Fatty acid composition of Mesorhizobium huakuii lipopolysaccharides. Identification of 27-oxooctacosanoic acid

Lipopolysaccharides of three Mesorhizobium huakuii strains carried a number of amide-linked 3-hydroxylated fatty acids including: 3-OH-12:0, 3-OH-i-13:0, 3-OH-20:0, 3-OH-i-21:0, 3-OH-22:0, 3-OH-23:0 and unsaturated 3-OH-22:1. The first three of the above mentioned acids are the main amide-linked fatty acids in the LPS preparations. The main ester-bound fatty acids comprise 16:0, i-17:0, 18:0, 20:0 and 27-OH-28:0. Among minor constituents of lipid A 25-OH-26:0 and 29-OH-30:0 together with some non-polar fatty acids were found. Additionally, the presence of 4-oxo-20:0, 4-oxo-i-21:0 and 4-oxo-22:0 amide-bound fatty acids as well as the 27-oxo-28:0 ester-linked fatty acid were proved. To our knowledge oxo fatty acids are rare constituents of lipopolysaccharides and 27-oxo-28:0 was found for the first time in the LPS preparations from members of Rhizobiaceae.     

Distribution and phylogenetic significance of 27-hydroxy-octacosanoic acid in lipopolysaccharides from bacteria belonging to the alpha-2 subgroup of Proteobacteria.

Lipopolysaccharides (LPSs) from a number of bacteria belonging to the alpha-2 subgroup of the class Proteobacteria were screened for the presence of 27-hydroxy-octacosanoic acid (27-OH-28:0). With few exceptions, most of the bacteria contained 27-OH-28:0 in their lipid A fractions. In addition, some of the bacteria contained other n-2-hydroxylated long-chain fatty acids hitherto not reported. The distribution of 27-OH-28:0 was restricted to the alpha-2 subgroup. LPSs from members of the other subgroups (the beta and gamma subgroups), including some well-characterized enterobacterial LPSs, were devoid of 27-OH-28:0. Our results indicate that the presence of n-2-hydroxylated long-chain fatty acids in LPSs might be used as a chemophylogenetical marker.     

Lipopolysaccharides from Mesorhizobium huakuii and Mesorhizobium ciceri: chemical and immunological comparative data

Lipopolysaccharides of two Mesorhizobium species of different host specificity were compared: M. huakuii and M. ciceri. M. huakuii sp. was represented by five strains with special consideration of M. huakuii IFO 15243(T). SDS/PAGE profiles revealed that all M. huakuii LPS preparations contained low molecular mass fractions (LPS-II) of the same molecular size. All of lipopolysaccharides contained high molecular mass fractions (LPS-I). However, the high molecular mass fraction from each strain possessed an individual molecular size distribution pattern. The crossreactivity of blotted lipopolysaccharides with rabbit polyclonal antibodies against Mesorhizobium huakuii IFO 15243(T) whole bacteria indicated the presence of common epitope(s) within the investigated Mesorhizobium huakuii strains. Moreover, LPS from M. huakuii S52 also reacted with anti M. ciceri HAMBI 1750 serum showing that there are epitopes common for different mesorhizobial species. LPS isolated from Mesorhizobium huakuii strain IFO 15243(T) contained neutral sugars: L-6-deoxytalose, L-rhamnose, D-galactose and D-glucose, aminosugars:D-quinovosamine, D-glucosamine, D-2,3-diamino-2,3-dideoxyglucose and D-galacturonic and D-glucuronic acids. In the LPS preparation, fatty acids typical for Mesorhizobium strains were detected. 3-Hydroxydodecanoic, 3-hydroxy-iso-tridecanoic, 3-hydroxyeicosanoic, 3-hydroxyheneicosanoic and 3-hydroxydocosenoic acids were the major amide linked fatty acids, while iso -heptadecanoic, eicosanoic, docosenoic, as well as 27-hydroxyoctacosanoic and 27-oxooctacosanoic acids were the dominant ester linked fatty residues.     

