Relationship among Cellular Fatty Acid Composition,Amino Acid Uptake and Neomycin Formation in a Mutant Strain of Streptomyces fradiae

Neomycin-producing strains of Streptomyces fradiae, whose cellular fatty acid spectra are of iso 16: 0-type or normal 16: 0-type, had about two to six times larger amino acid and hexosamine pools than a neomycin-nonproducing strain which has the anteiso 15: 0-type cellular fatty acid spectrum. About 50 to 80 percent of the amount of extractable free amino acids were L-glutamic acid in either type of cells. The difference of pool size in these strains seems to be explained by the difference in ability for amino acid uptake. That is, the ability for L-glutamate uptake of anteiso 15:0-type cells was markedly reduced and accumulated glutamate was easily washed out by buffer. Glucose, magnesium ions and L-glutamate were essential for the formation of neomycin by washed cells and, therefore, even the mutant ST–5B of anteiso 15: 0-type could accumulate a large amount of glutamate and produce neomycin as far as it was grown in a medium containing a high concentration of glutamate. These results indicate that a large pool of glutamate is essential for the formation of neomycin and the fatty acid spectrum is a factor governing the capacity to accumulate L-glutamic acid.     

费氏链霉菌中子辐射诱变菌株对新霉素的生物转化

对新霉素产生菌费氏链霉菌进行中子辐射诱变和突变株筛选, 获得不产新霉素的突变株。以突变株为转化菌种, 以新霉素为底物, 对转化发酵液进行高效液相色谱分析, 研究了不同转化条件对新霉素转化的影响。结果表明, 底物浓度、底物添加时间、底物添加方式、接种量、培养基装量、转化时间、碳源、氮源、pH、温度对新霉素转化具有不同程度的影响。以转化条件优化参数进行转化大量培养, 转化液经4步离子交换层析进行分离纯化, 薄层层析检测纯化样品为单一斑点。采用薄层生物自显影对获得的4个转化产物分离样品做生物活性检测, 发现4个样品对金黄色葡萄球菌和姜青枯假单孢杆菌都具有抑制活性, 1个样品对大白菜软腐样品具有明显的抑制活性。     

The Manipulation of the Fatty Acid Composition of Glycerolipids in Cyanobacteria Using Exogenous Fatty Acids

The fatty acid composition of the major diacylglycerolipidsof Anacystis nidulans R2 cells was modified by the careful additionof exogenous fatty acids to the growth medium of the cyanobacteria.In this way it is possible to incorporate significant levelsof different fatty acids into the membranes of the cells andchange their physical properties without changing other environmentalconditions. The exogenous fatty acid reduces or inhibits thebiosynthesis of native fatty acids and substitutes for themin all the major diacylglycerolipids. Although modifying thefatty acid composition in this way is lethal above certain concentrations,the data demonstrate a remarkable ability of the cells to growand develop over a wide range of fatty acid compositions. Similardata show that the fatty acid composition of Synechocystis sp.6803 cells also undergo significant changes in fatty acid compositionon the addition of exogenous fatty acid to the growth mediumand can tolerate similar changes in the degree of unsaturationof the fatty acids of the diacylglycerolipids of the cell membranes. (Received September 4, 1989; Accepted March 13, 1990)     

Effects of Minerals on Neomycin Production by Streptomyces fradiae

A study was made on the mineral requirements of Streptomyces fradiae strain 3535 for neomycin production. It was observed that optimal levels of the elements Ca, Fe, and Zn per milliliter of a synthetic medium for neomycin production were 10.8, 1.0, and 0.115 μg, respectively. K₂HPO₄ was required at a concentration of 0.1% for maximal yield of neomycin, whereas NaCl and the metals Mn and Cu were without any effect. High doses of Zn (0.23 μg/ml or above) caused destruction of neomycin after the fifth day of fermentation.     

Effect of Temperature on Fatty Acid Composition of a White Thermus Strain

A white Thermus sp. strain, NCIMB 11245, showed high levels of anteiso C_17:0, anteiso C_17:1, normal C_16:1, and iso C_16:0 with low levels of iso C_15:0 + iso C_17:0 in comparison to yellow-pigmented strains. The fatty acid composition may be associated with precursor metabolism or the absence of carotene pigmentation.     

