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1.
Unsaturated fatty acids having structural features which are different from those of the monoenoic acids normally synthesized by Escherichia coli can serve as growth factors for an auxotroph requiring unsaturated fatty acids. These analogues were incorporated into the phospholipids, as shown by gas-liquid and thin-layer chromatographic analysis of the phospholipid fatty acid composition. Some of these fatty acids were cisDelta(5)- and cis-Delta(9)-tetradecenoic, cis-Delta(11)-eicosenoic, cis,cis-Delta(11,14)-eicosadienoic, cis,cis,cis-Delta(11,14,17)-eicosatrienoic, trans-Delta(9)- and trans-Delta(11)-octadecenoic acids. Although partial degradation of some of these analogues to shorter even-chain homologues occurred, chain elongation of the exogenous fatty acids was not detected. Trans-olefinic acids were utilized without stereochemical or positional isomerization. These studies provide a basis for exploring the properties of the fatty acids and phospholipids required for the formation, structure, and function of membranes. 相似文献
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Saturated Fatty Acid Mutant of Saccharomyces cerevisiae with an Intact Fatty Acid Synthetase 总被引:5,自引:0,他引:5
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To determine directly the effects of streptomycin on translational fidelity in intact cells, we studied the synthesis of beta-galactosidase and of the coat protein of bacteriophage R17 in an Escherichia coli mutant in which the bactericidal effects of streptomycin are delayed. After the addition of streptomycin to exponentially growing mutant cells, protein synthesis continues at an undiminished rate for approximately an hour; however, as measured by enzyme assays, little functional protein is produced. Serological assays designed to detect beta-galactosidase and bacteriophage R17 coat protein show that substantial amounts of the protein synthesized can react with antisera prepared against active beta-galactosidase and phage R17, indicating the aberrance of the protein produced in the presence of the antibiotic. The polypeptides synthesized in the presence of streptomycin are degraded in the cell to a much greater extent than protein synthesized in the absence of the antibiotic. The proteolytic attack on this protein is not affected by inhibitors of serine proteases, suggesting that enzymes other than those involved in "normal turnover" of cellular protein are responsible. In this strain, certain of the multiple effects of streptomycin are separated in time and the production of abnormal protein (enzymatically inactive and susceptible to proteolytic attack) could be studied in the absence of the lethal effect of the drug. 相似文献
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Koichi Ogata Masahiro Osugi Tatsurokuro Tochikura 《Bioscience, biotechnology, and biochemistry》2013,77(10):1024-1029
During the investigation on the metabolism of azelaic acid by Micrococcus sp., it was found that the bacterium produced a large amount of keto acid (α-ketoglutaric acid) under the restricted condition for nitrogen source. The acid was identified as α-ketoglutaric acid by physico-chemical and biological methods. The mechanism of the production of α-ketoglutaric acid from azelaic acid was investigated. From the result, it was suggested that α-ketoglutaric acid production proceeded thrpugh the further oxidation of acetic acid produced from azelaic acid and that the production might be functioned by TCA cycle enzymes of the bacterium. Similarly, α-ketoglutaric acid was found to be produced remarkably from other various fatty acids. 相似文献
7.
Koichi Ogata Tatsurokuro Tochikura Masahiro Osugi Shojiro Iwahara 《Bioscience, biotechnology, and biochemistry》2013,77(2):176-180
The production of pimelic acid from azelaic acid by microorganisms was studied. About 100 strains of bacteria which were able to utilize azelaic acid as a sole carbon source were isolated from soil and other natural materials. Among these bacteria, several strains produced a large quantity of an organic acid (pimelic acid) from azelaic acid in their culture fluids during the cultivation. The acid was isolated from the culture fluid of strain A133 in crystalline form. The crystal was identified as pimelic acid by physicochemical and biological methods.From the results of investigations on the morphological and physiological characters, the bacterial strain A133 was assumed to be Micrococcus sp. 相似文献
8.
Zakim D 《The Journal of membrane biology》2000,176(2):101-109
There is continuing controversy about the mechanism for transfer of fatty acids (FA) between plasma and the interior of cells
and vice versa. One view is that this is a spontaneous process. The generally accepted view is that each step of the process is facilitated
by a specialized protein. Whether uptake is spontaneous or facilitated, the components of the uptake system, e.g., albumin,
water, FA, plasma membrane, and putative transport proteins of the plasma membrane, must behave according to the rules of
the physical chemistry of the system. We review these features to illustrate the constraints they impose on the design of
experiments to adduce the mechanism of uptake. Analysis of the literature in the context of the physical chemistry of the
uptake system indicates that arguments for a facilitated mechanism of uptake for FA are not supported by any data extant.