Long-chain α-hydroxy-(ω-1)-oxo fatty acids and α-hydroxy-1,ω-dioic fatty acids are cell wall constituents of Legionella (L. jordanis, L. maceachernii and L. micdadei)

Abstract Four long-chain fatty acids, 2-hydroxy-27-oxo-octacosanoic acid ( n 28:0(2-OH,27-oxo)), 2-hydroxy-29-oxo-triacontanoic acid ( n 30:0(2-OH,29-oxo)), 2-hydroxy-heptacosane-1,27-dioic acid (27:0(2-OH)-dioic) and 2-hydroxy-nonacosane-1,29-dioic acid (29:0(2-OH)-dioic) were identified by GLC-MS analysis in the phenol-chloroform-petroleum ether (PCP) extracts of Legionella jordanis, L. maceachernii and L. micdadei indicating that they are constituents of lipopolysaccharide. Moreover, five long-chain fatty acids (28:0(27-OH), 28:0(27-oxo), 30:0(29-oxo), 27:0-dioic and 29:0-dioic) previously identified in L. pneumophila (Moll, H. et al., FEMS Microbiol. Lett., 97 (1992), 1–6) were also found in these species. This is to our knowledge the first report on the existence of long chain 2-hydroxylated (ω-1)-oxo fatty acids and 2-hydroxylated 1,ω-dioic fatty acids.     

Occurrence of lipid A variants with 27-hydroxyoctacosanoic acid in lipopolysaccharides from members of the family Rhizobiaceae.

Lipopolysaccharides (LPSs) isolated from several strains of Rhizobium, Bradyrhizobium, Agrobacterium, and Azorhizobium were screened for the presence of 27-hydroxyoctacosanoic acid. The LPSs from all strains, with the exception of Azorhizobium caulinodans, contained various amounts of this long-chain hydroxy fatty acid in the lipid A fractions. Analysis of the lipid A sugars revealed three types of backbones: those containing glucosamine (as found in Rhizobium meliloti and Rhizobium fredii), those containing glucosamine and galacturonic acid (as found in Rhizobium leguminosarum bv. phaseoli, trifolii, and viciae), and those containing 2,3-diamino-2,3-dideoxyglucose either alone or in combination with glucosamine (as found in Bradyrhizobium japonicum and Bradyrhizobium sp. [Lupinus] strain DSM 30140). The distribution of 27-hydroxyoctacosanoic acid as well as analysis of lipid A backbone sugars revealed the taxonomic relatedness of various strains of the Rhizobiaceae.     

Intraspecies variability of cellular fatty acids among soil and intestinal strains of Desulfovibrio desulfuricans.

A comparison of cellular fatty acid profiles of Desulfovibrio desulfuricans DSM 642 and 14 wild strains of this species, isolated from two completely different environments, soil and the human intestine, was carried out. All the D. desulfuricans strains grown on lactate and sulfate indicated the presence of considerable amounts of i-C15:0, i-C17:1 and C16:0. Although differences in the quantities of individual fatty acids present in each strain were clear in the group of soil strains (similarity, 67.6%), in contrast to almost identical fatty acid patterns (similarity, near 100%) in the intestinal strains, the results were variable within the limits acceptable for species demonstration. The higher similarity of the fatty acid profiles of intestinal strains may be a result of the similarity of biocenoses in the human digestive tract. The coefficients of variability of i-C17:1 and i-C15:0 (the major branched-chain fatty acids), as well as clustering of the investigated strains compared with strains described in the literature after plotting percentages of i-C17:1 fatty acid against i-C15:0 fatty acid, confirmed a certain heterogeneity of cellular fatty acid profiles within the group of soil strains, in contrast to almost ideal homogeneity within the group of intestinal isolates. Intestinal strains contained a higher ratio of saturated to unsaturated fatty acids (2.2 +/- 0.14) than did soil strains (1.6 +/- 0.2; in one case, 2.7). We propose that intestinal D. desulfovibrio bacteria should be assumed to be a highly homogeneous group and should be represented by the strain D. desulfuricans subsp. intestinus in collections of microbial cultures.     

