Bacterial catabolism of sulfanilic acid via catechol-4-sulfonic acid

Abstract A sulfanilic acid (4-aminobenzenesulfonic acid) degrading culture consisting of two strains (strain S1 and S2), was studied. Only strain S1 was able to attack sulfanilic acid. When strain S1 was cultavated in a mineral medium with sulfanilic acid an intensive violet colour was observed. The accumulating metabolite was isolated from the culture supernatant. By comparison with an authentic compound the metabolite was identified as catechol-4-sulfonic acid by thin layer and high performance liquid chromatography and by UV- and H-NMR spectroscopy. The occurrence of catechol-4-sulfonic acid indicates that there is no release of the sulfonic group before ring cleavage.     

Biodegradation of sulfanilic acid by Pseudomonas paucimobilis

An aerobic bacterium, isolated from a contaminated site, was able to degrade sulfanilic acid (4-aminobenzenesulfonic acid) and was identified as Pseudomonas paucimobilis. The isolate could grow on sulfanilic acid (SA) as its sole carbon and nitrogen source and metabolized the target compound to biomass. The bioconversion capacity depended on the sulfanilic acid concentration; greater than 98% elimination of the hazardous compound was achieved at low (10 mM) sulfanilic acid concentration, and the yield was greater than 70% at 50 mM concentration of the contaminant. The maximum conversion rate was 1.5 mmol sulfanilic acid/h per mg wet cells at 30 degrees C. Ca-alginate-phytagel proved a good matrix for immobilization of P. paucimobilis, with essentially unaltered biodegradation activity. Removal of sulfanilic acid from contaminated industrial waste water was demonstrated. SDS-PAGE analysis of the crude extract revealed novel proteins appearing upon induction with sulfanilic acid and related compounds, which indicated alternative degradation mechanisms involving various inducible enzymes.     

Degradation of azo dyes containing aminonaphthol by Sphingomonas sp strain 1CX

Sphingomonas sp strain 1CX was isolated from a wastewater treatment plant and is capable of aerobically degrading a suite of azo dyes, using them as a sole source of carbon and nitrogen. All azo dyes known to be decolorized by strain 1CX (Orange II, Acid Orange 8, Acid Orange 10, Acid Red 4, and Acid Red 88) have in their structure either 1-amino-2-naphthol or 2-amino-1-naphthol. In addition, an analysis of the structures of the dyes degraded suggests that there are certain positions and types of substituents on the azo dye which determine if degradation will occur. Growth and dye decolorization occurs only aerobically and does not occur under fermentative or denitrification conditions. The mechanism by which 1CX decolorizes azo dyes appears to be through reductive cleavage of the azo bond. In the case of Orange II, the initial degradation products were sulfanilic acid and 1-amino-2-naphthol. Sulfanilic acid, however, was not used by 1CX as a growth substrate. The addition of glucose or inorganic nitrogen inhibited growth and decoloration of azo dyes by 1CX. Attempts to grow the organism on chemically defined media containing several different amino acids and sugars as sources of nitrogen and carbon were not successful. Phylogenetic analysis of Sphingomonas sp strain 1CX shows it to be related to, but distinct from, other azo dye-decolorizing Sphingomonas spp strains isolated previously from the same wastewater treatment facility. Received 19 May 1999/ Accepted in revised form 11 August 1999     

AMPHOTERIC BEHAVIOR OF COMPLEX SYSTEMS : IV. NOTE ON THE ISOELECTRIC POINT AND IONIZATION CONSTANTS OF SULFANILIC ACID.

From the solubility minimum the value of the basic ionization constant of sulfanilic acid is shown to lie probably between the values 1.7 x 10^–15 and 3.2 x 10^–15. From solubility measurements the value of this same constant is shown to lie probably between 2.0 and 2.2 x 10^–15, and the isoelectric point of sulfanilic acid is thus at a cH of 0.056 or a pH of 1.25. From conductivity ratios the acid ionization constant of sulfanilic acid is shown to be 7.05 x 10^–4 at room temperature (21°C.). Calculations are made, from data published in preceding papers, of the ionization constants of glycine, K_a being 2.3 x 10^–10, and K_b being 2.2 x 10^–12.     