By contrast, comparison of the rates for individual steps of the pathway traversed by FA moving from albumin to the inside
of a cell (or vesicles of a model system) with rates of uptake of FA of tissues in the steady state shows that the rates of
the former are sufficient to account for the rate of the latter.
Received: 18 January 2000/Revised: 17 April 2000 相似文献
9.
In cyanobacteria fatty acids destined for lipid synthesis can be synthesized de novo, but also exogenous free fatty acids from the culture medium can be directly incorporated into lipids. Activation of exogenous fatty acids is likely required prior to their utilization. To identify the enzymatic activity responsible for activation we cloned candidate genes from Synechocystis sp. PCC 6803 and Synechococcus elongatus PCC 7942 and identified the encoded proteins as acyl-acyl carrier protein synthetases (Aas). The enzymes catalyze the ATP-dependent esterification of fatty acids to the thiol of acyl carrier protein. The two protein sequences are only distantly related to known prokaryotic Aas proteins but they display strong similarity to sequences that can be found in almost all organisms that perform oxygenic photosynthesis. To investigate the biological role of Aas activity in cyanobacteria, aas knockout mutants were generated in the background of Synechocystis sp. PCC 6803 and S. elongatus PCC 7942. The mutant strains showed two phenotypes characterized by the inability to utilize exogenous fatty acids and by the secretion of endogenous fatty acids into the culture medium. The analyses of extracellular and intracellular fatty acid profiles of aas mutant strains as well as labeling experiments indicated that the detected free fatty acids are released from membrane lipids. The data suggest a considerable turnover of lipid molecules and a role for Aas activity in recycling the released fatty acids. In this model, lipid degradation represents a third supply of fatty acids for lipid synthesis in cyanobacteria.Cyanobacteria present a diverse group of Gram-negative bacteria capable of oxygenic photosynthesis (Margulis, 1975). Their two photosystems, as well as other genetic and morphological similarities, identified them as putative predecessors of chloroplasts of eukaryotic plants (Wallace, 1982; Pakrasi, 1995). The structural similarities of cyanobacteria and chloroplasts are reflected in part by equivalence of biochemical pathways and their components. For instance, cyanobacterial fatty acid and glycerolipid compositions closely resemble those of the inner envelope and thylakoid membranes of chloroplasts (Roughan et al., 1980; Heinz and Roughan, 1983). In cyanobacteria, as well as in chloroplasts, fatty acids are synthesized by a type II fatty acid synthase (FAS) complex utilizing a freely dissociable acyl carrier protein (ACP; Froehlich et al., 1990). The products of FAS are released as acyl ACPs and may serve directly as substrates for acyltransferases, incorporating the fatty acids into membrane lipids (Frentzen et al., 1983). The substrate specificity of the acyltransferases establishes in cyanobacteria as well as in plastids the typical prokaryotic fatty acid pattern characterized by C16 fatty acids esterified to the sn-2 position. The correspondence of metabolic pathways between cyanobacteria and chloroplasts is reflected by the shared presence of closely related enzymes that catalyze key reactions. Besides the many similarities, however, there are also clear discrepancies that in part account for the fact that cyanobacteria are unicellular organisms, whereas chloroplasts are embedded in the metabolism of a eukaryotic cell. In terms of lipid metabolism, such differences become obvious if one considers the fact that the plastidial FAS also supplies the extraplastidic compartment with fatty acids (Browse et al., 1986). Fatty acid export from the chloroplast necessitates the release of synthesized acyl chains from ACP to allow transport across both envelope membranes. The release is achieved by the action of acyl-ACP thioesterases that hydrolyze the acyl-ACP thioester to liberate the fatty acid (Voelker et al., 1997). In cyanobacteria such export would obviously result in an unfavorable loss of fatty acids, and consequently homologous proteins to acyl-ACP thioesterases cannot be found here. Whereas cyanobacteria seem to be unable to release fatty acids enzymatically from their activated state, all cyanobacterial genomes available to date encode an activity most likely responsible for the activation of free fatty acids. The respective sequences are annotated as acyl-CoA synthetases. Conserved motifs in the amino acid sequence identify these proteins as members of the well-established superfamily of AMP-binding proteins. This protein family comprises several hundred amino acid sequences spreading across all organisms analyzed so far. The family members are annotated in the PROSITE database under entry number PS00455. Although these predicted fatty acid-activating enzymes