Effects of 3,4-dihydroxybenzoic acid on root membrane potential in micropropagated tobacco plants

In order to understand the mechanism of action of the phenolic compound 3,4-dihydroxybenzoic acid, we tested its effect on tobacco root membrane potential. Tobacco root segments, excised from micropropagated plants grown in liquid media, were perfused with 0.1–5 mM 3,4-dihydroxybenzoic acid. Activity on the plasma membrane potential was compared with that obtained after perfusion with 0.05 mM indole-3-acetic acid, 0.05 mM kinetin and 0.05 mM gibberellic acid. Possible interactions between 3,4-dihydroxybenzoic acid and plant growth regulators were evaluated by the means of successive applications. When applied to tobacco root segments, 3,4-dihydroxybenzoic acid elicited a transient membrane depolarization. The membrane depolarization induced by 3,4-dihydroxybenzoic acid was followed by a repolarization phase, as for auxin applications. In roots preconditioned with the other growth regulators, the activity of 3,4-dihydroxybenzoic acid on membrane potential was non-specifically affected. In roots preconditioned with 3,4-dihydroxybenzoic acid, indole-3-acetic acid activity on cell membrane was altered, suggesting a specific reciprocal interaction. This revised version was published online in June 2006 with corrections to the Cover Date.     

Bacterial degradation of 3,4,5-trimethoxyphenylacetic and 3-ketoglutaric acids.

When grown at the expense of 3,4,5-trimethoxyphenylacetic acid, a species of Arthrobacter readily oxidized 3,4-dihydroxy-5-methoxyphenylacetic acid, but other structurally related aromatic acids were oxidized only slowly. Cell extracts contained a dioxygenase for 3,4-dihydroxy-5-methoxyphenylacetate, and the corresponding trihydroxy acid, which was not attacked by the enzyme, inhibited oxidation of this ring-fission substrate. Cell suspensions did not release carbon dioxide from 3,4-[methoxyl-14C]dihydroxy-5-methoxyphenylacetate but accumulated 1 mol of methanol per mol of 3,4,5-trimethoxyphenylacetate oxidized. A cell extract converted the ring-fission substrate into stoichiometric amounts of pyruvate and acetoacetate, formed from 3-ketoglutarate by the action of an induced decarboxylase. 3-Ketoglutaric acid served as sole source of carbon for many soil isolates.     

红背山麻杆叶的化学成分研究(Ⅰ)——酚酸类及相关化合物

采用80%丙酮提取石油醚萃取部位,利用凝胶、MCI及Toyopearl Butyl-650C柱色谱进行分离纯化得到10个酚酸类及相关化合物。根据化合物的波谱数据分析鉴定为水杨酸(1)、对羟基苯甲酸(2)、2,5-二羟基苯甲酸(3)、3,4-二羟基苯甲酸(4)、反-对香豆酸(5)、顺-对香豆酸(6)、咖啡酸(7)、咖啡酸甲酯(8)、没食子酸(9)、没食子酸甲酯(10)。其中化合物1~8、10均为首次从本属植物中分离得到。     

The phenolic, 3,4-dihydroxybenzoic acid,is an endogenous regulator of rooting in <Emphasis Type="Italic">Protea cynaroides</Emphasis>

Analysis of stem extracts identified large quantities of 3,4-dihydroxybenzoic acid and other similar phenolics. The exogenous application of 3,4-dihydroxybenzoic acid on Protea cynaroides explants in vitro significantly increased the root mass at 100 mg l⁻¹, but not at lower concentrations, while root inhibition was observed at 500 mg l⁻¹. HPLC analysis of cuttings during vegetative propagation showed a considerable increase in 3,4-dihydroxybenzoic acid levels from initial planting to when root formation took place, indicating for the first time that 3,4-dihydroxybenzoic acid may be an important phenolic compound in regulating root formation in P. cynaroides cuttings. HPLC analysis also identified caffeic, ferulic, gallic and salicylic acids in the cuttings.     

Degradation of I-Naphthol by a Soil Pseudomonad

A soil Pseudomonas sp. grew with 1-naphthol as sole organic carbon source and produced a 3,4-dihydro-dihydroxy-1(2H)-naphthalenone as the main early intermediary metabolite. Washed 1-naphthol-grown organisms oxidized naphthalene, 1- or 2-naphthol, salicylic acid and, to some extent, 2,3-dihydroxybenzoic acid.     