Chemical characterization of effective and ineffective strains of Rhizobium leguminosarum bv. viciae

Chemical composition of lipopolysaccharide (LPS) isolated from an effective (97) and ineffective (87) strains of R. l. viciae has been determined. LPS preparations from the two strains contained: glucose, galactose, mannose, fucose, arabinose, heptose, glucosamine, galactosamine, quinovosamine, and 3-N-methyl-3,6-dideoxyhexose, as well as glucuronic, galacturonic and 3-deoxyoctulosonic acid. The following fatty acids were identified: 3-OH 14:0, 3-OH 15:0, 3-OH 16:0, 3-OH 18:0 and 27-OH 28:0. The ratio of 3-OH 14:0 to other major fatty acids in LPS 87 was higher that in LPS 97. SDS/PAGE profiles of LPS indicated that, in lipopolysaccharides, relative content of S form LPS I to that of lower molecular mass (LPS II) was much higher in the effective strain 97 than in 87. All types of polysaccharides exo-, capsular-, lipo, (EPS, CPS, LPS, respectively) examined possessed the ability to bind faba bean lectin. The degree of affinity of the host lectin to LPS 87 was half that to LPS 97. Fatty acids (FA) composition from bacteroids and peribacteroid membrane (PBM) was determined. Palmitic, stearic and hexadecenoic acids were common components found in both strains. There was a high content of unsaturated fatty acids in bacteroids as well as in PBM lipids. The unsaturation index in the PBM formed by strain 87 was lower than in the case of strain 97. Higher ratio of 16:0 to 18:1 fatty acids was characteristic for PMB of the ineffective strain.     

Cell wall constituents of Microcystis sp. PCC 7806

Cell walls of Microcystis sp. PCC 7806 were purified from cell homogenates by sucrose density centrifugation and Triton X-100 extraction. The outer membrane contained carotenoids, two major peptidoglycan-associated proteins (Mr 49,000 and 52,000), and lipopolysaccharide (LPS) as indicated by the presence of 3-hydroxy fatty acids (3-OH-14:0, 3-OH-16:0, 3-OH-18:0), 4-oxo-18:0 fatty acid, and GlcN as lipid A components in addition to rare O-methyl sugars (2-O-methyl-6-deoxyhexoses I and II). The peptidoglycan (A1 gamma-type) was found to be covalently linked to a wall polysaccharide composed of GlcN, ManN, Man, Glc, and phosphate.     

LIPID CLASS DISTRIBUTION OF HIGHLY UNSATURATED LONG CHAIN FATTY ACIDS IN MARINE DINOFLAGELLATES

The very long chain highly unsaturated C₂₈ fatty acids, octacosaheptaenoic [28:7(n-6)] and octacosaoctaenoic acid [28:8(n-3)], were found to be associated with phospholipids, obtained by fractionation of total lipid extracts into distinct lipid classes, in 4 and 6, respectively, of 16 examined dinoflagellates. An interfraction comparison of fatty acids associated with phospholipids and glycolipids has also shown that the phospholipid fractions contained the majority (over 75% in 12 of 16 strains) of docosahexaenoic acid [22:6(n-3)] and traces of tetracosanoic acid (24:0). By contrast, the highly unsaturated C₁₈ fatty acids octadecatetraenoic [18:4(n-3)] and octadecapentaenoic acid [18:5(n-3)] were primarily recovered from a chloroplast-associated glycolipid fraction comprised of monogalactosyldiacylglycerol, digalactosyldiacylglycerol, and sulfoquinovosyldiacylglycerol. In 12 of 16 strains, an interfraction comparison showed that over 90% of 18:5(n-3) was found to be associated with glycolipids. These findings indicate that the C₂₈ fatty acids are located and probably synthesized in the cytoplasm or in an organelle other than the chloroplast, possibly with 22:6(n-3) and 24:0 as precursors, whereas the C₁₈ fatty acids 18:4(n-3) and 18:5(n-3) are glycolipid constituents apparently synthesized within the chloroplast. The function(s) of these C₂₈ fatty acids as components of phospholipids in cellular membranes is currently unknown.     

