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PubMed1983

[Effect of prodigiosin and its combination with immunodepressants on the graft versus host reaction in mice].

The local (lymph node) graft-versus-host reaction (GVHR) in F1 (CBA X C57BL/6) mice and the lethal GVHR in C57BL/6 mice were induced by transfer of lymph node cells of CBA mice with skin allotransplants from C57BL/6 mice. Prednisolone in combination with asathioprin (imuran) administered to CBA mice inhibited the GVHR. Prodigiosan used alone was not active, while in combination with immunodepressants it increased their inhibitory effect. Adhesive cells with a suppressive activity were detected in the spleen of mice treated with prodigiosan. Such cells were capable of suppressing the capacity of syngeneic lymphocytes for inducing the GVHR.

Animals↗
PubMed1982

Prodigiosin, a component of kappa phage receptor complex in Serratia marcescens.

Studies on kappa phage inactivation with isolated pigments from Serratia marcescens were carried out. Kappa phage is sensitive to inactivation with diethyl ether, petroleum ether, acetone and methanol, but is quite stable in chloroform and dimethyl sulphoxide. The pigment extract dissolved in chloroform inactivates about 50% of the total suspended phages. The pigment dissolved in acetone and dimethyl sulphoxide inactivates about 96.50% and 64.10% of the phages, respectively. High inactivation values with acetone were partially due to direct inactivation rather than the pigment itself.

Bacteriophages↗
PubMed1971

Induction of pigmentation in nonproliferating cells of Serratia marcescens by addition of single amino acids.

Addition of casein hydrolysate to suspensions of washed, nonpigmented, nonproliferating Serratia marcescens incubating at 27 C induced biosynthesis of prodigiosin. Four amino acids of casein hydrolysate, dl-aspartic acid, l-glutamic acid, l-proline, and l-alanine caused formation of pigment when added individually. dl-Ornithine also was effective. Optimal concentrations for maximal pigmentation were 5 to 10 mg/ml; at these high concentrations, d-serine also induced biosynthesis of some prodigiosin. dl-Alanine and -ornithine were as effective as the l-iosomers, but l-glutamic acid and l-proline gave better responses than their racemic mixtures. Kinetics of prodigiosin biosynthesis after addition of dl-alanine (20 mg/ml) were similar to those of cells suspended in 0.2% casein hydrolysate. The other amino acids were less effective. Addition of 5 mg of dl-alanine or casein hydrolysate per ml to minimal medium increased by 30% the amount of prodigiosin formed by growing cells after incubation for 7 days at 27 C. Cultures grown for 7 days at 27 C in 0.2% casein hydrolsate formed more prodigiosin than did suspensions of nonproliferating cells containing individual amino acids or casein hydrolysate. However, more pigment was produced by cells suspended in l-alanine (5 mg/ml) or l-proline (10 mg/ml) than when suspended in 0.4% natural or synthetic casein hydrolysate. Filtrates from suspensions of nonproliferating cells forming pigment in l-proline induced more rapid formation of prodigiosin, but filtrates from suspensions in dl-alanine did not. The data supported the hypothesis that pyrrole groups of prodigiosin may be synthesized from 5-carbon amino acids such as proline, ornithine, aspartic, and glutamic acids, but the role of alanine is unknown.

Alanine↗
PubMed1971

Influence of temperature of incubation and type of growth medium on pigmentation in Serratia marcescens.

Maximal amounts of prodigiosin were synthesized in either minimal or complete medium after incubation of cultures at 27 C for 7 days. Biosynthesis of prodigiosin began earlier and the range of temperature for formation was greater in complete medium. No prodigiosin was formed in either medium when cultures were incubated at 38 C; however, after a shift to 27 C, pigmentation ensued, provided the period of incubation at 38 C was not longer than 36 hr for minimal medium or 48 hr for complete medium. Washed, nonpigmented cells grown in either medium at 38 C for 72 hr could synthesize prodigiosin when suspended in saline at 27 C when casein hydrolysate was added. These suspensions produced less prodigiosin at a slower rate than did cultures growing in casein hydrolysate at 27 C without prior incubation at 38 C. Optimal concentration of casein hydrolysate for pigment formation by suspensions was 0.4%; optimal temperature was 27 C. Anaerobic incubation, shift back to 38 C, killing cells by heating, or chloramphenicol (25 mug/ml) inhibited pigmentation. Suspensions of washed cells forming pigment reached pH 8.0 to 8.3 rapidly and maintained this pH throughout incubation for 7 days. Measurements of viable count and of protein, plus other data, indicated that cellular multiplication did not occur in suspensions of washed cells during pigment formation. By this procedure utilizing a shift down in temperature, biosynthesis of prodigiosin by washed cells could be separated from multiplication of bacteria.

Anti-Bacterial Agents↗
PubMed1963

SELECTIVE INHIBITION OF PROLINE-INDUCED PIGMENTATION IN WASHED CELLS OF SERRATIA MARCESCENS.