Pigment Production of Cryptococcus neoformans Grown with Extracts of Guizotia abyssinica

Brown pigment(s) formed in Cryptococcus neoformans when grown on media containing extracts of the seeds of Guizotia abyssinica cannot be extracted by common organic solvents or by 6 n HCl or 2 n NaOH. A similar pigmentation was observed in C. neoformans when grown on a medium containing caffeic acid isolated from the hydrolyzed methanol extract of G. abyssinica seeds. Its methyl ester and the diacetate thereof, as well as the following structurally related compounds, 3-hydroxytyramine, 3,4-dihydroxybenzoic acid, 3,4-dihydroxyphenylethanolamine, and 4-hydroxy-3,5-dimethoxycinnamic acid, brought about similar pigmentation. However, 2,4-, 2,5-, 2,6-, and 3,5-dihydroxybenzoic acids, tyrosine, phenylalanine, cinnamic acid, 4-hydroxycinnamic acid, and 4-hydroxy-3-methoxycinnamic acid did not cause coloration in C. neoformans.     

Excretion of anthranilate and 3-hydroxyanthranilate by Saccharomyces cerevisiae: relationship to iron metabolism.

Resting suspensions of cells of Saccharomyces cerevisiae grown in iron-rich or iron-deficient conditions were studied by following the fluorescence emission changes (lambda em. 400-460 nm, lambda exc. 300-340 nm) occurring in these suspensions upon addition of glucose and ferric iron. The results show that, in addition to NAD(P)H, metabolites of the aromatic amino acid pathway interfere with the fluorescence measurements, and that they could be involved in ferric iron reduction. Wild-type strains of S. cerevisiae are known to excreted anthranilic acid and 3-hydroxyanthranilic acid in response to glucose. The major fluorescing compound excreted by a chorismate-mutase-deficient mutant strain of S. cerevisiae was identified as anthranilic acid. The excretion of anthranilic and 3-hydroxyanthranilic acids was correlated with the ferric-reducing capacity of the extracellular medium. Excretion during growth was much greater by cells cultured in iron-rich medium than by cells grown in iron-deficient medium. The possibility was examined that a link could exist between the biosynthesis of aromatics and the ferri-reductase activity of the cells, via chorismate synthase and its putative diaphorase-associated activity. Two ferri-reductase-deficient mutants excreted much less 3-hydroxyanthranilate than did the parental wild-type strains. However, the ferri-reductase activity of a chorismate-synthase-deficient mutant was comparable to that of the parental strain.     

Studies in vitro on the involvement of O-sulphate esters in the formation of O-methylated 3,4-dihydroxybenzoic acid by rat liver.

The involvement of O-sulphate esters in the directed O-methylation was investigated in vitro with a dialysed "high-speed' supernatant from rat liver as the enzyme preparation and the catechol compound 3,4-dihydroxybenzoic acid as the substrate. The enzyme reactions involved were studied separately with the O-methylated and O-sulphated derivatives. The rate of hydrolysis by arylsulphatase was 14.5 nmol/min per mg of protein for 3-methoxy-4-sulphonyloxybenzoic acid and 10.1 nmol/min per mg of protein for 4-methoxy-3-sulphonyloxybenzoic acid. The sulphotransferase activity towards the guaiacols 4-hydroxy-3-methoxybenzoic acid and 3-hydroxy-4-methoxybenzoic acid was 570pmol of 4-O-sulphated and 350pmol of 3-O-sulphated product formed/min per mg of protein. The 3-O- and 4-O-sulphate esters of 3,4-dihydroxybenzoic acid could not serve as substrates for the catechol O-methyltransferase reaction. When either ester was incubated in the presence of S-adenosyl-L-methionine, but without the arylsulphatase inhibitor KH2PO4, 3,4-dihydroxybenzoic acid was formed, which was subsequently O-methylated in a meta/para ratio of 4.6. It is concluded that O-methylation can precede O-sulphation but that O-sulphation prevents further metabolism by O-methylation. Also O-sulphate esters do not have a directing effect on O-methylation. From the study of the simultaneous action of sulphotransferase and catechol O-methyltransferase on 3,4-dihydroxybenzoic acid we conclude that O-sulphation and O-methylation proceed independently of each other under the assay conditions used, both directed preferentially to the 3-hydroxy group.     

一株白粉寄生菌的酚类代谢产物

从得自鼎湖山自然保护区的一株白粉寄生菌(Ampelomyces sp.)SC0307固体发酵物中分离得到7个酚类化合物.通过波谱分析,分别鉴定为2,5-二羟基苯甲醇(1)、对羟基苯甲酸(2)、2,5-二羟基苯甲酸(3)、3,4-二羟基苯甲酸(4)、苯乙酸(5)、3,4-二甲氧基肉桂酸(6)、3,4,5-三甲氧基肉佳酸(7).7个化合物均为从白粉寄生菌属真菌中首次分离获得.     

