Structures of Hexadecenoic Acid and Octadecenoic Acid in Lipomyces starkeyi

Taxonomic and some other properties of a yeast strain, Candida sp. 36, which characteristically assimilates n-alkanes, were described. Identification of coenzyme Q, NMR spectroscopy of cell wall polysaccharides, determination of G+C content of DNA and some DNA-DNA hybridization experiments were carried out, in addition to the morphological and physiological observations. All the data were consistent with the suggestion that Candida cloacae Komagata, Nakase and Katsuya and Candida subtropicalis Nakase, Fukazawa and Tsuchiya are the synonyms of Candida maltosa Komagata, Nakase and Katsuya. Candida sp. 36 was identified as C. maltosa, too. The yeast was found to grow most abundantly on n-hexadecane and on n-octadecane in the presence of biotin.     

Studies on the Enzymatic Production of l-Aspartic Acid from Maleic Acid

The enzymatic production of l-aspartic acid from maleic acid with cell suspensions of Alcaligenes faecalis 5-24, isolated from solid by the authors, was investigated.

The optimum conditions of this reaction and some cultural conditions which influenced on the ability of the cells to catalyze the above reaction were mainly studied.

The cells grown on maleic acid as a sole source of carbon showed exclusively the strong ability. The cells grown on a carbon source other than maleic acid showed no activity of this reaction.

It was concluded that an inducibles enzyme whose formation was stimulated by the presence of maleic acid might be involved in the reaction for the production of l-aspartic acid from maleic acid.

It was found that malonic acid was replaceable for maleic acid which played an inductive role for the formation of the enzyme system concerned with the reaction of l-aspartic acid production from maleic acid.

The cells grown in the medium containing malonic acid showed a stronger activity of the above reaction than the cells grown on maleic acid. The induction effect of malonic acid was remarkable when the organism was cultured in an acid medium. Whereas, consumption of C¹⁴-malonic acid in the medium by the organism was not observed at all in any pH milieu even where the formation of the enzyme system essential for the reaction was fully conducted. It indicated that malonic acid penetrated preferentially in acid milieu into the cells was a non-metabolic inducer like thiomethyl-β-d-galactoside in β-galactosidase system and that permeability barrier might exist in the organism.

The formation of cis-trans isomerase which catalyzed the conversion of maleic acid to fumaric acid was much stimulated by the addition of either malonic acid or maleic acid. From these results, it was concluded that l-aspartic acid was produced from maleic acid and ammonium ion by both actions of the inducible cis-trans isomerase and the constitutive aspartase.     

A Mutant of Arabidopsis Deficient in the Elongation of Palmitic Acid

The overall fatty acid composition of leaf lipids in a mutant of Arabidopsis thaliana was characterized by an increased level of 16:0 and a concomitant decrease of 18-carbon fatty acids as a consequence of a single recessive nuclear mutation at the fab1 locus. Quantitative analysis of the fatty acid composition of individual lipids established that lipids synthesized by both the prokaryotic and eukaryotic pathways were affected by the mutation. Direct enzyme assays demonstrated that the mutant plants were deficient in the activity of 3-ketoacyl-acyl carrier protein synthase II; therefore, it is inferred that fab1 may encode this enzyme. Labeling experiments with [14C]acetate and lipase positional analysis indicated that the mutation results in a small shift in the partitioning of lipid synthesis between the prokaryotic and eukaryotic pathways. Synthesis of chloroplast lipids by the prokaryotic pathway was increased with a corresponding reduction in the eukaryotic pathway.     

Hexadecenoic Fatty Acid Isomers in Human Blood Lipids and Their Relevance for the Interpretation of Lipidomic Profiles

Monounsaturated fatty acids (MUFA) are emerging health biomarkers, and in particular the ratio between palmitoleic acid (9cis-16:1) and palmitic acid (16:0) affords the delta-9 desaturase index that is increased in obesity. Recently, other positional and geometrical MUFA isomers belonging to the hexadecenoic family (C16 MUFA) were found in circulating lipids, such as sapienic acid (6cis-16:1), palmitelaidic acid (9trans-16:1) and 6trans-16:1. In this work we report: i) the identification of sapienic acid as component of human erythrocyte membrane phospholipids with significant increase in morbidly obese patients (n = 50) compared with age-matched lean controls (n = 50); and ii) the first comparison of erythrocyte membrane phospholipids (PL) and plasma cholesteryl esters (CE) in morbidly obese patients highlighting that some of their fatty acid levels have opposite trends: increases of both palmitic and sapienic acids with the decrease of linoleic acid (9cis,12cis-18:2, omega-6) in red blood cell (RBC) membrane PL were reversed in plasma CE, whereas the increase of palmitoleic acid was similar in both lipid species. Consequentially, desaturase enzymatic indexes gave different results, depending on the lipid class used for the fatty acid content. The fatty acid profile of morbidly obese subjects also showed significant increases of stearic acid (C18:0) and C20 omega-6, as well as decreases of oleic acid (9cis-18:1) and docosahexaenoic acid (C22:6 omega-3) as compared with lean healthy controls. Trans monounsaturated and polyunsaturated fatty acids were also measured and found significantly increased in both lipid classes of morbidly obese subjects. These results highlight the C16 MUFA isomers as emerging metabolic marker provided that the assignment of the double bond position and geometry is correctly performed, thus identifying the corresponding lipidomic pathway. Since RBC membrane PL and plasma CE have different fatty acid trends, caution must also be used in the choice of lipid species for the interpretation of lipidomic profiles.     