Sulfanilation of lysozyme by carbodiimide reaction. Reactivities of individual carboxyl groups

The carboxyl groups of lysozyme were coupled with sulfanilic acid, a chromophoric nucleophile, using 1-ethyl-3-dimethylaminopropylcarbodiimide at pH 5. Other carbodiimides were less effective. Ninety percent of the carboxyl groups were sulfanilated through exhaustive reaction with 1.2 m nucleophile. Isolation and identification of the tryptic peptides from this material showed that all 10 of the carboxyls of lysozyme had reacted. In 0.05 m sulfanilic, Glu-35 and Asp-101 were most reactive while Glu-7, Asp-18, and Asp-66 were least. Change to high concentration of nucleophile (from 0.05 to 1.2 m sulfanilic) altered carboxyl reactivity. Addition of inhibitor reduced reactivity of Asp-101 and Glu-35. Side reactions were not important.     

Isolation of a gene essential for biosynthesis of the lipopeptide antibiotics plipastatin B1 and surfactin in Bacillus subtilis YB8

Bacillus subtilis YB8 was found to produce the lipopeptide antibiotics surfactin and plipastatin B1. A gene, lpa-8, required for the production of both lipopeptides was cloned from strain YB8. When this gene was inactivated in strain YB8, neither surfactin nor plipastatin B1 was produced. However, the defective strain transformed with an intact lpa-8 gene had restored ability to produce both peptides. Nucleotide sequence analysis of the region essential for the production of the peptides revealed the presence of a large open reading frame. The deduced amino acid sequence of lpa-8 (224 amino acid residues) showed sequence similarity to that of sfp (from surfactin-producing B. subtilis), lpa-14 (from iturin A- and surfactin-producing B. subtilis), psf-1 (from surfactin-producing Bacillus pumilus), gsp (from gramicidin-S-producing Bacillus brevis), and entD (from siderophore-enterobactin-producing Escherichia coli), which are able to complement a defect in the sfp gene and promote production of the lipopeptide antibiotic surfactin. The sequence similarity among these proteins and the product similarity of cyclic peptides suggests that they might be involved in the biosynthesis or secretion of the peptides. Received: 14 July 1995 / Accepted: 22 December 1995     

A colorimetric assay for lignin based on its reaction with diazotized sulfanilic acid and its use in studies on lignin biodegradation by bacteria

Summary Studies on lignin biodegradation are considerably hampered by the lack of a simple analytical method. A rapid colorimetric assay for lignin has been developed using its reaction with a diazotized derivative of sulfanilic acid. Using such a method degradation of lignin, by an isolateErwinia sp. Cu3614, and by a genetically engineeredAcinetobacter PE7(pDPE2388) containing the arylether cleaving gene fromErwinia, has been followed.     

Leucine aminopeptidase as an ecto-enzyme of polymorphonuclear neutrophils

Intact polymorphonuclear neutrophils were modified chemically by a poorly permeable reagent, diazotized sulfanilic acid, and the changes in the activity of 5′-nucleotidase, alkaline phosphodiesterase, and leucine aminopeptidase were examined. Among three plasma membrane enzymes, 5′-nucleotidase activity was hardly detected in the human neutrophils. The activity of alkaline phosphodiesterase was observed in all the neutrophils examined, but was not inhibited by diazotized sulfanilic acid in the guinea-pig neutrophils. On the other hand, the activity of leucine aminopeptidase was not only found but also inhibited by diazotized sulfanilic acid without the inhibition of lactate dehydrogenase, a cytosol enzyme, in all the neutrophils, suggesting that leucine aminopeptidase is located generally on the plasma membrane as an ecto-enzyme in the neutrophils.     

Nouvelles données sur l'écologie et le comportement entomopathogène expérimental de l'entomophthoraleConidiobolus coronatus (Zygomycètes)

Résumé Quelle que soit l'origine de la souche (sol, arthropodes, lésions humaines), les conidies de l'EntomophthoraleConidiobolus coronatus sont capables d'infecter le puceronAcyrthosiphon pisum (Hémiptères, Aphididae) et les chenilles deGalleria mellonella (Lépidoptères, Pyralidae). De plus, les filtrats, de culture en milieu liquide agité des huit souches éprouvées sont toxiques par inoculation à l'égard de ces derniers insectes; cette toxicité se traduit d'abord par une hémorragie au trou de pip?re, qui peut durer plusiers heures, puis par un noircissement du corps de l'insecte et enfin par la mort de celui-ci. L'effet anticoagulant de substances produites par une Entomophthorale est ici signalé pour la première fois. Les deux souches les plus virulentes à l'égard deAcyrthosiphon pisum, parmi lesquelles se trouve celle isolée de lésions humaines, sont également les plus toxinogènes. L'étude de la vitesse de croissancein vitro du champignon à des températures comprises entre 4 et 37°C révèle une adaptation des souches aux conditions thermiques en relation avec la région d'origine de l'isolat. Notamment, les souches d'origine tropicale se développent plus, rapidement à 26–29°C que les souches originaires de France. La vitesse maximale de croissance est constatée à 29°C pour les différentes souches, sauf pour celle isolée de lésions humaines qui se développe le plus rapidement à 37°C. Cette température est létale pour les autres souches sauf pour une souche isolée d'un myriapode de l'?le de la Réunion. C'est la première fois que l'aptitude à cro?tre à cette température est reconnue chez un isolat deConidiobolus coronatus provenant d'un arthropode.

