolated Cucumis sativus L. cotyledons preferentially catabolized exogenous [1-14C]oleic acid and [1-14C]linoleic acid with relatively little incorporation into complex lipids or desaturation of the 14C-labeled fatty acids. Following a 16-hour exposure to light, the greening cotyledons efficiently desaturated the exogenous 14C-labeled fatty acids. A small amount of oleate desaturation to linoleate was observed in etiolated tissue, but hardly any linoleate desaturation to α-linolenate was detected. Both oleate and linoleate desaturation showed diurnal variations with maxima at the end of light periods and minima at the end of dark periods. Illumination of etiolated tissue by flashing light, as opposed to continuous light, failed to stimulate either chlorophyll or α-linolenic acid biosynthesis, and both processes could be halted or reversed by 10 micrograms per milliliter cycloheximide. Production of polyunsaturated fatty acids from [1-14C]acetate, [1-14C]oleic acid, and [1-14C]linoleic acid, by greening cucumber cotyledons, was markedly affected by tissue integrity with finely chopped cotyledons having very little capacity for their synthesis and intact seedlings showing the highest rates.  相似文献   

14.
Microsomes isolated from the developing cotyledons of the seeds of the safflower varieties, very-high-linoleate, Gila and high-oleate, were capable of exchanging the acyl groups in acyl-CoA with the fatty acids in position 2 of phosphatidylcholine. The specificity of the 'acyl-exchange' towards the acyl moiety in acyl-CoA was selective in the order: oleate greater than linoleate greater than linolenate. Stearoyl-CoA was completely selected against when presented in a mixed substrate with unsaturated 18-carbon acyl-CoAs. Microsomes, of the very-high-linoleate safflower variety, rapidly desaturated in situ-labelled [14C]oleoylphosphatidylcholine in the presence of NADH. Little oleate desaturation, however, was observed in the microsomes of the high-oleate variety. Microsomes of the Gila and high-oleate varieties of safflower rapidly synthesised phosphatidic acid by the acylation of glycerol 3-phosphate with acyl-CoA. The phosphatidic acid was metabolised to diacylglycerol, which was further acylated to triacylglycerol. A strong selectivity for linoleoyl-CoA was found for the acylation of glycerol 3-phosphate in both the Gila and high-oleate microsomes. On the basis of these results, we propose that the pattern of 18-carbon unsaturated fatty acids in the triacylglycerols of all 'oil'-producing seeds is a direct reflection of the fatty acids in the acyl-CoA pool. This, in turn, is governed by: A, the rate and specificity of the acyl exchange between acyl-CoA and phosphatidylcholine; B, the rate of oleate (and linoleate) desaturation in phosphatidylcholine; and C, the rate and specificity of the glycerophosphate acyltransferase.  相似文献   

15.
1. Surgically prepared lactating goats were used to obtain quantitative information on the biohydrogenation and absorption of dietary fat, and on the mammary uptake and transfer into milk fat of the complex mixture of cis- and trans-isomers of octadecenoate that arise during ruminal biohydrogenation. 2. About 90% of dietary linolenate, linoleate and oleate was hydrogenated in the rumen, and the availability to the animals of the essential fatty acid, linoleate, represented only 0.5-1.5% of the total dietary energy. 3. The intra-ruminal administration of (14)C-labelled linolenate and linoleate showed that these acids were not absorbed from the rumen, in agreement with previous work. 4. No selectivity was observed in the metabolism of the geometrical and positional isomers of octadecenoate: their rates of absorption from the small intestine, transfer into lymph, uptake by the mammary gland and appearance in milk fat were similar. 5. The desaturase activity of intestinal epithelium was demonstrated by the appearance in lymph of [1-(14)C]oleate after the addition of [1-(14)C]stearate to the small intestine.  相似文献   

16.
Human skin fibroblasts incorporate and actively desaturate long-chain fatty acids. Growth of these cells in lipid-free medium can be used to enhance delta 9 and delta 6 desaturation of [14C]stearate and [14C]linoleate, respectively. Medium supplementation with cis fatty acids inhibits delta 9 desaturation; effectiveness as inhibitors is linoleate (9c,12c-18:2) greater than oleate (9c-18:1) greater than vaccenate (11c-18:1). Linoelaidate (9t,12t-18:2), trans-vaccenate (11t-18:1) and saturated fatty acids are without effect; elaidate (9t-18:1) appears stimulatory. By contrast, the trans fatty acids elaidate and linoelaidate are potent inhibitors of delta 6 desaturation; inhibition by trans-vaccenate is 50% of that of elaidate. Desaturation of [14C]linoleate is only slightly inhibited by oleate, cis-vaccenate, or (6c,9c,12c)-linolenate. The relative effectiveness of isomeric cis- and trans-octadecenoic acids as inhibitors of delta 9 and delta 6 desaturation in intact human cells is different from that found in microsomal studies. The cell culture system can thus be important in evaluating physiological effects of isomeric fatty acids on cellular metabolic processes.  相似文献   

