Effect of physical parameters in enhancing prodigiosin production and anti-MRSA activity of marine bacterium, Serratia marcescens IBRL USM84

Aims@#Marine bacteria have been reported to produce potential natural pigment with pharmaceutical properties and their growth can be manipulated in the laboratory to increase pigment production and their antimicrobial activity. Hence, this study aimed to enhance the prodigiosin production in Serratia marcescens IBRL USM84 by improving physical conditions.@*Methodology and results@#The quantification of the pigment produced by S. marcescens IBRL USM84, bacterial cell growth, and its antibacterial activity in the broth medium were determined using a spectrophotometry method. Meanwhile, the antibacterial effect of red pigment on MRSA cells was observed under a scanning electron microscope (SEM). This marine isolate produced the highest yield of prodigiosin (6.95 μg/mL) when cultivated in marine broth with the addition of 0.2% of agar, 25 °C incubation temperature, initial medium pH of 7, 150 rpm of agitation speed for 48 h of cultivation time under light illumination. There was an increment of 151.81% in prodigiosin production after enhancement compared to before the enhancement of cultural conditions. SEM observations revealed that severe damage to the cell’s morphologies was exposed to red pigment as indicated by the formation of small dents, which led to completely collapse and eventually, cell death.@*Conclusion, significance and impact of study@#A positive correlation between pigment production and antibacterial activity was observed in the present study. The results supported the fact that marine bacteria are a reservoir of various pigments with antimicrobial properties. Also, the pigment production by S. marcescens and its antibacterial activity were significantly influenced by physical parameters.

Prodigiosin from Serratia: Synthesis and potential applications

Prodigiosin is a red pigment with a pyrrolylpyrromethane skeleton. It is mainly produced by bacterial strains belonging to the Serratia genus, but also by some other genera, including Streptomyces and Vibrio. Within the genus Serratia, the pigment is generally produced as a virulence factor. However, it also has many important beneficial biological activities such as immunosuppressive and anti- proliferative activities. Moreover, the pigment has many industrial applications in textile and cosmetics. In this mini-review, we discuss the genetic and molecular mechanisms supporting prodigiosin synthesis and production from the Serratia genus, as well as its potential applications.

Prodigiosin from Serratia: Synthesis and potential applications

Prodigiosin is a red pigment with a pyrrolylpyrromethane skeleton. It is mainly produced by bacterial strains belonging to the Serratia genus, but also by some other genera, including Streptomyces and Vibrio. Within the genus Serratia, the pigment is generally produced as a virulence factor. However, it also has many important beneficial biological activities such as immunosuppressive and anti- proliferative activities. Moreover, the pigment has many industrial applications in textile and cosmetics. In this mini-review, we discuss the genetic and molecular mechanisms supporting prodigiosin synthesis and production from the Serratia genus, as well as its potential applications.

Enhanced production of prodigiosin by Serratia marcescens FZSF02 in the form of pigment pellets

Background: Prodigiosin has been demonstrated to be an important candidate in investigating anticancer drugs and in many other applications in recent years. However, industrial production of prodigiosin has not been achieved. In this study, we found a prodigiosin-producing strain, Serratia marcescens FZSF02, and its fermentation strategies were studied to achieve the maximum yield of prodigiosin. Results: When the culture medium consisted of 16.97 g/L of peanut powder, 16.02 g/L of beef extract, and 11.29 mL/L of olive oil, prodigiosin reached a yield of 13.622 ± 236 mg/L after culturing at 26 °C for 72 h. Furthermore, when 10 mL/L olive oil was added to the fermentation broth at the 24th hour of fermentation, the maximum prodigiosin production of 15,420.9 mg/L was obtained, which was 9.3-fold higher than the initial level before medium optimization. More than 60% of the prodigiosin produced with this optimized fermentation strategy was in the form of pigment pellets. To the best of our knowledge, this is the first report on this phenomenon of pigment pellet formation, which made it much easier to extract prodigiosin at low cost. Prodigiosin was then purified and identified by absorption spectroscopy, HPLC, and LCMS. Purified prodigiosin obtained in this study showed anticancer activity in separate experiments on several human cell cultures: A549, K562, HL60, HepG2, and HCT116. Conclusions: This is a promising strain for producing prodigiosin. The prodigiosin has potential in anticancer medicine studies.