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Summary Whatever the substrate of origin of the strain (soil, arthropods, human phycomycosis), conidia ofConidiobolus coronatus (Zygomycotina, Entomophthorales) are able to infect the pea aphid,Acyrthosiphon pisum (Hemiptera, Aphididae) and the greater wax moth,Galleria mellonella (Lepidoptera, Pyralidae). Moreover, intrahemocoelic injections of filtrates from shake cultures of all the 8 strains tested proved to be toxic for the later insect; the symptoms are successively haemorrhage, which could last for a few hours, blackening of the blood and at last death. This is the first mention of an anti-coagulant effect of metabolic products from any Entomophthorale. The strain isolated from an human phycomycosis is both the most virulent against the pea aphid and the most effective with regard to toxicity. Comparison of fungal growthin vitro at different temperatures, ranging from 4 to 37°C, revealed a certain relation between the temperature for the optimal growth and the locality of origin of a strain. In particular, strains from tropical areas were growing at 26–29°C faster than those from temperate regions. The higher growth rate was noticed at 29°C for all the 8 tested strains, except for the strain from human phycomycosis, for which the optimum temperature was 37°C. This temperature was lethal for the other strains, but for a strain isolated from a myriapod from the Reunion Island. It is the first time that the ability of a strain ofC. coronatus isolated from an arthropod to growth at 37°C is ascertained.

     

Leucine aminopeptidase as an echo-enzyme of polymorphonuclear neutrophils

Intact polymorphonuclear neutrophils were modified chemically by a poorly permeable reagent, diazotized sulfanilic acid, and the changes in the activity of 5'-nucleotidase, alkaline phosphodiesterase, and leucine aminopeptidase were examined. Among three plasma membrane enzymes, 5'-nucleotidase activity was hardly detected in the human neutrophils. The activity of alkaline phosphodiesterase was observed in all the neutrophils examined, but was not inhibited by diazotized sulfanilic acid in the guinea-pig neutrophils. On the other hand, the activity of leucine aminopeptidase was not only found but also inhibited by diazotized sulfanilic acid without the inhibition of lactate dehydrogenase, a cytosol enzyme, in all the neutrophils, suggesting that leucine aminopeptidase is located generally on the plasma membrane as an ecto-enzyme in the neutrophils.     

Biological activity of secondary metabolites produced by a strain of<Emphasis Type="Italic">Pseudomonas fluorescens</Emphasis>

Biological activity of secondary metabolites produced by a plant-growth-promotingPseudomonas fluorescens was evaluated. The strain produced antibiotics phenazine (PHE), 2,4-diacetylphloroglucinol (PHL) and siderophore pyoverdin (PYO) in standard King’s B and succinic acid media, respectively. After extraction, PYO was identified by comparing the UV-spectra and moss-green color development after ‘diazotized sulfanilic acid’ (DSA) spray in TLC. PHE and PHL were identified by comparing standard compounds on TLC and orange-color development immediately after DSA spray.In vitro antibiosis study of the metabolites revealed their antibacterial and antifungal activity against bacterial test organismsCorynebacterium sp.,Mycobacterium phlei andM. smegmatis and test fungiFusarium moniliforme, F. oxysporum, F. semitectum, F. solani andRhizoctonia solani. A statistically significantly higher plant growth was recorded in siderophore-amended plantlets under gnotobiotic conditions whereas PHE and PHL did not show any plant-growth-promoting activity. These results support the importance of the secondary metabolites produced by the strainP. fluorescens in enhancing plant growth and in controling fungal and bacterial pathogens.     