Enzymatic Formation of Sphingenine from Ketosphinganine in Rat Liver Particulates

Mutants of Klebsiella aerogenes that synthesized tyramine oxidase and arylsulfatase constitutively were isolated. The properties of four of seven constitutive mutants isolated were unstable, segregating spontaneously to the parental type at high frequency. Some of these segregants also lost arylsulfatase (AtsA^?) or tyramine oxidase (TynA^?) spontaneously. These unstable constitutive mutations were shown by genetic analysis to involve mutations of tynP and tynR, and transductants receiving the tynP gene were also unstable. These results showed that the instability was due to unstable tynP gene, which may be the promoter region for tyramine oxidase.     

Dependence of Arbuscular-Mycorrhizal Fungi on Their Plant Host for Palmitic Acid Synthesis

Lipids are the major form of carbon storage in arbuscular-mycorrhizal fungi. We studied fatty acid synthesis by Glomus intraradices and Gigaspora rosea. [¹⁴C]Acetate and [¹⁴C]sucrose were incorporated into a synthetic culture medium to test fatty acid synthetic ability in germinating spores (G. intraradices and G. rosea), mycorrhized carrot roots, and extraradical fungal mycelium (G. intraradices). Germinating spores and extraradical hyphae could not synthesize 16-carbon fatty acids but could elongate and desaturate fatty acids already present. The growth stimulation of germinating spores by root exudates did not stimulate fatty acid synthesis. 16-Carbon fatty acids (16:0 and 16:1) were synthesized only by the fungi in the mycorrhized roots. Our data strongly suggest that the fatty acid synthase activity of arbuscular-mycorrhizal fungi is expressed exclusively in the intraradical mycelium and indicate that fatty acid metabolism may play a major role in the obligate biotrophism of arbuscular-mycorrhizal fungi.     

Assessment of Fetal Lung Maturity by Estimation of Amniotic Fluid Palmitic Acid

The measurement of palmitic acid in amniotic fluid has been shown to be a rapid means of assessing the lecithin concentration. The level of palmitic acid increases quickly when the fetal lung matures at about 35 weeks'' gestation, and the level in amniotic fluid obtained 24 hours or less before delivery clearly distinguishes which infants are likely to develop respiratory distress syndrome and which are mature.     

Acid and Enzymatic Hydrolysis of an Acidic Polysaccharide from Soy Sauce

Mild acid hydrolysis of an acidic polysaccharide (APS-I) from soy sauce resulted in a degraded polysaccharide (DPS), the mixture of neutral sugar, D-galacturonic acid, its α-1,4-linked homologous di- and trisaccharides, and acidic oligosaccharides containing residues of D-galacturonic acid and L-rhamnose. Besides the above-mentioned sugars, an aldobiouronic acid containing D-xylose moiety was also yielded in the enzymatic hydrolysates with a crude polysaccharidase preparation. However, only a β-l, 4-galactobiose was isolated from the lower molecular fraction of enzymatic digest of APS-I with a typical hemicellulase preparation. DPS containing 83% of D-galacturonic acid was able to be degraded by endo-polygalacturonase, but APS-I was not because of its highly was discussed on the basis of these results, periodate oxidation study.     

棕榈酸促进肝癌细胞侵袭转移的分子机制

目的:探讨棕榈酸(Palmiticacid,PA)对人肝癌细胞系SMMC-7721侵袭转移能力的影响,并通过检测肝癌细胞系中CD147-MMPs信号通路在PA影响下的变化,初探PA影响肝癌细胞侵袭转移的分子机制。方法:PA(0、20、50、100μM)作用SMMC-7721细胞后(8、16、24h),MTT法检测细胞增殖,划痕及Transwell实验评价细胞迁移侵袭能力,Western-blot及real-time PCR检测CD147蛋白及其mRNA的水平,ELISA检测基质金属蛋白酶(MMP-2,MMP-9)的水平。结果:与对照组相比,PA作用SMMC-7721细胞后,细胞存活率无显著差异(P0.05);细胞迁移和侵袭能力显著增高(P0.05);CD147蛋白及其mRNA的表达显著增高(P0.05);培养上清中MMP-9的浓度显著增高(P0.05),MMP-2的水平则无变化。不同的梯度组之间相比较,细胞迁移和侵袭能力、CD147的表达水平(蛋白及其mRNA)以及培养上清中MMP-9的浓度均随PA作用时间和作用剂量的增大而产生更显著的增高。结论:PA通过活化CD147-MMPs信号通路促进SMMC-7721细胞的迁移侵袭。     