Antibacterial pigment production by Serratia marcescens using different casein types obtained from milk / Producción de un pigmento antibacteriano con Serratia marcescens usando diferentes tipos de caseína obtenida de leche

ABSTRACT Prodigiosin, a pigment produced by Serratia marcescens inhibits the growth of different microorganisms and the proliferation of some human cancer cell lines. Prodigiosin is usually produced by fermentations of substrates such as starch and proteins, and pigment yield depends on the concentration of the carbon source, stirring speed of cultures, temperature and time of incubation, nitrogen sources, and pH of medium. In the present study, pigment production was assessed using two types of casein as substrate; pigment yield was greater when casein precipitated with vinegar was used as substrate than when high purity casein was. The maximum prodigiosin production was achieved with 10 g/L casein at pH 8.0. Additionally, supplementation of culture media with glucose was found to considerably decrease prodigiosin production and growth inhibition of Staphylococcus aureus, which is directly related to pigment yield. Production in stirred-tank bioreactor at 0.75 vvm aeration was higher than that at 0.5 and 1.0 vvm. Substrate type, concentration and pH affected pigment production in Erlenmeyer flasks, whereas aeration rate influenced pigment production in a stirred-tank bioreactor.

RESUMEN La prodigiosina es un pigmento producido por Serratia marcescens que inhibe el crecimiento de diferentes microorganismos y la proliferación de algunas líneas celulares de cáncer humano. La prodigiosina generalmente se produce a través de fermentaciones con sustratos como almidón y proteínas y su producción depende de la concentración de la fuente de carbono, la velocidad de agitación en los cultivos, la temperatura y el tiempo de incubación, las fuentes de nitrógeno y el pH del medio. En este trabajo se evaluó la formación de pigmento usando dos tipos de caseína como sustrato; la producción de pigmento fue mayor cuando se utilizó caseína precipitada con vinagre que cuando se utilizó caseína de alta pureza. La máxima concentración de prodigiosina se logró con 10 g/L de caseína a un pH de 8.0. Se encontró además que suplementar los medios de cultivo con glucosa disminuye considerablemente la producción de prodigiosina y la inhibición de Staphylococcus aureus, la cual está relacionada directamente con la producción de pigmento. La producción en biorreactor de tanque agitado con una aireación de 0.75 vvm fue mayor que con 0.5 y 1.0 vvm. El tipo de sustrato, la concentración y el pH afectaron la producción de pigmento en Erlenmeyer mientras que la velocidad de aireación influyó sobre la producción de pigmento en biorreactor de tanque agitado.

The potential use of prodigiosin as a shrimp feed additive and its dynamic influence on the shrimp gut microbial community – an in vitro gut model

Aims@#A simple in vitro model system was applied in this study assessing the dynamics of the microbial community associated with the shrimp gut system to understand the changes that influence dietary variables. @*Methodology and results@#The diversity and abundance of microbiome were monitored within two different treatment slurries inoculated with shrimp faecal samples as to mimic the effect of diet manipulation, and 16S rRNA gene of MiSeq Illumina-based sequencing was applied. The different diets tested were a commercial standard diet and a prodigiosin added diet. There was very clear separation between the commercial standard diet and prodigiosin added diet as revealed by the total viable counts (TVC) and sequencing data. It suggested that the microbial community of the shrimp gut system exhibited a dynamic response with the treatments and allochthonous bacterial present. The prodigiosin added diet was clearly separated from the commercial standard diet serving as a potential shrimp feed additive. The sequencing data analysis showed that members of the genera Vibrio, Shigella and Photobacterium became predominant on the commercial standard diet treatment. The prodigiosin-added diet treatments indicated an abundance of members of the genera Micrococcus, Arthrobacter, and Shigella. @*Conclusion, significance and impact of study@#In vitro model system-based testing of diets could be a useful method to determine the potential effect of diet manipulation on shrimp gut system microbiome members.