Properties of sulfanilazo-haptoglobin

1. Tyrosine and two structural isomers of histidine residues in human haptoglobin were modified with diazotized sulfanilic acid. Sulfanilazo-derivatives of haptoglobin obtained by increasing the reagent/protein molar ration showed gradual decrease of peroxidase activity when complexed with hemoglobin. 2. Formation of haptoglobin derivatives with ten mono(sulfanilazo)-tyrosines and two mono (sulfanilazo)histidines resulted in the blockage of one out of six antigenic determinants, whereas immunoreactivity of the derivative with fourteen azotyrosines, one C-4, and two C-2 azohistidines was decreased by half. 3. Removal of sialic acid from oligosaccharide chains of haptoglobin made the molecule more accessible to diazotized sulfanilic acid. 4. Sulfanilazo-modification of tyrosine and histidine residues was practically of no effect in the reaction of haptoglobin with plant lectin, concanavalin A.     

Characterization of a New <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> Strain NJ-15 as a Potential Biocontrol Agent

Phylogenetic characterization of soil isolate NJ-15, based on sequence homology of a partial 746-bp fragment of 16SrDNA amplicon, with the ribosomal database sequences (http://www.msu.edu/RDP/cgis/phylip.cgi), validated the strain as Pseudomonas aeruginosa. The strain NJ-15 produced a substantial amount of indole acetic acid (IAA) in tryptophan-supplemented medium. Besides, the strain also exhibited significant production of both the siderophore and hydrogen cyanide (HCN) on chrome azurol S and King's B media, respectively. The data revealed lower HCN production under iron-limiting conditions vis-à-vis higher HCN release with iron stimulation. Significant growth inhibition of phytopathogenic fungi occurred in the order as Fusarium oxysporum > Trichoderma herizum > Alternaria alternata > Macrophomina phasiolina upon incubation with strain NJ-15 cells. Thus, the secondary metabolites producing new Pseudomonas aeruginosa strain NJ-15 exhibited innate potential of plant growth promotion and biocontrol activities in vitro. Received: 30 April 2002 / Accepted: 5 July 2002     

Chemical treatment of macrophages increases their responsiveness to migration inhibitory factor (MIF).

The response of guinea pig macrophages to migration inhibitory factor (MIF) is altered by several chemical treatments. Treatment of macrophages with the diazonium salt of sulfanilic acid (5 x 10(-6) to 4 x 10(-4) M) significantly increases the response of these cells to MIF. Treatment with acetic anhydride also augments the response of these cells to MIF. The latter finding suggests that alteration of amino, hydroxyl, or sulfhydryl groups is involved in this phenomenon. Treatment of macrophages with sodium periodate (2 x 10(-5) to 10(-3) M) which is known to oxidize cis-glycols and with hydroxylamine (2 x 10(-5) to 2 x 10(-3) M), which reacts with carbonyl groups also increases response to MIF. The following experiments suggest that the significant alteration occurs at the level of the cell surface. Incubation of macrophages with the diazonium salt of sulfanilic acid at 4 degrees C, at which temperature pinocytosis is largely inhibited, is sufficient to increase the MIF response. The activity of the cytoplasmic enzyme aspartate aminotransferase, which in homogenates is susceptible to inactivation by low concentrations of the diazonium salt of sulfanilic acid, is not decreased when intact macrophages are incubated with high concentrations of the diazonium salt of sulfanilic acid. Cumulatively, these findings suggest that modification of different functional groups on the macrophage surface causes the same physiologic effect.     

Studies on the Hypertrophic Disease of Cherry (Genus Prunus), So-called “Witch’s Broom” Caused by Taphrina wiesneri

Metabolites of Taphrina wiesneri (Rath.) Mix. were examined. Brassicasterol, stearic acid, and p-hydroxyphenylacetic acid were isolated in crystalline form. p-Hydroxybenzoic acid and vanillic acid were identified by paper chromatography and UV measurement. Palmitic acid was identified by gas-chromatography. The fungus produced usually these compounds on any one of four kinds of medium used. p-Hydroxyphenylacetic acid promoted germination of rape seeds at the concentration of 20 ppm in water and showed inhibition at 250 ppm.

Phenolic acids and their related compounds in Japanese flowering cherry leaves infected by Taphrina wiesneri were examined. In the acidic and neutral extracts of infected cherry leaves (I), eighteen compounds positive to diazotized sulfanilic acid and two fluorescent compounds were detected by paper chromatography. Of these compounds, coumarin, 3, 4-dihydrocoumarin, melilotic acid, o- and p-coumaric acids, p-hydroxybenzoic melilotic acid, ferulic acid and caffeic acid were identified. Melilotic acid and coumarin were obtained in crystalline form. The amount of melilotic acid in I was higher than that in healthy leaves independent of sample source, although increased with the growth of cherry leaves.     