Formation of Palmitic Acid/Ca2+ Complexes in the Mitochondrial Membrane: A Possible Role in the Cyclosporin-Insensitive Permeability Transition

A possible role of palmitic acid/Ca2+ (PA/Ca2+) complexes in the cyclosporin-insensitive permeability transition in mitochondria has been studied. It has been shown that in the presence of Ca2+, PA induces a swelling of mitochondria, which is not inhibited by cyclosporin A. The swelling is accompanied by a drop in membrane potential, which cannot be explained only by a work of the Ca2+ uniporter. With time, the potential is restored. Evidence has been obtained indicating that the specific content of mitochondrial lipids would favor the PA/Ca2+ -induced permeabilization of the membrane. In experiments with liposomes, the PA/Ca2+ -induced membrane permeabilization was larger for liposomes formed from the mitochondrial lipids, as compared to the azolectin liposomes. Additionally, it has been found that in mitochondria of the TNF (tumor necrosis factor)-sensitive cells (WEHI-164 line), the content of PA is larger than in mitochondria of the TNF-insensitive cells (C6 line), with this difference being mainly provided by PA incorporated in phosphatidylethanolamine and especially, cardiolipin. The PA/Ca2+ -dependent mechanism of permeability transition in mitochondria might be related to some pathologies, e.g. myocardial ischemia. The heaviness of myocardial infarction of ischemic patients has been demonstrated to correlate directly with the content of PA in the human blood serum.     

Palmitic Is the Main Fatty Acid Carried by Lipids of Detergent-Resistant Membrane Fractions from Neural and Non-Neural Cells

Lipids extracted from detergent-resistant membrane fractions, thought to derive from membrane domains, were analyzed for fatty acid composition. The proportion of palmitic acid in fractions isolated from neurons (cerebellar granule cells) and from neural-like cell lines (neuroblastoma-glioma NG108-15) nearly doubled (reaching about 54% of total fatty acids) with respect to cell WCL, indicating their enrichment in palmitic acid-carrying lipids. The proportion of palmitic acid in detergent-resistant fractions obtained from caveolin-transfected NG108-15 cells was comparable with that obtained from caveolin-negative cells, ruling out a specific role of this protein in recruiting palmitoylated lipid species. The enrichment in palmitic acid was remarked also in membrane fractions isolated from non-neuronal cell lines (A431) using either detergents or detergent-free techniques. Lipid fractionation and mass spectrometry experiments show that palmitic acid–rich phosphatidylcholine species are responsible of the peculiar fatty acid composition of these fractions. All together these results suggest that the enrichment in palmitic acid–rich phosphatidylcholine species is a common feature of neural and non-neural cell lines and may play a major role in the biogenesis of membrane domains.     

Enzymatic Synthesis of Cinnamic Acid Derivatives

Using Novozym 435 as catalyst, the syntheses of ethyl ferulate (EF) from ferulic acid (4-hydroxy 3-methoxy cinnamic acid) and ethanol, and octyl methoxycinnamate (OMC) from p-methoxycinnamic acid and 2-ethyl hexanol were successfully carried out in this study. A conversion of 87% was obtained within 2 days at 75 °C for the synthesis of EF. For the synthesis of OMC at 80 °C, 90% conversion can be obtained within 1 day. The use of solvent and high reaction temperature resulted in better conversion for the synthesis of cinnamic acid derivatives. Some cinnamic acid esters could also be obtained with higher conversion and shorter reaction times in comparison to other methods reported in the literature. The enzyme can be reused several times before significant activity loss was observed. Revisions requested 10 January 2006; Revisions received 17 January 2006     

The Enzymatic Production of Adipic Acid

Mutants of Brevibacterium sp. R312 were isolated for the production of adipic acid from 1,4-dicyanobutane (adiponitrile). One mutant (Ad), with a modified cell wall showed activity against adipamide three times greater than the wild type. Another mutant (ACV2) derived from the Ad strain had 30 times more activity on 5-cyanovaleric acid, and 7 times more on adipamide than the wild type.

The nitrile hydratase from the mutant strain ACV2 was purified and compared to that from the wild type R312. The nitrile hydratase of the mutant strain is different from that of the wild type by its pHi, optimum activity pH, and its rates of hydrolysis of 5-cyanovaleramide and 5-cyanovaleric acid which were 30 and 15 folds greater.

The presence of a new amidase named “adipamidase” acting on amide intermediates in the hydrolysis of dinitriles to organic acids was demonstrated in this mutant ACV2.