Itoic Acid Synthesis in Bacillus subtilis

Under conditions of iron deficiency, strains of Bacillus subtilis produced 2,3-dihydroxybenzoic acid (DHB), 2,3-dihydroxybenzolyglycine (DHBG), or both of these compounds. DHB(G) production [production of DHB(G) refers to the production of DHB, or DHBG, or both] was proportional to the amount of iron present and occurred logarithmically, paralleling growth. Supplementation of media with more than 150 mug of iron per liter at zero-time inhibited DHB accumulation completely. In the presence of DHB, lower levels of iron inhibited DHB(G) production, so that the actual inhibitor of synthesis may involve the Fe(3+):[DHB(G)](3) complex. The strains producing DHBG also produced coproporphyrin III during iron-deficient growth, whereas a strain producing DHB did not produce coproporphyrin III under these conditions. Accumulation of DHB(G) was influenced by the levels of aromatic amino acids and anthranilic acid in the medium. In vivo experiments with strain B-1471 demonstrated that DHB was coupled to added glycine to form DHBG. Metabolism of DHB(G) was observed in two of the strains studied.     

Anaerobic degradation of 2-aminobenzoic acid (anthranilic acid) via benzoyl-coenzyme A (CoA) and cyclohex-1-enecarboxyl-CoA in a denitrifying bacterium.

The enzymes catalyzing the initial reactions in the anaerobic degradation of 2-aminobenzoic acid (anthranilic acid) were studied with a denitrifying Pseudomonas sp. anaerobically grown with 2-aminobenzoate and nitrate as the sole carbon and energy sources. Cells grown on 2-aminobenzoate are simultaneously adapted to growth with benzoate, whereas cells grown on benzoate degrade 2-aminobenzoate several times less efficiently than benzoate. Evidence for a new reductive pathway of aromatic metabolism and for four enzymes catalyzing the initial steps is presented. The organism contains 2-aminobenzoate-coenzyme A ligase (2-aminobenzoate-CoA ligase), which forms 2-aminobenzoyl-CoA. 2-Aminobenzoyl-CoA is then reductively deaminated to benzoyl-CoA by an oxygen-sensitive enzyme, 2-aminobenzoyl-CoA reductase (deaminating), which requires a low potential reductant [Ti(III)]. The specific activity is 15 nmol of 2-aminobenzoyl-CoA reduced min-1 mg-1 of protein at an optimal pH of 7. The two enzymes are induced by the substrate under anaerobic conditions only. Benzoyl-CoA is further converted in vitro by reduction with Ti(III) to six products; the same products are formed when benzoyl-CoA or 2-aminobenzoyl-CoA is incubated under reducing conditions. Two of them were identified preliminarily. One product is cyclohex-1-enecarboxyl-CoA, the other is trans-2-hydroxycyclohexane-carboxyl-CoA. The complex transformation of benzoyl-CoA is ascribed to at least two enzymes, benzoyl-CoA reductase (aromatic ring reducing) and cyclohex-1-enecarboxyl-CoA hydratase. The reduction of benzoyl-CoA to alicyclic compounds is catalyzed by extracts from cells grown anaerobically on either 2-aminobenzoate or benzoate at almost the same rate (10 to 15 nmol min-1 mg-1 of protein). In contrast, extracts from cells grown anaerobically on acetate or grown aerobically on benzoate or 2-aminobenzoate are inactive. This suggests a sequential induction of the enzymes.     

The identification and biosynthesis of siderochromes formed by Micrococcus denitrificans.