Screening of the insecticidal activity ofBacillus thuringiensis strains against the egyptian cotton leaf wormSpodoptera littoralis

A screening of the larvicidal activity of the more than 900 strains ofBacillus thuringiensis strains, combining the Institut Pasteur collection was realized. A quick bioassay using 1st instar larvae and semi-synthetic medium was developed. Many strains were toxic toSpodoptera littoralis, but only a few belonging mainly to serovarsaizawai, kenyae andentomocidus showed high level of toxicity. The profiles of strain activities differed from serovar to serovar, but within the same serovar toxicity can vary with different strains. Oneaizawai strain tested in the field gave satisfactory results, better than a commercially used strain, tested in the same experiment.

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Résumé Le criblage de notre collection deB. thuringiensis (plus de 900 souches) a été effectué contreSpodoptera littoralis. Une technique rapide de bioessai utilisant des chenilles néonates et un milieu semisynthétique a été mise au point. Beaucoup de souches se sont montrées actives à forte dose, mais seulement quelques-unes appartenant principalement aux sérovaraizawai etkenyae à doses plus faibles. Les différents sérotypes ont un profil d'activité différent mais un même sérotype peut comprendre des souches de toxicité variée. Une souche du sérovaraizawai a été testée en champs et a donné de bons résultats, son activité ayant été évaluée à environ 5 fois celle d'une souche couramment commercialisée.

     

Specificity and affinity of neuraminic acid exhibited by canine rotavirus strain K9 carbohydrate‐binding domain (VP8*)

The outer capsid spike protein VP4 of rotaviruses is a major determinant of infectivity and serotype specificity. Proteolytic cleavage of VP4 into 2 domains, VP8* and VP5*, enhances rotaviral infectivity. Interactions between the VP4 carbohydrate‐binding domain (VP8*) and cell surface glycoconjugates facilitate initial virus‐cell attachment and subsequent cell entry. Our saturation transfer difference nuclear magnetic resonance (STD NMR) and isothermal titration calorimetry (ITC) studies demonstrated that VP8*_64‐224 of canine rotavirus strain K9 interacts with N‐acetylneuraminic and N‐glycolylneuraminic acid derivatives, exhibiting comparable binding epitopes to VP8* from other neuraminidase‐sensitive animal rotaviruses from pigs (CRW‐8), cattle (bovine Nebraska calf diarrhoea virus, NCDV), and Rhesus monkeys (Simian rhesus rotavirus, RRV). Importantly, evidence was obtained for a preference by K9 rotavirus for the N‐glycolyl‐ over the N‐acetylneuraminic acid derivative. This indicates that a VP4 serotype 5A rotavirus (such as K9) can exhibit a neuraminic acid receptor preference that differs from that of a serotype 5B rotavirus (such as RRV) and the receptor preference of rotaviruses can vary within a particular VP4 genotype.     

AMPHOTERIC BEHAVIOR OF COMPLEX SYSTEMS : III. THE CONDUCTIVITY OF SULFANILIC ACID-LYSIN MIXTURES.

Conductivities of sulfanilic acid, lysin, and mixtures of the two were made over a wide pH range, the pH being adjusted by means of phosphate buffers. The actual conductivities of the sulfanilic acid, the lysin, and the mixture were calculated. The difference between the conductivity of the mixture and the sum of the conductivities of the components alone passes through a maximum at a pH theoretically calculable as the isoelectric point of the system. Certain applications of the results are made to the explanation of the behavior of living tissues.     

Induction and Citric Acid Productivity of Fluoroacetate-sensitive Mutant Strains of Candida lipolytica

To establish a novel process for the economical production of citric acid from n-paraffins by yeast, attempts were made to obtain some mutant strains capable of producing citric acid in higher yield without (+)-isocitric acid.

From among the mutant strains derived from Candida lipolytica ATCC 20114, which produced citric acid and (+)-isocitric acid in the ratio of about 60:40 from n-paraffins, a citrate non-utilizing mutant strain, K-20, and a fluoroacetate-sensitive mutant strain , S-22, were selected on the basis of high citric acid and low (+)-isocitric acid productivity.

The mutant strain S-22 showed extremely poor growth in a medium containing sodium citrate as the sole carbon source and extremely high sensitivity to fluoroacetate. The production ratio of citric acid and (+)-isocitric acid by the mutant strain was changed to 97:3, and the yield of the citric acid from n-paraffins, charged to the fermentation medium, reached 145%(w/w).