Analysis of cellular fatty acids in Orientia tsutsugamushi as taxonomic markers

Six Orientia strains including 3 prototype strains such as Gilliam, Karp, and Kato, and 3 strains (Boryong, Pajoo, and Yongworl) isolated in Korea, were studied for the profiles of their cellular fatty acids. All tested strains contained octadecenoic acid C (18: 1) omega 9 c(57.3 +/- 3.5%), octadecanoic acid C (18: 0) (15.3 +/- 1.5%), and hexadecanoic acid C (16: 0) (12.7 +/- 1.7%) as major components; however, interestingly, eicosenoic acid C (20: 1) omega 9 c(2.6 +/- 0.6%) was found in all strains except the Yongworl strain. Furthermore none of the strains contained 3-hydroxy fatty acids. The ratios of total saturated fatty acid (SFA) to total unsaturated fatty acid (UFA) were within the range of 0.34 to 0.54. These results showed that the cellular fatty acid profile should provide more reliable information for the identification of these bacteria.     

Chemical analysis of Azospirillum lipopolysaccharides

Lipopolysaccharides (LPS) were extracted by hot phenol-water from five strains each of Azospirillum lipoferum and Azospirillum brasilense. Rhamnose, glucose, glucosamine and 3-deoxy-d-mannooctulosonic acid were comon sugar constituents of all LPS preparations. 2-O-Mefucose, 3-O-Me-fucose, 3-O-Me-rhamnose and 2-O-Megalactose were found in LPSs of some A. brasilense strains. Fatty acid spectra from all LPSs studied were almost identical with predominance of 3-hydroxymyristic and 3-hydroxypalmitic acids. 3-Hydroxypalmitic acid was the only amide-linked fatty acid. Lipopolysaccharides isolated from A. brasilense showed higher heterogeneity in sugar composition than those from A. lipoferum.Abbreviations glc gas liquid chromatography - ms mass spectrometry - LPS lipopolysaccharide - dOclA 3-deoxy-d-mannooctulosonic acid - 3-OH-16:0 3-hydroxypalmitic acid - nir^- nitrite reductase negative - nir⁺ nitrite reductase positive     

Differential effects of 2-oxo acids on pyruvate utilization and fatty acid synthesis in rat brain

1. The effects of 2-oxo-4-methylpentanoate, 2-oxo-3-methylbutanoate and 2-oxo-3-methylpentanoate on the activity of pyruvate dehydrogenase (EC 1.2.4.1), citrate synthase (EC 4.1.3.7), acetyl-CoA carboxylase, (EC 6.4.1.2) and fatty acid synthetase derived from the brains of 14-day-old rats were investigated. 2. The pyruvate dehydrogenase enzyme activity was competitively inhibited by 2-oxo-3-methylbutanoate with respect to pyruvate with a K(i) of 2.04mm but was unaffected by 2-oxo-4-methylpentanoate or 2-oxo-3-methylpentanoate. 3. The citrate synthase activity was inhibited competitively (with respect to acetyl-CoA) by 2-oxo-4-methylpentanoate (K(i)~7.2mm) and 2-oxo-3-methylbutanoate (K(i)~14.9mm) but not by 2-oxo-3-methylpentanoate. 4. The acetyl-CoA carboxylase activity was not inhibited significantly by any of the 2-oxo acids investigated. 5. The fatty acid synthetase activity was competitively inhibited (with respect to acetyl-CoA) by 2-oxo-4-methylpentanoate (K(i)~930mum) and 2-oxo-3-methylpentanoate (K(i)~3.45mm) but not by 2-oxo-3-methylbutanoate. 6. Preliminary experiments indicate that 2-oxo-4-methylpentanoate and 2-oxo-3-phenylpropionate (phenylpyruvate) significantly inhibit the ability of intact brain mitochondria from 14-day-old rats to oxidize pyruvate. 7. The results are discussed with reference to phenylketonuria and maple-syrup-urine disease. A biochemical mechanism is proposed to explain the characteristics of these diseases.     