of cyanobacteria are annotated as acyl-CoA synthetases due to their sequence similarity to proteins with such enzymatic activity, there is a much higher degree of similarity to certain AMP-binding proteins of plant origin with less-well-established function. These plant proteins are predicted to reside in chloroplasts and one member of this subgroup from Arabidopsis (Arabidopsis thaliana) designated as AAE15 was recently described as acyl-ACP synthetase. The conclusions were based on the comparison of enzymatic activity between plant extracts of wild-type and knockout mutant lines (Koo et al., 2005). Whereas the biological role of this activity remained largely elusive, it was shown that the capacity of plant extracts to elongate supplied medium fatty acids depended on AAE15 activity. Since the elongation of medium chain fatty acids in the plastid depends on the FAS requiring acyl ACPs, it was concluded that the fatty acids must have been activated by ACP. The elongated fatty acids ultimately appeared in membrane lipids. Together these findings suggested that AAE15 is an acyl-ACP synthetase.Besides encoding a protein homologous to AAE15 from Arabidopsis, cyanobacteria are also able to utilize exogenous fatty acids like it was shown for isolated chloroplasts. It is well established that feeding different cyanobacteria with free fatty acids results in the incorporation of these fatty acids into membrane lipids. For this process the activation of the fatty acids is believed to be essential. This causal relationship was clearly shown at least for other unicellular organisms like Escherichia coli and yeast (Saccharomyces cerevisiae) where the deletion of acyl-CoA synthetase activity resulted in the inability to utilize exogenous fatty acids (Overath et al., 1969; Knoll et al., 1995). It is not easy to assess how regularly cyanobacterial cells are exposed to exogenous free fatty acids in nature but at least for marine strains this is most likely a rather artificial situation. Therefore, it can be speculated that the capacity to activate free fatty acids might be of different relevance in the lipid metabolism of cyanobacteria in vivo.In this article, we investigated the fatty acid metabolism of cyanobacteria. We isolated candidate genes potentially encoding enzymes involved in fatty acid activation from the strains Synechocystis sp. PCC 6803 (hereafter Synechocystis) and Synechococcus elongatus PCC 7942 (hereafter Synechococcus) and performed heterologous expression in E. coli. The recombinant proteins were shown to possess acyl-ACP synthetase activity with broad substrate specificity. Knockout mutant strains deficient in acyl-ACP synthetase activity were characterized by secretion of endogenous free fatty acids into the culture medium. Combined with labeling experiments, the results suggest an essential role for acyl-ACP synthetase in fatty acid recycling in cyanobacteria. 相似文献
10.
Overproduction of a Functional Fatty Acid Biosynthetic Enzyme Blocks Fatty Acid Synthesis in Escherichia coli 总被引:3,自引:0,他引:3
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β-Ketoacyl-acyl carrier protein (ACP) synthetase II (KAS II) is one of three Escherichia coli isozymes that catalyze the elongation of growing fatty acid chains by condensation of acyl-ACP with malonyl-ACP. Overexpression of this enzyme has been found to be extremely toxic to E. coli, much more so than overproduction of either of the other KAS isozymes, KAS I or KAS III. The immediate effect of KAS II overproduction is the cessation of phospholipid synthesis, and this inhibition is specifically due to the blockage of fatty acid synthesis. To determine the cause of this inhibition, we examined the intracellular pools of ACP, coenzyme A (CoA), and their acyl thioesters. Although no significant changes were detected in the acyl-ACP pools, the CoA pools were dramatically altered by KAS II overproduction. Malonyl-CoA increased to about 40% of the total cellular CoA pool upon KAS II overproduction from a steady-state level of around 0.5% in the absence of KAS II overproduction. This finding indicated that the conversion of malonyl-CoA to fatty acids had been blocked and could be explained if either the conversion of malonyl-CoA to malonyl-ACP and/or the elongation reactions of fatty acid synthesis had been blocked. Overproduction of malonyl-CoA:ACP transacylase, the enzyme catalyzing the conversion of malonyl-CoA to malonyl-ACP, partially relieved the toxicity of KAS II overproduction, consistent with a model in which high levels of KAS II blocks access of the other KAS isozymes to malonyl-CoA:ACP transacylase. 相似文献
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海洋微藻脂肪酸去饱和酶 总被引:5,自引:0,他引:5
海洋微藻中富含多不饱和脂肪酸(polyunsaturated fatty acid,PUFA),在部分微藻中ω3 PUFA的量可达其总脂肪酸的30%~50%。而且微藻油具有鱼油所不可比拟的健康优势,也是唯一得到美国食品与药物管理局(FDA)认可的儿童DHA(二十二碳六烯酸)补充剂来源。由于用培养微藻来提取、纯化PUFA受到现有生产工艺的限制,使微藻油在国际食品(尤其是高质量食品)及保健品市场上供不应求。微藻脂肪酸去饱和酶(fatty aciddesaturase,FAD)是微藻PUFA合成的关键酶类,所以对微藻FAD的深入研究无疑将促进PUFA资源的合理开发和利用。 相似文献
12.