1. Micrococcus denitrificans excretes three catechol-containing compounds, which can bind iron, when grown aerobically and anaerobically in media deficient in iron, and anaerobically in medium with a high concentration of Ca2+. 2. One of these compounds was identified as 2,3-dihydroxybenzoic acid (compound I), and the other two were tentatively identified as N1N8-bis-(2,3-dihydroxybenzoyl)spermidine (compound II) and 2-hydroxybenzoyl-N-L-threonyl-N4[N1N8-bis-(2,3-dihydroxybenzoyl)]spermidine (compound III). 3. The equimolar ferric complex of compound III was prepared; compound III also forms complexes with Al3+, Cr3+ and Co2+ ions. 4. Cell-free extracts from iron-deficient organisms catalyse the formation of compound II from 2,3-dihydroxybenzoic acid and spermidine, and of compound III from compound II, L-threonine and 2-hydroxybenzoic acid; both reactions require ATP and dithiothreitol, and Mg2+ stimulates activity. The enzyme system catalysing the formation of compound II has optimum activity at pH 8.8 Fe2+ (35muM), Fe3+ (35muM) and Al3+ (65muM) inhibit the reaction by 50 percent. The enzyme system forming compound III has optimum activity at pH 8.6. Fe2+ (110 muM), Fe3+ (110 muM) and Al3+ (135 muM) inhibit the reaction by 50 percent. 5. At least two proteins are required for the formation of compound II, and another two proteins for its conversion into compound III. 6. The changes in the activities of these two systems were followed after cultures became deficient in iron. 7. Ferrous 1,10-phenanthroline is formed when a cell-free extract from iron-deficient cells is incubated with the ferric complex of compound III, succinate, NADH and 1,10-phenanthroline under N2.     

Influence of metal ions on the formation of mycobactin and salicylic acid in Mycobacterium smegmatis grown in static culture

Mycobacterium smegmatis was grown on trace-metal-free medium in static culture. Throughout the growth phase, the concentration of mycobactin increased continuously, reaching a maximum of about 30 to 40 mug of mycobactin/mg of cell dry weight after 6 days; the concentration of salicylic acid remained approximately constant at 1 to 2 mug of salicylic acid/mug of cell dry weight. Fe(2+) (or Fe(3+)), Zn(2+), Mn(2+), and Mg(2+) were all essential to a maximum formation of mycobactin. Optimum concentrations required were: Fe(2+), about 1.8 mum; Mn(2+) and Zn(2+), about 0.5 mum; and Mg(2+), at least 0.17 mm. Higher levels of Fe(2+) (9 to 90 mum) and Zn(2+) (2 to 7 mum) repressed mycobactin to about half the maximum value. No other cation or anion apparently is required for mycobactin biosynthesis. Salicylic acid concentration increased about fourfold when iron was omitted from the medium, but this is not as great as the increase reported previously for this strain of M. smegmatis. Mycobactin formation in another strain of M. smegmatis, NCIB 8548, showed similar dependencies on Fe(2+), Zn(2+), and Mn(2+). Maximum accumulation of mycobactin with this strain was 85 mug of mycobactin/mg of dry cell weight, under iron-deficient (1.8 mum Fe(2+)) conditions.     

Effect of phenolic acids on anthocyanin content in maize roots

Supply of 0.01 to 5.0 mM salicylic, caffeic and gallic acids, either during imbibition of seeds for 24 to 48 h or during seedling growth increased anthocyanin production in maize (Zea mays L. cv. Ganga safed-2) roots. While tyrosine had no effect, phenylalanine either in the presence or absence of the phenolic acids increased anthocyanin content. Glucose in a concentration range of 1 to 20 mM and shikimic acid in 0.01 to 5.0 mM range also increased pigment level, which was higher in the presence of salicylic acid than in it.s absence. The experiments demonstrate the possibility of some indirect effects of salicylic acid and other phenolic acids on anthocyanin synthesis.     

几种酶活抑制剂对红曲霉色素合成的影响

红曲色素是天然安全的色素和防腐剂,根据代谢数据库选择了6种代谢途径关键酶的抑制剂,在基本培养基中考察这些抑制剂对红曲霉生长和合成色素的影响。甲羟戊酸合成途径的抑制剂邻氨基苯甲酸和3,4-二羟苯甲酸对红曲霉生长和色素生物合成都没有影响;莽草酸途径关键酶氨基苯甲酸合成酶的抑制剂三甲胺不抑制红曲霉的生长和色素的合成。在不影响红曲霉生长的浓度范围内,聚酮途径中β-酮酯酰-ACP合成酶的专性抑制剂碘乙酰胺(0.5mmol/L)抑制红曲色素合成程度达64.7%,非专性抑制剂咪唑(1mmol/L)抑制幅度达60%,聚酮途径硫酯酶的抑制剂2,4-二硝基氟苯(0.5mmol/L)强烈抑制红曲霉合成色素的活性,抑制程度达91.5%。相关酶活抑制的试验数据显示红曲霉可能经过聚酮途径合成红曲色素。     

Pigment Formation for Differentiating Cryptococcus neoformans from Candida albicans

When 2,3- or 3,4-dihydroxybenzoic acid, 3,4-dihydroxyphenylalanine, and 3,4-dihydroxycinnamic acid are added to growth media, they are converted to a characteristic brown pigment by Cryptococcus neoformans. This pigment formation has hitherto been encountered only when this microorganism was cultivated on media containing Guizotia abyssinica seed. This phenomenon can be used for differentiating Cryptococcus neoformans from Candida albicans. Possible precursors of these o-diphenols (quinic acid, aromatic monohydroxy acids, or tyrosine) do not give rise to the brown pigmentation.     