Ketonic bile acids in urine of infants during the neonatal period

Ketonic bile acids have been found to be quantitatively important in urine of healthy infants during the neonatal period. In order to determine their structures, the bile acids in urine from 11 healthy infants were analyzed by gas-liquid chromatography-mass spectrometry (GLC-MS) and three samples with particularly high levels of ketonic bile acids were selected for detailed studies by ion exchange chromatography, fast atom bombardment mass spectrometry, microchemical reactions, and GLC-MS. The major ketonic bile acid was identified as 7 alpha, 12 alpha-dihydroxy-3-oxo-5 beta-chol-1-enoic acid, not previously described as a naturally occurring bile acid. The positional isomer 7 alpha, 12 alpha-dihydroxy-3-oxo-4-cholenoic acid, recently described as a major urinary bile acid in infants with severe liver diseases, was also excreted by most infants. Three acids related to cholic acid were identified: 7 alpha, 12 alpha-dihydroxy-3-oxo-, 3 alpha, 12 alpha-dihydroxy-7-oxo-, and 3 alpha, 7 alpha-dihydroxy-12-oxo-5 beta-cholanoic acids. Five bile acids having one oxo and three hydroxy groups were also present. Based on mass spectra and biological considerations two of these were tentatively given the structures 1 beta, 7 alpha, 12 alpha-trihydroxy-3-oxo- and 1 beta, 3 alpha, 12 alpha-trihydroxy-7-oxo-5 beta-cholanoic acids. Some of the others had a hydroxy group at C-4 or C-2. The levels of ketonic bile acids were higher on the third than on the first day of life, and lower after 1 month. The formation and excretion especially of 3-oxo bile acids is proposed to result from changes of the redox state in the liver in connection with birth.     

Structural analyses of the lipopolysaccharides from Chlamydophila psittaci strain 6BC and Chlamydophila pneumoniae strain Kajaani 6.

Lipopolysaccharides (LPSs) of Chlamydophila psittaci 6BC and Chlamydophila pneumoniae Kajaani 6 contain 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo), GlcN, organic bound phosphate, and fatty acids in the molar ratios of approximately 3:2:2.2:4.8 and approximately 2.9:2:2.1:4.9, respectively. The LPSs were immunoreactive with a monoclonal antibody against a family-specific epitope of chlamydial LPS. This finding, together with methylation analyses of both LPSs and MALDI-TOF MS experiments on de-O-, and de-O,N-acylated LPSs, indicate the presence of a Kdo trisaccharide proximal to lipid A having a structure alpha-Kdo-(2-->8)-alpha-Kdo-(2-->4)-alpha-Kdo, which appears to be the main component of the core region in the native chlamydial LPSs. In the de-O-acylated LPSs from Chl. psittaci 6BC and Chl. pneumoniae Kajaani 6, two major molecular species are present that differ in distribution of amide-bound hydroxy fatty acids over both GlcN. It appears that either two (R)-3-hydroxy-18-methylicosanoic acids or one (R)-3-hydroxy-18-methylicosanoic acid and one (R)-3-hydroxyicosanoic acid are attached to the GlcN residues. In contrast, the de-O-acylated LPS of Chl. psittaci PK 5082 contains one major molecular species that has two (R)-3-hydroxyicosanoic acid residues attached to two GlcN residues.     

Analysis of phospholipids and ornithine-containing lipids from Mesorhizobium spp

Polar lipid compositions of seven strains belonging to the four species of the Mesorhizobium genus were described. The lipid patterns of Mesorhizobium strains were very similar. Only quantitative differences were observed. Diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), phosphatidylethanolamine (PE), and phosphatidylcholine (PC) were found to be the major phospholipids of the analysed bacteria. In addition, two methylated derivatives of PE were observed: phosphatidyl-N,N-dimethylethanolamine (DMPE) and phosphatidyl-N-monomethylethanolamine (MMPE). Polar head groups of those phospholipids were predominately acylated with lactobacillic (19:0 cyclopropane) acid. Ornithine-containing lipid (OL) was also identified. 3-hydroxy fatty acids found in the lipid preparations were derived exclusively from the ornithine lipid. 3-hydroxylactobacillic was the main acyl residue amide linked to the ornithine.     

Chemotaxonomic differentiation of coryneform bacteria isolated from biofilters.