Effect of Dietary Mono- and Polyunsaturated Fatty Acids on the Fatty Acid Composition of Pigs' Adipose Tissues

In two experiments with growing-finishing pigs six different dietary fats were added to a conventional diet (control - C) to study the effects of dietary monounsaturated (MUFA) and polyunsaturated fatty acids (PUFA) on the fatty acid composition of backfat and kidney fat at similar amounts of double bonds in feed (Exp. 1:7% pork fat - PF, 4.95% olive oil - OO, 3.17% soybean oil - SO) or a constant amount of 5% of processed fats (Exp. 2: partially hydrogenated fat - SAT, fractionated pork fats: olein - OLE, stearin - STE). Compared with the control, PUFA were only slightly increased in backfat of pigs fed PF, OLE, STE or OO, although dietary PUFA intake was up to 70% higher. With SO PUFA were significantly increased in adipose tissues, predominantly at the expense of MUFA. Consequently, a non-linear relationship was found between PUFA intake and proportion in backfat. MUFA were incorporated at the expense of SFA, therefore, adipose tissues of OO fed animals were lowest in SFA. Despite comparable amounts of double bonds in feed (Exp. 1), the degree of unsaturation measured as fat score (sum of double bonds) was in the order SO > OO > PF > C. In contrast, the proportion of SFA was C > PF = SO > OO. Regarding the decisive role of SFA for fat consistency it may be concluded that MUFA should also be considered in feeding recommendations for pigs. Furthermore, in case of a high dietary supply of MUFA, a simple index of double bonds might not be sufficiently conclusive to judge pig fat quality.     

Effect of Aromatic Compounds on Cellular Fatty Acid Composition of Rhodococcus opacus

In cells of Rhodococcus opacus GM-14, GM-29, and 1CP, the contents of branched (10-methyl) fatty acids increased from 3% to 15 to 34% of the total fatty acids when the cells were grown on benzene, phenol, 4-chlorophenol, chlorobenzene, or toluene as the sole source of carbon and energy, in comparison with cells grown on fructose. In addition, the content of trans-hexadecenoic acid increased from 5% to 8 to 18% with phenol or chlorophenol as the carbon source. The 10-methyl branched fatty acid content of R. opacus GM-14 cells increased in a dose-related manner following exposure to phenol or toluene when toluene was not utilized as the growth substrate. The results suggest that 10-methyl branched fatty acids may participate in the adaptation of R. opacus to lipophilic aromatic compounds.     

Utilization of Carbon- and Nitrogen-containing Compounds for Neomycin Production by Streptomyces fradiae

A number of carbon and nitrogen compounds were tested for their effect on growth of Streptomyces fradiae 3535 and neomycin production. Dextrin, starch, and maltose were excellent carbon sources for neomycin production, and sodium nitrate, aspartic acid, and glutamic acid were adequate nitrogen sources. Studies on biochemical changes during fermentation in four typical media indicated that there was no direct relation between the growth of the organism and neomycin formation. The pH of the medium might be an important factor for neomycin synthesis. The quantitative formation of neomycin components depended on the variation of carbon and nitrogen sources. On the basis of this study, a suitable synthetic medium for neomycin production has been developed.     

Cellular Fatty Acid Composition of Selected Pseudomonas Species

The cellular fatty acid composition of 10 reference strains representing eight species of Pseudomonas was determined by gas-liquid chromatography. A variety of acids were detected in these organisms, including branched and straight-chained acids, cyclopropane, and hydroxy acids. Comparison of the presence and relative amounts of these acids among strains was useful for distinguishing various Pseudomonas species.     

Cellular Fatty Acid Composition and Identification of Rumen Bacteria

The fatty acid compositions of 21 pure cultures of rumen bacteria, representing 12 genera and 14 species, were compared as methyl esters. Each organism possessed a consistent and reproducible fatty acid profile. Overlapping similarities and differences in composition did not allow differentiation between families or genera. Although species differentiation was possible, fatty acid composition appeared to be only an aid in the identification of bacteria.     

外源脂肪酸对酒精诱导自絮凝颗粒酵母细胞膜磷脂脂肪酸组成变化的影响

实验将自絮凝颗粒酵母培养于同时添加脂肪酸 (0.6mmol/L)和酒精 (6 %～ 9% ,V/V)条件下以考察其细胞膜磷脂脂肪酸组成的变化。与单独添加棕榈酸相比 ,同时添加酒精引起细胞膜磷脂棕榈酸含量明显增加 ,伴随 9十四碳烯酸、棕榈油酸和油酸含量明显减少 ;与单独添加亚油酸相比 ,同时添加酒精未引起细胞膜磷脂亚油酸含量明显变化 ,但引起油酸含量明显增加 ,伴随 9 十四碳烯酸、棕榈油酸和棕榈酸含量减少 ;与单独添加亚麻酸相比 ,同时添加酒精引起细胞膜磷脂亚麻酸含量减少 ,伴随油酸含量显著增加 ,同时 9 十四碳烯酸、棕榈油酸和棕榈酸含量减少。存活率实验证实 ,上述变化是菌体对酒精刺激的适应性响应 ,因为 ,与培养于仅添加脂肪酸条件下的菌体相比 ,培养于同时添加酒精条件下的菌体耐酒精能力明显提高。研究表明 ,棕榈酸和油酸都可通过加强细胞膜渗透屏障而提高菌体的耐酒精能力 ,这是饱和脂肪酸 (SFA)与不饱和脂肪酸 (UFA)可提高同一菌株耐酒精能力的新的实验现象 ,揭示UFA与SFA在影响酵母菌耐酒精能力的机制上存在共同的作用方式     