The rates of carboxylation, photophosphorylation and acetate incorporation have been compared in the intact and broken chloroplasts of Hydrilla verticillata Royle leaves in the presence and absence of certain inhibitors and metabolites. The intact chloroplasts showed low rates of photophosphorylation, high rates of carboxylation, and exhibited normal capacity for fatty acid biosynthesis. In broken chloroplasts a drastic decrease was observed in the rates of carboxylation and acetate incorporation. However, the rate of photophosphorylation was considerably increased. In the presence of light, inhibitors such as iodoacetamide, arsenite and sodium azide decreased the photophosphorylation rate. F-1,6-di-P and PGA stimulated CO2 fixation rate. In the absence of artificial light, inhibitors such as sodium arsenite, gluconate-6-phosphate, sodium azide and iodoacetamide decreased the rate of CO2 fixation. CoA, ATP, G-6-P, F-1,6-di-P Stimulated the synthesis of fatty acids. Exogenous supply of ADP. NADH, NADP and NADPH did not stimulate fatty acid biosynthesis probably because these compounds could not gain entry into the chloroplasts. Light was necessary for the in vitro fatty acid biosynthesis. 相似文献
13.
游离脂肪酸是机体的一种重要的能量物质,其代谢异常可以敏感的反应脂类代谢异常,是糖代谢紊乱、胰岛素抵抗发生发展的重要促进因素,也与高血压的发生发展明显相关.有研究发现在冠心病患者血清游离脂肪酸含量明显升高,游离脂肪酸参与冠状动脉发生过程中的氧化应激反应,参与破坏血管内皮细胞功能,促进冠心病的发生发展,并与冠心病心律失常等各种并发症的发生相关.作为心肌细胞代谢的能量底物,游离脂肪酸代谢异常在心衰患者的心肌细胞能量代谢障碍中起重要作用,心衰患者心肌细胞的能量代谢与正常成人的心肌细胞代谢出现明显差别,游离脂肪酸利用减少,而葡萄糖利用增加,现已经有应用阿昔莫司、乙克莫舍、哌克西林、曲美他嗪等药物通过调整游离脂肪酸代谢,从而改善心衰细胞心肌能量代谢的尝试.游离脂肪酸代谢异常的调控有望成为心血管疾病及其危险因素治疗及预防的新靶点. 相似文献
14.
Masayuki Mizuno Yukiji Shimojima Takashi Iguchi Isao Takeda Saburo Senoh 《Bioscience, biotechnology, and biochemistry》2013,77(5):506-510
As a part of extensive program on microbial utilization of hydrocarbons, lipid components of Candida petrophillum SD-14 grown on n-alkanes and glucose as carbon sources were studied. In any carbon source, cellular fatty acids of the yeast contained palmitic, palmitoleic, stearic, oleic and linoleic acids as major components.When n-tridecane was fed to the yeast, fatty acids with odd- and even-number of carbon atoms were produced in almost identical quantity. Another yeast, Torulopsis petrophillum SD-77, also gave a very similar fatty acid pattern by n-tridecane substrate. These phenomena indicate the existence of C2 addition and β-oxidation of the fatty acid formed in the yeasts.In the cases of n-tridecane, n-hexadecane and glucose as substrate, about a half of SD-14’s lipid was phospholipid, which consisted of phosphatidyl ethanolamine and phosphatidyl choline principally. Free alcohol and wax were not detected in any case. 相似文献
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Ronald Joseph 《Journal of bacteriology》1972,112(1):629-631
The fatty acid composition of Spirochaeta stenostrepta consists primarily of saturated, branch-chained fatty acids. Iso-C(15), anteiso-C(15), iso-C(17), and anteiso-C(17) represent 66% of the total fatty acids. 相似文献
17.