Coryneform bacteria that were isolated from biofilters which are used for waste gas treatment of animal-rendering plant emissions were differentiated and partially identified by using chemotaxonomic methods. On the basis of the results of a numerical analysis of whole-cell fatty acid profiles, 79 isolates were divided into two major groups; the members of the first group contained saturated and monounsaturated fatty acids, whereas the members of the second group were characterized by iso- and anteiso-branched fatty acids. Division into subclusters was based mainly on quantitative differences in fatty acid composition and was confirmed by the results obtained for additional chemical markers (e.g., respiratory quinones, mycolic acids, polar lipids, cell wall amino acids, and whole-cell sugar patterns). By combining the results obtained for chemotaxonomic analyses that were performed for strains containing saturated and monounsaturated fatty acids, we were able to identify the genus Corynebacterium (two Corynebacterium species were differentiated on the basis of the occurrence of tuberculostearic acid), the genus Gordona, and the genus Mycobacterium. Among the strains that produced iso-anteiso fatty acid patterns, one subgroup was affiliated with the "nicotianae" group of the genus Arthrobacter; however, some strains contained a new combination of chemical markers. Peptidoglycan type A4 alpha, L-Lys-Gly-L-Glu was combined with menaquinones MK-7 and MK-8, whereas peptidoglycan type A4 alpha, L-Lys-L-Glu occurred together with MK-8 and MK-9. The second subgroup was characterized by a new type B peptidoglycan and MK-11, as well as small amounts of MK-12. Differentiation that was based first on chemotaxonomy and second on physiology gave reliable results. Thus, coryneform strains with new characteristics were isolated from biofilters.     

Analysis of fatty acids from lipopolysaccharides of Bacteroides thetaiotaomicron and Bacteroides fragilis]

The aim of this study was to determine and to compare fatty acids occurring in lipopolysaccharides (LPS) isolated from B. thetaiotaomicron and B. fragilis strains of different origin. Lipopolysaccharides of three B. thetaiotaomicron strains and four B. fragilis strains were isolated by phenol-water extraction according to the procedure of Westphal and Jann (1965). Water-phase LPS fractions were then treated with nucleases and purified by ultracentrifugation as described by Gmeiner (1975). Fatty acid methyl esters, obtained by methanolysis of LPS, were analysed in gas-liquid chromatography combined with mass spectrometry (GLC-MS). Trimethylsilylated hydroxyl groups of fatty acid methyl esters were identified with GLC-MS using a method of selective ion monitoring (SIM). Lipopolysaccharides of B. thetaiotaomicron and B. fragilis strains contained long-chain (15-18 carbon atoms) fatty acids. The broad spectrum of simple long-chain and branched-chain fatty acids as well as 3-hydroxy fatty acids were detected. The main fatty acid of analyzed bacterial species was 3-hydroxy-hexadecanoic acid (3OH C16:0). Several 3-hydroxy fatty acids were detected in LPS of examined strains. Fatty acids occurring in LPS of B. thetaiotaomicron and B. fragilis strains appeared to be qualitatively similar. Quantitative differences in fatty acids composition of lipopolysaccharides isolated from strains of different origin were observed.     

Long chain oxoaldehydes and oxoalcohols from esters as major constituents of the surface lipids of Manduca sexta pupae in diapause

Lipid constitutents of diapausing pupae of the tobacco hornworm, Manduca sexta (L), were identified by thin layer and gas-liquid chromatography, IR spectroscopy, and gas-liquid chromatography-mass spectrometry. Surface wax was a mixture of long chain polar compounds: oxoalcohol esters, oxoaldehydes, primary alcohol esters, and oxoalcohols, as listed in descending order of abundance. The distribution of the alcohols and aldehydes was C28 (75-85%), C27 (5%), and C26 (10-15%). The C26 compounds were largely 11-oxo isomers, but the C28 compounds consisted of similar amounts of 11- and 12-oxo isomers. The identities of the oxoaldehydes were confirmed by selective and complete NaBH4 reductions to yield oxoalcohols and diols, respectively. Mass spectral interpretations were verified with mass spectra of the oxoaldehyde, oxoalcohol, and diol synthesized from 12-hydroxystearic acid. Reduction of the total lipids with NaBH4 and hydrolysis of the product with ethanolic KOH gave oxoalcohols (85%), primary alcohols (8%), and oxoacids (5%); 30-40% of the oxoalcohols were derived from NaBH4-reduced oxoaldehydes, 5-10% were from free oxoalcohols, and 50% were from esters. Primary alcohols only existed as esters. Large quantities of fatty oxoalcohols relative to fatty oxoacids in the saponified mixture suggested the presence of esters other than those composed of long chain acids and alcohols. Oxoacids appeared to be mainly oxidation products of the oxoaldehydes.     