17.
Evidence was obtained that Penicillium chrysogenum can produce linolenate by two biosynthetic pathways, i.e., by elongation of a shorter trienoic acid as well as direct desaturation of 18-C acids. In oxygen deficient cultures, exogenous hexadecatrienoate stimulated [1-14C]acetate incorporation into labeled octadecatrienoate and [U-14C]hexadecatrienoate with nonlabeled acetate yielded linolenate that had relatively little label in the 1-C position. With [1-14C]acetate as the only added substrate, oxygen deficiency inhibited incorporation of label into monoenoic and dienoic acids but not into trienoic acids. Incorporation of the [U-14C]linoleate into linolenate also was inhibited.In aerated cultures, 1-14C-label from laurate, palmitate, stearate, oleate, linoleate, and hexadecatrienoate was readily incorporated into linolenate. Decarboxylation and oxidation studies indicated that the longer acids were incorporated largely intact. [U-14C]Linoleate was incorporated into linolenate in which the fraction of label in 1-C was similar to that of the substrate. These data suggest that this mold has broader synthetic capabilities than do some chloroplast systems for the biosynthesis of linolenate.  相似文献   

18.
The total seed lipids of four flax (Linum usitatissimum) genotypes, differing markedly in their acyl composition, were extracted and fractionated using column, preparative, and thin-layer chromatography. In the total lipid extract of seeds, the lower linolenate content of the cultivar Glenelg (39.1% compared to that of cv. Croxton (50.5%) was associated with a higher oleate content. Further reductions in linolenate content in the induced mutants of cv. Glenelg, M1722 (17.2%) and "Zero" (1.9%) were accompanied by equivalent increases in linoleate but only minor increases in oleate. Similar changes were observed in the major triacylglycerol fraction of the simple lipids (fatty acid esters of glycerol and sterols), but there was considerable heterogeneity for acyl composition in the minor simple lipid components, including both diacylglycerols and sterol esters, and the complex lipids (glycolipids and phospholipids). The induced mutations substantially reduced linolenate content of all lipid fractions but in no case was it eliminated. Maturation of "Zero" seed at 15/10 degrees C (compared to 24/19 degrees C) increased linoleate and decreased stearate and oleate contents in all lipid fractions. In contrast to seed lipids, the acyl composition of the leaf lipids of the mutant genotypes was the same as those of their parent.  相似文献   

19.
The developing seeds of Borago officinalis (common borage) accumulate a triacylglycerol oil that is relatively rich in the uncommon fatty acid gamma-linolenate (octadec-6,9,12-trienoic acid). Incubation of developing, whole, cotyledons with [14C]oleate and [14C]linoleate showed that the gamma-linolenate was synthesized by the sequential desaturation of oleate----linoleate----gamma-linolenate. Microsomal membrane preparations from the developing cotyledons contained an active delta 6-desaturase enzyme that catalysed the conversion of linoleate into gamma-linolenate. Experiments were designed to manipulate the [14C]linoleate content of the microsomal phosphatidylcholine. The [14C]linoleoyl phosphatidylcholine labelled in situ was converted into gamma-linolenoyl phosphatidylcholine in the presence of NADH. The substrate for the delta 6-desaturase in borage was, therefore, the linoleate in the complex microsomal lipid phosphatidylcholine, rather than, as in animals, the acyl-CoA. This was further confirmed in experiments that compared the specific radioactivity of the gamma-linolenate, in acyl-CoA and phosphatidylcholine, that was synthesized when [14C]linoleoyl-CoA was incubated with microsomal membranes, NADH and non-radioactive gamma-linolenoyl-CoA. The delta 6-desaturase was positionally specific and only utilized the linoleate in position 2 of sn-phosphatidylcholine. Analysis of the positional distribution of fatty acids in the endogenous microsomal sn-phosphatidylcholine showed that, whereas position 1 contained substantial linoleate, only small amounts of gamma-linolenate were present. The results shed further light on the synthesis of C18 polyunsaturated fatty acids in plants and in particular its relationship to the regulation of the acyl quality of the triacylglycerols in oilseeds.  相似文献   

20.
The effects of the addition of hypolipidemic drugs and 1-acylglycerolipids on the metabolism of oleate in plants have been studied in vivo and in vitro. Using aged potato slices with [14C]oleate as a precursor, it was found that these drugs markedly inhibited both the incorporation into complex lipids and the desaturation of oleate to linoleate. Moreover, in vitro experiments, carried out with microsomes prepared from developing safflower seeds and [14C]oleate or [14C]oleoyl-CoA as precursors, confirmed the inhibitory effect of the drugs on oleate desaturation, and showed that while WY14643 mainly affected oleoyl thiokinase activity, DH990 exerted its strongest effect on the formation of PL, indicating that the mode of action of these two drugs in safflower microsomes is essentially different. Addition of LPC or LPE stimulated the incorporation of radiolabeled precursor into PC and PE, respectively, as well as the desaturation of oleate to linoleate when [14C]oleoyl-CoA was the precursor. The evidence obtained suggests that oleoyl-PE, as well as oleoyl-PC, should be considered as a possible substrate for oleate desaturation in plants.  相似文献   

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