Biotransformation of 3-methylphthalate by Micrococcus sp. strain 12B

When Micrococcus strain 12B grown on o-phthalate was incubated with 3-methylphthalate, three compounds accumulated. These were shown to be 2-pyrone-3-methyl-4,6-dicarboxylic acid, 3,4-dihydroxy-6-methylphthalic acid, and 5-hydroxy-3-methyphthalic acid, all previously undescribed. A pathway for the formation of these compounds is proposed.     

Iridescent material and the effect of iron on its production byPseudomonas aeruginosa

The nutritional conditions controlling iridescence inPseudomonas aeruginosa were studied using synthetic media solidified with agar. Iron and magnesium were growth-limiting factors in media solidified with dialysed agar. Iridescence only occurred on iron-deficient media and was not suppressed by adding Ca, Cu, Mn and Zn to these media. The ultraviolet absorption spectrum of the iridescent material was almost identical to the spectrum of the pyo I substances which are 2-alkyl-4-quinolinols.The amount of material produced was inversely proportional to the iron content of the medium. Small amounts of material were produced by cells grown at levels of iron optimal for growth. Synthesis of 2-alkyl-4-quinolinol may be a normal metabolic process in the iridescent strains ofPseudomonas aeruginosa. It was enhanced by anthranilic acid and tryptophan; kynurenine and kynurenic acid had no effect. The results can be explained if it is assumed that the activity of iron-requiring enzymes catalizing the breakdown of tryptophan is reduced.Even in the presence of anthranilic acid or tryptophan no material was produced by a non-iridescent strain.     

The isolation and identification of 3,4-dihydroxybenzoic acid formed by nitrogen-fixing Azomonas macrocytogenes

Abstract The mechanism of iron acquisition was studied in nitrogen-fixing Azomonas macrocytogenes . Spent solid agar plating medium samples from Fe-deficient and Fe-sufficient cultures were subjected to the high voltage paper electrophoresis siderophore assay. A yellow-green fluorescent peptide was elicited only under conditions of iron deficiency, whereas a novel dark blue fluorescent, Arnowpositive phenolic compound was detected under both Fe-deficient and Fe-sufficient conditions. Both compounds formed colored complexes with ferric iron. SDS-PAGE analysis of outer envelope protein preparations revealed the hyperproduction of an 83 kDa protein under iron-limiting conditions. Chemical analysis indicated that the phenolate was 3,4-dihydroxybenzoic acid (protocatechuic acid), a compound previously unreported as an extracellular product of a diazotroph.     

Inhibition of Aspergillus growth and aflatoxin release by derivatives of benzoic acid.

A study was conducted to determine the effects of o-nitrobenzoate, p-aminobenzoate, benzocaine (ethyl aminobenzoate), ethyl benzoate, methyl benzoate, salicylic acid (o-hydroxybenzoate), trans-cinnamic acid (beta-phenylacrylic acid), trans-cinnamaldehyde (3-phenylpropenal), ferulic acid (p-hydroxy-3-methoxycinnamic acid), aspirin (o-acetoxy benzoic acid), and anthranilic acid (o-aminobenzoic acid) upon growth and aflatoxin release in Aspergillus flavus NRRL 3145 and A. parasiticus NRRL 3240. A chemically defined medium was supplemented with various concentrations of these compounds and inoculated with spores, and the developing cultures were incubated for 4, 6, and 8 days at 27 degree C in a mechanical shaker. At the beginning of day 8 of incubation, aflatoxins were extracted from cell-free filtrates, separated by thin-layer chromatography, and quantitated by ultraviolet spectrophotometry. The structure of these aromatic compounds appeared to be critically related to their effects on mycelial growth and aflatoxin release. At concentrations of 2.5 and 5.0 mg per 25 ml of medium, methyl benzoate and ethyl benzoate were the most effective in reducing both mycelial growth and aflatoxin release by A. flavus and A. parasiticus. Inhibition of mycelial growth and aflatoxin release by various concentrations of the above-named aromatic compounds may indicate the possibility of their use as fungicides.