Polar lipid and fatty acid profiles – Re-vitalizing old approaches as a modern tool for the classification of mycoplasmas?

A set of 20 Mollicutes strains representing different lines of descent, including the type species of the genus Mycoplasma, Mycoplasma mycoides, Acholeplasma laidlawii and a strain of Mesoplasma, were subjected to polar lipid and fatty acid analyses in order to evaluate their suitability for classification purposes within members of this group. Complex polar lipid and fatty acid profiles were detected for each examined strain. All strains contained the polar lipids phosphocholine-6'-alpha-glucopyranosyl-(1'-3)-1, 2-diacyl-glycerol (MfGL-I), 1-O-alkyl/alkenyl-2-O-acyl-glycero-3-phosphocholine (MfEL), sphingomyelin (SphM), 1-O-alkyl/alkenyl-glycero-3-phosphocholine (lysoMfEL), the unknown aminophospholipid APL1 and the cholesterol Chol2. A total of 19 strains revealed the presence of phosphatidylethanolamine (PE) and/or phosphatidylglycerol (PG), and the presence of diphosphatidylglycerol (DPG) was detected in 13 strains. The unknown aminolipid AL1 was found in the extracts of 17 strains. Unbranched saturated and unsaturated compounds predominated in the fatty acid profiles. Major fatty acids were usually C16:0, C18:0, C18:1 omega9c and 'Summed feature 5' (C18:2 omega6, 9c/C18:0 anteiso). Our results demonstrated that members of the M. mycoides cluster showed rather homogenous polar lipid and fatty acid profiles. In contrast, each of the other strains was characterized by a unique polar lipid profile and significant quantitative differences in the presence of certain fatty acids. These results indicate that analyses of both polar lipid and fatty acid profiles could be a useful tool for classification of mycoplasmas.     

Chemotaxonomy of heterocystous cyanobacteria using FAME profiling as species markers

The fatty acid methyl ester (FAME) analysis of the 12 heterocystous cyanobacterial strains showed different fatty acid profiling based on the presence/absence and the percentage of 13 different types of fatty acids. The major fatty acids viz. palmitic acid (16:0), hexadecadienoic acid (16:2), stearic acid (18:0), oleic acid (18:1), linoleic (18:2), and linolenic acid (18:3) were present among all the strains except Cylindrospermum musicola where oleic acid (18:1) was absent. All the strains showed high levels of polyunsaturated fatty acid (PUFAs; 41-68.35%) followed by saturated fatty acid (SAFAs; 1.82-40.66%) and monounsaturated fatty acid (0.85-24.98%). Highest percentage of PUFAs and essential fatty acid (linolenic acid; 18:3) was reported in Scytonema bohnerii which can be used as fatty acid supplement in medical and biotechnological purpose. The cluster analysis based on FAME profiling suggests the presence of two distinct clusters with Euclidean distance ranging from 0 to 25. S. bohnerii of cluster I was distantly related to the other strains of cluster II. The genotypes of cluster II were further divided into two subclusters, i.e., IIa with C. musicola showing great divergence with the other genotypes of IIb which was further subdivided into two groups. Subsubcluster IIb(1) was represented by a genotype, Anabaena sp. whereas subsubcluster IIb(2) was distinguished by two groups, i.e., one group having significant similarity among their three genotypes showed distant relation with the other group having closely related six genotypes. To test the validity of the fatty acid profiles as a marker, cluster analysis has also been generated on the basis of morphological attributes. Our results suggest that FAME profiling might be used as species markers in the study of polyphasic approach based taxonomy and phylogenetic relationship.