Production, characterization and antibacterial activity of prodigiosin pigment produced by Pseudoalteromonas rubra BF1A IBRL associated with a marine macroalgae Enteromorpha sp

Aims@#Marine bacteria are a great source of natural pigments, which can be used as colouring agent in food, textile, cosmetics and aquaculture industry to overcome the drawbacks poses by the synthetic pigments. The aim of the study is to identify the potential bio pigment producer, determine the antimicrobial susceptibilities, and characterize the pigment produced. @*Methodology and results@#In this study, the surface attached marine bacteria isolated from the surface of seaweed, Enteromorpha sp. has been identified as Pseudoalteromonas rubra BF1A IBRL through the molecular identification step. This species produced intracellular and extracellular red pigment with antibacterial activity. The susceptible bacteria include Bacillus subtilis, Bacillus cereus, Methicillin Resistant Staphylococcus aureus, Staphylococcus aureus and also Acinetobacter anitratus with inhibition zone ranges from 7.33 to 10.33 mm, whereas Minimum inhibitory concentration (MIC) values ranges from 0.055 to 8.88 mg/mL. The UV/vis analysis indicated that the maximal absorbance of ISO and DE pigment extract were at 531 and 534 nm, respectively. Based on the antimicrobial activity, the extracellular extract poses greater antibacterial activity, thus was selected as the potential pigment extract and were further evaluated. The Thin Layer Chromatography (TLC) profile of the DE extract showed one major band under visible light ((Rf = 0.87) and the bioautography analysis of the pigmented band showed positive activity against both Gram-positive and Gram-negative bacteria. The pigment in DE extract was identified as prodigiosin based on the spectroscopic properties, presumptive test and HPLC analysis. @*Conclusion, significance and impact of study@#This study highlights the dual benefits of the P. rubra BF1A IBRL pigment extract, which exhibited both tinctorial and pharmacology benefits, thus it can be act as colouring agent with own preservative value in food, textile, or cosmetics industries.

Assessment of cultivation parameters influencing the growth, pigment production and anti-MRSA activity of Pseudoalteromonas rubra BF1A IBRL isolated from Malaysian marine environment

Aims@#Pigments are coloured substances that exhibit important characteristics to many industries including food, textile, cosmetics, food, pharmaceutical and also aquaculture industry. Naturally derived pigments from marine bacteria do not only exhibit the tinctorial property but are also known to possess broad range of antimicrobial activities. From the industrial point of view, the necessity to obtain suitable culture conditions for maximum yield of cell growth and pigment production is of utmost importance. @*Methodology and results@#The effect of cultural conditions, including light, pH, temperature, agitation speed and size of inoculum on bioactivity of an epiphytic marine bacteria, Pseudoalteromonas rubra BF1A IBRL was studied using shake flask technology. The antimicrobial activity was determined using the Lorian method. As a result, prodigiosin pigment extract obtained from P. rubra BF1A IBRL showed inhibitory activity against the MRSA strain. Pseudoalteromonas rubra BF1A IBRL produced the highest level of prodigiosin and anti-MRSA activity (P<0.05) in Marine broth at initial pH of 7.6 incubated at dark condition at temperature of 26 °C, agitation speed of 120 rpm and 2% (v/v) (1 × 106 CFU/mL) of inoculums size. @*Conclusion, significance and impact of study@#A high correlation between pigmentation and antibacterial activity were observed anticipating that the pigment has its own antibacterial properties. The above findings supported the fact that epiphytic marine bacteria were fruitful source for pigmented bioactive compounds, and the physical parameters had significantly influence of the pigment production.

Assessment of cultivation parameters influencing the growth, pigment production and anti-MRSA activity of Pseudoalteromonas rubra BF1A IBRL isolated from Malaysian marine environment

Aims@#Pigments are coloured substances that exhibit important characteristics to many industries including food, textile, cosmetics, food, pharmaceutical and also aquaculture industry. Naturally derived pigments from marine bacteria do not only exhibit the tinctorial property but are also known to possess broad range of antimicrobial activities. From the industrial point of view, the necessity to obtain suitable culture conditions for maximum yield of cell growth and pigment production is of utmost importance. @*Methodology and results@#The effect of cultural conditions, including light, pH, temperature, agitation speed and size of inoculum on bioactivity of an epiphytic marine bacteria, Pseudoalteromonas rubra BF1A IBRL was studied using shake flask technology. The antimicrobial activity was determined using the Lorian method. As a result, prodigiosin pigment extract obtained from P. rubra BF1A IBRL showed inhibitory activity against the MRSA strain. Pseudoalteromonas rubra BF1A IBRL produced the highest level of prodigiosin and anti-MRSA activity (P<0.05) in Marine broth at initial pH of 7.6 incubated at dark condition at temperature of 26 °C, agitation speed of 120 rpm and 2% (v/v) (1 × 106 CFU/mL) of inoculums size. @*Conclusion, significance and impact of study@#A high correlation between pigmentation and antibacterial activity were observed anticipating that the pigment has its own antibacterial properties. The above findings supported the fact that epiphytic marine bacteria were fruitful source for pigmented bioactive compounds, and the physical parameters had significantly influence of the pigment production.