Effect of Unsaturated Fatty Acids on the Development of Respiration and on Protein Synthesis in an Unsaturated Fatty Acid Mutant of Saccharomyces cerevisiae

The extent of development of respiratory function induced by aeration of an anaerobically grown unsaturated fatty acid auxotroph of Saccharomyces cerevisiae is determined by the availability, endogenous or externally supplied, of unsaturated fatty acid. The synthesis of mitochondrial and cytoplasmic enzymes during aeration appears to have a similar basis of regulation by available unsaturated fatty acid. Levels of unsaturated fatty acid that permit the synthesis of mitochondrial enzymes also result in a substantial stimulation of cellular protein synthesis.     

Induction of Antibiotic Formation in Streptomyces sp. No. 362 by the Change of Cellular Fatty Acid Spectrum

Microorganisms which require oleic acid for the formation of antibiotics were screened. Streptomyces sp. No. 362, one of the selected organisms, produced antimicrobial substances only when oleic acid, palmitic acid or the high concentration of l-glutamic acid (or l-glutamine) was supplemented to the medium. The cellular fatty acid composition was changed by the supplement of these fatty acids, but not by l-glutamic acid (or l-glutamine). Antibiotic-producing cells had about 4 to 10 times larger amino acid pools, especially l-glutamic acid pool, and hexosamine pools. The ability for l-glutamate uptake of cells grown in the oleic or palmitic acid supplemented medium was markedly enhanced and the efflux of the accumulated l-glutamate was reduced. The antibiotic produced by this strain was identified as one of the streptothricin-group antibiotics and the role of these additives in the antibiotic formation is discussed.     

Association between Plasma Nonesterified Fatty Acids Species and Adipose Tissue Fatty Acid Composition

Fatty acid composition of adipose tissue (AT) is an established long-term biomarker for fatty acid (FA) intake and status, but AT samples are not easily available. Nonesterified FA composition in plasma (pNEFA) may be a good indicator of AT FA composition, because pNEFA are mainly generated by AT lipolysis. We investigated the correlation of 42 pNEFA and subcutaneous as well as visceral AT FA in 27 non-diabetic women with a median BMI of 36 kg/m² (Q_0.25: 25 kg/m²; Q_0.75: 49 kg/m²). Close correlations of pNEFA and AT FA were found for odd-chain FA (15∶0 r = 0.838 and 0.862 for subcutaneous and visceral AT, respectively) and omega-3 FA (22∶6 r = 0.719/0.535), while no significant or low correlations were found for other FA including 18∶1 (r = 0.384/0.325) and 20∶4 (r = 0.386/0.266). Close correlations of pNEFA and AT FA were found for essential fatty acids, like 18∶2 (r = 0.541/0.610) and 20∶5 (r = 0.561/0.543). The lower correlation for some pNEFA species with AT FA indicates that the variation of most pNEFA is significantly affected by other FA sources and flux of FA to tissue, in addition to release from AT. A relevant influence of BMI on the level of correlation was shown for saturated FA. NEFA analysis in fasted plasma can serve as a virtual AT biopsy for some FA, and as a biomarker for intake of dairy products and sea fish.     

Cellular Fatty Acid Composition of Nine Pathovars of Xanthomonas campestris

Cellular fatty acids of 80 strains of Xanthomonas campestris, representing 9 different pathovars, were analyzed by gas-liquid chromatography and mass spectrometry. A total of 48 fatty acids were identified, the most important being the 16:0 (averaging at least 4.5 % of the total), the cis- and trans- 9 16 : 1 (over 14.4 %), and the iso and anteiso 15 : 0 (over 30 %). Other major fatty acids (averaging over 1 % of total) were the saturated 14 : 0 and 15 : 0, the hydroxy-substituted iso 3-OH 11 : 0, 3-OH 12 : 0 and iso 3-OH 13 : 0, and the branch-chained iso 11 : 0, iso 16 : 0, iso 17 : 1, iso 17 : 0 and anteiso 17 : 0. Of 33 minor fatty acids detected and identified, only 7 have been previously reported in the xanthomonads. Significant differences in mean percentages of 5 major fatty acids and 4 (chemical) class totals were detected among pathovars, which statistically segregated into three groups by rank analysis. X. campestris pv. dieffenbachiae was in a group by itself; pvs. campestris, citri (pathotypes A and B), manihotis, phaseoli, pruni and vesicatoria were in a seond group, and pvs. glycines, begonia and citri (pathotype E) were in a third.     