Blizzard, John L. (University of Houston, Houston, Texas) and G. E. Peterson. Selective inhibition of proline-induced pigmentation in washed cells of Serratia marcescens. J. Bacteriol. 85:1136-1140. 1963.-Streptomycin, chloramphenicol, and tetracyclines inhibited the synthesis of prodigiosin by Serratia marcescens strain D1. This occurred at concentrations of the antibiotic too low to inhibit the growth of the organism in either agar media or broth cultures. Nonpigmented cells were produced in broth by either adding streptomycin or incubating at 37 C. After being washed and resuspended in aqueous saline containing either casein hydrolysate, l-proline, or a glycine-succinate mixture and incubated at 27 C for 24 hr, these cells formed pigment. The appearance of pigment was preceded by a lag period of 10 hr. Prodigiosin production by these washed suspensions of cells was completely inhibited by either streptomycin or glucose, or by incubation at 37 C instead of 27 C. Even though pigmentation by washed-cell suspensions was induced by proline, the utilization of proline was not affected by streptomycin or glucose, or by incubation at 37 C. To block pigmentation completely, streptomycin had to be added to proline-supplemented cells before they were 10 hr old. Addition of the antibiotic after the end of the induction period caused either partial or no inhibition of pigment production. Streptomycin caused an increase in the endogenous respiration of S. marcescens but failed to affect the constitutive enzymes that oxidize glucose. The possible relationships of these phenomena are discussed. Weil (1952) reported that low concentrations of chloramphenicol and certain tetracyclines inhibit the synthesis of prodigiosin while permitting growth by Serratia marcescens. He noted the potential value to "mode-of-action" studies of an organism having certain functions selectively inhibited by antibiotics. We confirmed Weil's (1952) observations and found that streptomycin at low concentration would also inhibit the synthesis of prodigiosin without impeding growth. Further studies of the selective inhibition of prodigiosin synthesis by streptomycin were performed using nonproliferating suspensions of washed cells (Gott and Williams, 1961). Either a glycine-succinate mixture or l-proline could cause nonproliferating cells to form pigment. A period of induction preceded the formation of pigment. Either streptomycin or glucose, or an incubation temperature of 37 C, inhibited the proline-induced pigmentation by washed cells. Further investigations provided insights to these findings.

Amino Acids↗
PubMed1975

Toxigenic studies with the antibiotic pigments from Serratia marcescens.

Prodigiosin, obtained from the bacterium, Serratia marcescens, was extracted in five organic solvents, petroleum ether, chloroform, acetone, ethanol, and methanol, and the fractions were labeled PE-1, C-2, A-3, E-4, and M-5 respectively. The effects of prodigiosin and its fractions on embryogenesis showed the whole pigment and C-2 fraction to be highly toxigenic while other fractions demonstrated toxicities approaching LD50 values of 26-30 mug/egg when dissolved in 100% dimethyl sulfoxide. The E-4 fraction in DMSO was least toxic. Ninety-five percent ethanol proved to be highly toxic at a dose level of 0.1 ml/egg indicating that it was an unsuitable solvent for studies of this nature. Disc-agar diffusion sensitivity studies were performed against E. coli, E. aerogenes, S. aureus, B. subtilis, and P. aeruginosa with prodigiosin and fractions dissolved in 100% DMSO. The solvent was found to have no diffusible bacteriostatic activity in vitro. However, prodigiosin and the ethanol (E-4) and methanol (M-5) fractions produced inhibition zones with every organism tested. Data presented below indicate that prodigiosin extracts have toxigenic effects on chick embryos and inhibit the growth of several species of bacteria.

Acetone↗
PubMed2005

Characterization of bacterium isolated from the sediment at coastal area of Omura Bay in Japan and several biological activities of pigment produced by this isolate.

Recently we discovered a bacterial strain (MS-02-063) that produces large amounts of red pigment from coastal area of Nagasaki Prefecture, Japan. Comparative 16S rDNA gene sequencing analysis revealed that strain MS-02-063 was phylogenetically closely related to gamma-proteobacterium Hahella sp. MBIC 3957 that produces prodigiosin. However, some physiological and biochemical differences between strain MS-02-063 and Hahella sp. MBIC 3957 were observed. The red pigment (RP-063) produced by this isolate was highly purified from the culture supernatant. It was speculated that RP-063 might be prodigiosin-like pigment in physical properties and biological activities such as antibacterial and cytotoxic activity. Antibacterial activity of RP-063 was examined by an agar dilution method. The results indicated that RP-063 showed antibacterial activity for specific for pathogenic gram-positive bacteria such as Staphylococcus aureus. The potency of antibacterial activity against S. aureus was nearly equal to those of tetracycline. Moreover, RP-063 showed inhibition of the superoxide generation by 12-O-tetradecanoylphorbol-13-acetate (TPA)-stimulated mouse macrophage RAW 264.7 cell line. Prodigiosin members have a wide variety of biological properties, including anticancer and antimalarial, etc. Especially, potent immunosuppressive properties have been reported for prodigiosin members with the mechanism of action different from that of the other well known immunosuppressors in atopic dermatitis therapy such as cyclosporin A, FK506 and rapamycin. It is suggested that RP-063 may be able to arrest the inflammation caused by superantigens secreted from S. aureus, which colonized skin on atopic dermatitis as well as suppression of activated lymphocyte proliferation and superoxide generation from leucocytes.