Using prodigiosin against some gram-positive and gram-negative bacteria and Trypanosoma cruzi

This work aimed to explore the action of natural prodigiosin on both bacterial organisms and Trypanosoma cruzi cells. Methods: Natural prodigiosin pigment was extracted and purified from cultures of Serratia marcescens. Two media, peanut broth and peptone glycerol broth, both recommended in the literature for prodigiosin production, were compared. The prodigiosin obtained was employed to explore its antimicrobial properties against both bacteria and Trypanosoma cruzi cells. Results: Peanut broth yielded four times more prodigiosin. The prodigiosin showed remarkable activity (minimal inhibitory concentrations in the range of 2-8 µM for bacteria and half maximal inhibitory concentration of 0.6 µM for Trypanosoma cruzi). In fact, the prodigiosin concentration required to inhibit parasite growth was as low as 0.25 mg/l versus 4.9 mg/l of benznidazole required. Furthermore, atomic force microscopy revealed marked morphological alterations in treated epimastigote forms, although no pore-formation activity was detected in protein-free environments. Conclusions: This work demonstrates the potential usefulness of prodigiosin against some gram-positive and gram-negative bacteria and Trypanosoma cruzi although further studies must be done in order to assess its value as a candidate molecule.(AU)

Effect of prodigiosin on the alleviation of the intestinal inflammation of weaned rats based on 1H-NMR spectroscopy study and biochemistry indexes

Weaning results in intestinal dysfunction, mucosal atrophy, transient anorexia, and intestinal barrier defects. In this study, the effect of prodigiosin (PG) on the intestinal inflammation of weaned rats was investigated by using 1H-NMR spectroscopy and biochemistry indexes to regulate the intestinal metabolism. After administration for 14 days, the body mass of the PG group was increased by 1.29- and 1.26-fold compared with those of the control and alcohol groups, respectively, using a dose of 200 µg PG·kg-1 body weight per day. PG increased organic acid content and decreased moisture, pH values, and free ammonia in feces. In addition, PG alleviated the intestinal inflammation of weaned rats. The analysis of 1H-NMR signal peak attribution and the model validation of metabolic data of feces contents showed that PG significantly affected the metabolism of small molecular compounds in the intestinal tract of weaned rats. This study presents the promising alternative of using PG to alleviate intestinal inflammation effectively in the intestinal tract of weaned rats.

Progress in microbial production of prodigiosin / 生物工程学报

Prodigiosin is an important natural red pigment that is produced as a secondary metabolite by microorganisms, and has great potential applications in the field of pharmaceutical development, environmental management and dye preparation. This paper reviews recent research progress in the production of prodigiosin by microbial fermentation, including discovery and modification of the prodigiosin-producing microorganisms, regulation and optimization of prodigiosin fermentation and extraction process, and resolution of biosynthetic pathway of prodigiosin and related transcriptional regulation. Finally, we discussed the future research directions in microbial production of prodigiosin.

Enhanced production of prodigiosin by Serratia marcescens MO-1 using ram horn peptone

This work addresses the production of prodigiosin from ram horn peptone (RHP) using MO-1, a local isolate in submerged culture. First, a novel gram-negative and rod-shaped bacterial strain, MO-1, was isolated from the body of the grasshopper (Poecilemon tauricola Ramme 1951), which was collected from pesticide-contaminated fields. Sequence analysis of 16S rDNA classified the microbe as Serratia marcescens. The substrate utilization potential (BIOLOG) and fatty acid methyl ester profile (FAME) of S. marcescens were also determined. The effect of RHP on the production of prodigiosin by S. marcescens MO-1 was investigated, and the results showed that RHP supplementation promoted the growth of MO-1 and increased the production of prodigiosin. A concentration of 0.4% (w/v) RHP resulted in the greatest yield of prodigiosin (277.74 mg/L) after 48 h when mannitol was used as the sole source of carbon. The pigment yield was also influenced by the types of carbon sources and peptones. As a result, RHP was demonstrated to be a suitable substrate for prodigiosin production. These results revealed that prodigiosin could be produced efficiently by S. marcescens using RHP.