Influence of Exogenous Fatty Acids on Cottonseed Germination

Effects of exogenous free fatty acid application on percentage germination and radicle length were investigated with more chill sensitive (Pima S-4) and less chill sensitive (Pima S-5) cottonseed (Gossypium barbadense L.) at chilling (14°C) and optimal (34°C) temperatures. Of the organic solvents able to solubilize free fatty acids, aqueous solutions as high as 5% dimethyl sulfoxide did not adversely affect germination at 34°C. Palmitic, oleic and linoleic acids (120 μM) were solubilized in 5% DMSO. At 14°C, percent germination of fatty acid treated Pima S-5 significantly increased, while no difference was observed for Pima S-4. Conversely, radicle length of fatty acid treated Pima S-4 significantly increased especially with oleate and linoleate, while no difference was detected for Pima S-5. Fatty acid supplementation influenced neither germination nor radicle length for Pima S-4 and Pima S-5 at 34°C.     

Effect of Endogenous and Exogenous Urate on the Uricase Formation by Streptomyces sp.

Cells of a strain of Streptomyces sp. were incubated with an equivalent quantity of urate, xanthine, 6,8-dihydroxypurine or hypoxanthine in a medium deprived of other nitrogen source. The amount of uricase produced by these cells was shown to differ significantly, increasing in the following order of purine bases added to the medium: urate, xanthine, 6,8-dihydroxypurine and hypoxanthine. Of these was only urate indicated to be the inducer of uricase formation, and the difference in the quantity of uricase produced was found to be based on the duration of enzyme formation. The rate of uricase formation was essentially identical regardless of the purine bases supplied to cells.

Allantoin was accumulated in medium in remarkably different manners depending on the purine bases, which suggested the diversity in the mode of generation of urate in cells. Urate was generated at the slowest rate in the cells incubated with hypoxanthine, although the largest amount of uricase was produced, However, urate supplied to cells at the same rate but from medium failed to support the enzyme formation when the activity increased to a certain level. In order that the same amount of uricase was produced by the cells incubated with the different purine bases, the initial concentration of the purine bases should be raised so that they could remain in medium for the same incubation time.

Intracellular compartmentalization that might segregate endogenous and exogenous urate and might cause the difference in “effeciency” of these urate molecules as the inducer of uricase formation has been discussed.     

Effect of Temperature on the Fatty Acid Composition of Thermus aquaticus

Thermus aquaticus contains four major fatty acids, iso-C(15) (28%), iso-C(16) (9%), normal-C(16) (13%), and iso-C(17) (48%), when grown at 70 C, as determined by gas chromatography and mass spectrometry. Small amounts of iso-C(12), normal-C(12:1), iso-C(13), normal-C(14), iso-C(14), and normal-C(15:1) were also detected. A change in growth temperature (50 to 75 C at 5-C intervals) affects a shift in the proportions of some of the fatty acids. The proportions of the monoenoic and branched-C(17) fatty acids decreased and the proportions of the higher-melting iso-C(16) and normal-C(16) fatty acids increased. Cells grown at 75 C contained 70% more total fatty acids than cells grown at 50 C. The largest increases, in absolute amounts, were in the content of iso-C(16) and normal-C(16) fatty acids, with only a 1.6-fold increase in the major iso-C(15) and iso-C(17) fatty acids. There was a 2.5-fold decrease in normal-C(15:1) and at least a 24-fold decrease in anteiso-C(17), which is present at 50 and 55 C but not at higher temperatures. There was no difference in proportion or amount of fatty acids between exponential and stationary-phase cells grown at 70 C. When cells were grown on glutamate instead of yeast-extract and tryptone at 70 C, the total fatty acid content remained constant, but there was an increase in the proportions of iso-C(16) and normal-C(16) fatty acids concomitant with a decrease in the proportions of the iso-C(15) and iso-C(17) fatty acids.     

Effect of Substrate on the Fatty Acid Composition of Hydrocarbon-utilizing Microorganisms

The fatty acid pattern in three hydrocarbon-utilizing bacteria during growth on various substrates was examined. The predominant fatty acids in acetate-grown cells were C(16), C(16:1), C(18:1), and Br-C(19) and the major fatty acids in propane-grown cells were C(15), C(17), C(17:1), C(18:1), and Br-C(18). When one organism (Mycobacterium sp. strain OFS) was grown on the n-alkanes from C(13) to C(17), the major fatty acid in the cells was of the same chain length as the substrate. Studies on the incorporation of acetate into the cellular fatty acids of microorganisms growing on C(15) and C(17)n-alkanes suggest that the oxidative products of the substrate are incorporated into the cellular fatty acids without degradation to acetate.