Animals↗
PubMed1994

Effect of various growth conditions on pigmentation of Serratia marcescens.

Prodigiosin biosynthesis in Serratia marcescens depends on growth conditions. In the presence of various concentrations of NaCl, pigment synthesis started later, but up to 4-5% NaCl increased prodigiosin accumulation per biomass unit. Visible light (< 2,000 lux) influenced pigmentation without changing the growth characteristics of the culture. The maximum prodigiosin content in dark and light cultures was observed on days 3-4 and 2-3, respectively. The influence of illumination conditions on pigmentation of S. marcescens was demonstrated. Light directly affects the pigment itself which is synthesized by the culture. Growing cells contain mono and dimer forms of prodigiosin in glycerol, with light influencing these pigments.

Culture Media↗
PubMedDate not supplied

Isolation of Serratia marcescens in the midgut of Rhodnius prolixus: impact on the establishment of the parasite Trypanosoma cruzi in the vector.

The effects of resident bacteria in the stomach of 5th-instar larvae of Rhodnius prolixus on the erythrocyte lysis and Trypanosoma cruzi infection were studied. The bacteria population increased approximately 10,000-fold after feeding. Hemolysis rose to approximately 28% within 24h postfeeding and then linearly grew until day 4 attaining almost 100%. The number of surviving Y strain of T. cruzi in the stomach declined drastically, while the infection with Dm28c clone was maintained stable. Five days after feeding, few different types of bacterial colonies were obtained when stomach content suspensions were spread onto BHI agar plates. The hemolytic bacteria were isolated and identified as Serratia marcescens biotype A1a (referenced as RPH), which produces the pigment prodigiosin. In vitro experiments, comparing incubations of RPH with S. marcescens SM365, a prodigiosin pigment producer, and S. marcescens DB11, a nonpigment variant, as a control, with erythrocytes and T. cruzi demonstrated that: (i) at 30 degrees C, SM365 and RPH diminished the populations of Y strain, but not DM28c clone, and DB11 was unable to lyse both T. cruzi strains; (ii) at 0 degrees C, SM263 and RPH killed the flagellates, but DB11 did not; and (iii) all three strains of S. marcescens were able to lyse erythrocytes. These results suggest that S. marcescens trypanolytic activity from the SM365 and RPH strains is distinct from the hemolytic activity and that prodigiosin is an important factor for the trypanolytic action of the bacteria.

Animals↗
PubMedDate not supplied

[The effect of the cultivation conditions on the growth and pigmentation of Serratia marcescens].

The influence of cultivation conditions on the growth and pigmentation of S. marcescens was studied. The cultures under study grew in media containing glycerin, glucose or acetate and organic or mineral nitrogen. Pigment formation occurred in a medium with organic nitrogen and glycerin or acetate, but not glucose. Sodium chloride inhibited the growth of cultures, but at a concentration not exceeding 4-5% increased the accumulation of prodigiosin. Prodigiosin was accumulation by the culture growing at 28 degrees C, while at 37 degrees C no accumulation of the pigment occurred. The illumination of the growing culture with visible light decreased the intensity of its pigmentation. Prodigiosin apparently plays an important role in the metabolism of S. marcescens and is linked with the energy exchange of the cell.

Anti-Bacterial Agents↗
PubMedDate not supplied

[Proposed neotype Streptomyces ruber (Krainsky, 1914) Waksman et Henrici, 1948].

Culture 78 was proposed as a neotype of Streptomyces ruber. It was isolated from the soils of the Baikal region and was closest, in its taxonomic properties, to the original description of the species [13] whose representative had been lost. Cultures from different microbial collections designated as S. ruber were shown to be unlike the original description. The neotype had the following taxononic properties: the cell wall of type I; spiral sporophores with extended spirals having 2-3 coils; oval spores with a smooth envelope; greyish pink aerial and dark-red substrate mycelia; a red pigment not passing into the medium; slow gelatin liquefaction and milk peptonization; weak starch hydrolysis; assimilation of glucose, xylose, rammose, fructose, and inositol; weak growth on arabinose, raffinose and mannitol, but not on sucrose; no formation of melanoid pigments; synthesis of riboflavin and prodigiosin pigments; inhibition of Gram-positive bacterial and acid-resistant mycobacterial growth; no inhibition of yeast and fungal growth. The culture was sensitive to streptomycin, neomycin, gentamycin, monomycin, tetracycline,erythromycin, oleandomycin, lincomycin, ristomycin, levomycetin, polymyxin and fusidin, but resistant in penicillin. The population was composed of six variants [3]: main, faded, asporogenic red, asporogenic yellow, asporogenic white and nocardia-like. The latter two were not capable of riboflavin and prodigiosin formation. The asporogenic yellow variant was a monosynthetic organism: it formed riboflavin, but could not synthesize prodigiosin. The neotype of S. ruber 78 is deposited withthe national Collection of Microorganisms (the reference number is VKM A-611).

Anti-Bacterial Agents↗