Prodigiosin - an antibacterial red pigment produced by Serratia marcescens IBRL USM 84 associated with a marine sponge Xestospongia testudinaria.

The objectives of present study are to extract the antibiotic compound from marine isolate and to determine its in vitro antimicrobial activity against bacteria. A marine bacterial isolate Serratia marcescens IBRL USM 84 was isolated from the surface of a marine sponge Xestospongia testudinaria. This species of bacteria produced red pigment with antibacterial activity. The red antibacterial pigment was produced intracellularly and inhibited 13 out of 18 tested bacteria, with Gram positive was more susceptible than the Gram negative bacteria. The growth and antibacterial red pigment production profiles demonstrated the highest antibacterial red pigment production was achieved at the 48 hours of cultivation (14.08 U/ml) time in marine broth when incubated at 25 °C with 150 rpm agitation. The antibacterial red pigment was extracted, purified and confirmed as prodigiosin.

Studies on the Prodigiosin Production from Streptomyces coelicolor in Liquid Media by Using Heated Lactobacillus rhamnosus.

Prodigiosin (PG) is not only an antibiotic but it has also been characterized immunosuppressive and anticancer activities. Prodigiosin is produced by Streptomyces coelicolor and Seratia marcescens. In this study, prodigiosin (PG) production of S. coelicolor was optimized using 50μL, 100μL of heated cells at 70oC and 100oC of Lactobacillus rhamnosus. With 50μL heat-killed cells at 70oC L. rhamnosus, prodigiosin (PG) production of S. coelicolor enhanced the maximum prodigiosin concentration (9.79  1.68 mg/L) by six-fold at the early stage in the second day incubation in the comparison with prodigiosin (PG) production of S. coelicolor without heated L. rhamnosus (1.43  0.22 mg/L). There were the affects of the components of L. rhamnosus on PG production. This is the first report that L. rhamnosus affecting on PG production in S. coelicolor.

Effect of certain natural products and organic solvents on quorum sensing in Chromobacterium violaceum

OBJECTIVE@#To investigate the effect of seed extracts of Pongamia pinnata, Pyrus pyrifolia, and Manilkara hexandra, bacterial pigment prodigiosin, and three organic solvents (ethanol, methanol, and dimethylsulfoxide), on quorum sensing (QS) in Chromobacterium violaceum (C. violaceum).@*METHODS@#C. violaceum was challenged with plant extracts prepared by microwave assisted extraction method, prodigiosin, and organic solvents. Effect of these test substances on C. violaceum growth, and quorum sensing regulated pigment (violacein) production was studied by broth dilution assay. High performance liquid chromatography was also applied to generate chromatographic fingerprint of the active extracts. Effect of sub-minimum inhibitory concentration level of the antibiotic streptomycin on quorum sensing regulated pigment production was also studied.@*RESULTS@#Pongamia pinnata seed extracts and prodigiosin were found to possess anti-QS, and Manilkara hexandra and Pyrus pyrifolia seed extracts to possess QS-enhancing effect in C. violaceum. Dimethylsulfoxide was found to enhance violacein production, whereas ethanol and methanol reduced violacein production in C. violaceum. Streptomycin at sub-minimum inhibitory concentration level was able to significantly arrest QS-regulated pigment production in C. violaceum and Serratia marcescens.@*CONCLUSIONS@#Prodigiosin and the seed extracts used in this study could affect quorum sensing in C. violaceum to a notable extent. Results of this study also emphasize the importance of inclusion of appropriate solvent controls (negative controls) in bioassays designed for screening of antimicrobial and/or anti-QS compounds. Antipathogenic potential of low concentrations of streptomycin was also demonstrated.