Wilkinson RE 《Plant physiology》1970,46(3):463-465
Sicklepod (Cassia obtusifolia L.) grew taller and heavier in a 12-hour photoperiod than in photoperiods longer or shorter. Total vegetative fatty acid content was maximal under 14-hour photoperiods as were unsaturated and branched chain fatty acids. Odd-numbered fatty acid content was highest under 16-hour photoperiods, and evennumbered fatty acids tended to be more concentrated under 14-hour photoperiods. Age of plant influenced total vegetative fatty acid content. 相似文献
18.
Silvia E. H. J. van Kempen Henk A. Schols Erik van der Linden Leonard M. C. Sagis 《Food biophysics》2014,9(2):114-124
In this article the effect of variations in the fatty acid chain of oligofructose fatty acid esters (OFAE) on foamability and foam stability is described. First, oligofructose (OF) mono-esters containing saturated fatty acid chains ranging between C4 and C18 were studied. Additionally, a mono-ester containing a C16 mono-unsaturated fatty acid chain and a C12 di-ester were studied. Finally, to investigate the influence of the size of the hydrophilic group, commercially available sucrose esters were studied. The surface tension and surface rheological properties of air/water interfaces stabilized by the esters were determined, as well as the foaming properties of the esters, at a bulk concentration of 0.2 % (w/v). OF mono-esters with intermediate fatty acid chain lengths (C10-C16) were able to migrate quickly to the interface producing foams with small bubbles (0.4 mm), a relatively narrow bubble size distribution, and a high stability. For oligofructose mono-esters containing fatty acids C4 and C8, the bulk concentration of 0.2 % (w/v) was below the CMC, resulting in insufficient surface coverage, and low foamability and foam stability. The OF C18 mono-ester and the OF C12 di-ester were slow to migrate to the interface resulting in low foamability. Despite similar surface tension values, the foam half-life time of OFAE was higher than of the corresponding sucrose esters. OFAE gave higher surface dilatational moduli compared to sucrose esters. Based on the frequency dependence of the modulus and analysis of Lissajous plots, we propose that OFAE may be forming a soft glass at the interface. 相似文献
19.
Control of Fatty Acid Metabolism I. Induction of the Enzymes of Fatty Acid Oxidation in Escherichia coli 总被引:11,自引:29,他引:11
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Escherichia coli grows on long-chain fatty acids after a distinct lag phase. Cells, preadapted to palmitate, grow immediately on fatty acids, indicating that fatty acid oxidation in this bacterium is an inducible system. This hypothesis is supported by the fact that cells grown on palmitate oxidize fatty acids at rates 7 times faster than cells grown on amino acids and 60 times faster than cells grown on a combined medium of glucose and amino acids. The inhibitory effect of glucose may be explained in terms of catabolite repression. The activities of the five key enzymes of beta-oxidation [palmityl-coenzyme A (CoA) synthetase, acyl-CoA dehydrogenase, enoyl-CoA hydrase, beta-hydroxyacyl-CoA dehydrogenase, and thiolase] all vary coordinately over a wide range of activity, indicating that they are all under unit control. The ability of a fatty acid to induce the enzymes of beta-oxidation and support-growth is a function of its chain length. Fatty acids of carbon chain lengths of C(14) and longer induce the enzymes of fatty acid oxidation and readily support growth, whereas decanoate and laurate do not induce the enzymes of fatty acid oxidation and only support limited growth of palmitate-induced cells. Two mutants, D-1 and D-3, which grow on decanoate and laurate were isolated and were found to contain constitutive levels of the beta-oxidation enzymes. Short-chain fatty acids (相似文献
20.
Control of Fatty Acid Synthesis in Bacteria 总被引:12,自引:5,他引:7
Leonard Mindich 《Journal of bacteriology》1972,110(1):96-102
When glycerol-requiring auxotrophs of Bacillus subtilis are deprived of glycerol, the synthesis of fatty acids continues at an apparent rate of 20 to 50% that of supplemented cultures. The newly synthesized fatty acids are not incorporated into phospholipid and accumulate as free fatty acids. These molecules undergo a much more rapid turnover than phospholipid fatty acids, and the rate of turnover is sufficient to indicate that the rate of fatty acid synthesis in glycerol-deprived cultures is similar to that in supplemented ones. The average chain length of the free fatty acids is greater than that of the phospholipid fatty acids. Cells deprived of required amino acids also show a diminution in the apparent rate of fatty acid synthesis; however, in this case, the fatty acids accumulate in phospholipid, and no increase of the free fatty acid fraction is observed. It is argued on the basis of these findings that the control of lipid synthesis does not operate at the level of transacylation but must act on one or more of the reactions of the fatty acid synthetase. 相似文献