Effect of certain natural products and organic solvents on quorum sensing in Chromobacterium violaceum

Objective: To investigate the effect of seed extracts of Pongamia pinnata, Pyrus pyrifolia, and Manilkara hexandra, bacterial pigment prodigiosin, and three organic solvents (ethanol, methanol, and dimethylsulfoxide), on quorum sensing (QS) in Chromobacterium violaceum (C. violaceum). Methods: C. violaceum was challenged with plant extracts prepared by microwave assisted extraction method, prodigiosin, and organic solvents. Effect of these test substances on C. violaceum growth, and quorum sensing regulated pigment (violacein) production was studied by broth dilution assay. High performance liquid chromatography was also applied to generate chromatographic fingerprint of the active extracts. Effect of sub-minimum inhibitory concentration level of the antibiotic streptomycin on quorum sensing regulated pigment production was also studied. Results: Pongamia pinnata seed extracts and prodigiosin were found to possess anti-QS, and Manilkara hexandra and Pyrus pyrifolia seed extracts to possess QS-enhancing effect in C. violaceum. Dimethylsulfoxide was found to enhance violacein production, whereas ethanol and methanol reduced violacein production in C. violaceum. Streptomycin at sub-minimum inhibitory concentration level was able to significantly arrest QS-regulated pigment production in C. violaceum and Serratia marcescens. Conclusions: Prodigiosin and the seed extracts used in this study could affect quorum sensing in C. violaceum to a notable extent. Results of this study also emphasize the importance of inclusion of appropriate solvent controls (negative controls) in bioassays designed for screening of antimicrobial and/or anti-QS compounds. Antipathogenic potential of low concentrations of streptomycin was also demonstrated.

Enhanced production of prodigiosin production in serratia marcescens.

Prodigiosins, a family of natural red pigments characterized by a common Pyrrolylpyrromethane skeleton are produced by various bacteria that first characterized from Serratia marcescens, Pseudomonas magneslorubra, Vibrio psychroerythrous etc. In this study, different media like nutrient broth, peptone glycerol broth, powdered peanut seed broth and powdered sesame seed broth, sugar substrates (maltose and glucose) and different oils like peanut oil, sesame oil, coconut oil at different temperature were used to study the prodigiosin production in Serratia marcescens. The highest pigment production was observed in powdered peanut seed broth (37.6mg/ml) and powdered sesame seed broth (16.5mg/ml) when compared to the use of nutrient broth (0.51mg/ml) and peptone glycerol broth (0.3 mg/ml) at 280c, Nutrient broth with maltose yielded (1.82mg/ml) of prodigiosin. Among different oils, peanut oil showed high pigment production (2.72 mg/ml) and these pigment can be used to dye cotton which showed good color tone.

Assessment of process parameters influencing the enhanced production of prodigiosin from Serratia marcescens and evaluation of its antimicrobial,antioxidant and dyeing potentials

Aims: Prodigiosin is a bright red pigment produced by certain strains of Serratia marcescens, characterized by a common pyrrolylpyrromethane skeleton. This pigment is found to possess antibacterial, antifungal, immunosuppressive and antiproliferative activity. The present study aimed at designing process parameters for the enhanced production of this pigment. Methodology and Results: Peptone glycerol broth was selected as the best synthetic medium. The effects of various media components and process parameters like carbon and nitrogen sources, temperature, pH, incubation period and other supplements were investigated. Maximal amount of prodigiosin was produced at temperature 25 °C, pH 7.0 and incubation period of 48 h. Supplementation of media with maltose and peptone yielded maximal amount of prodigiosin. Incorporation of minimal amount of supplements like silica gel, iron salts, inorganic phosphate also showed promising results. Chromatographic separations suggested that prodigiosin is made up of three different fractions (purple, orange and red). Further investigation of antimicrobial properties of prodigiosin revealed that it is a potent inhibitor against gram positive bacteria like Staphylococcus aureus and Bacillus cereus and fungal pathogens like Candida albicans, C. parapsilosis and Cryptococcus sp. This antimicrobial potency remained stable under a wide range of temperature and pH. The antioxidant capacity of prodigiosin was found to be 22.05 Bg ascorbic acid equivalents/ml of extract. When applied to textiles, prodigiosin resisted the action of acid, alkali and detergent. Conclusion, Significance and Impact of study: Besides combating gram positive bacterial pathogens and some pathogenic yeasts, prodigiosin with strong dyeing and antioxidant activity may find broad applications in textile and therapeutic industries.

SUDIES ON THE CHEMICAL CONSTITUENTS OF THE DAYAWAN BAY GERM PSEUDOMONAS SP / 中国海洋药物

A red pigmant, procligiosin, have been isolated from the metabolite of germ pseudomonas sp. collected from the Dayawan Bay. Its structure was established on the basis of the spectral method.