Improving Bioprocess Conditions for the Production of Prodigiosin Using a Marine Serratia rubidaea Strain.

The enormous potential attributed to prodigiosin regarding its applicability as a natural pigment and pharmaceutical agent justifies the development of sound bioprocesses for its production. Using a Serratia rubidaea strain isolated from a shallow-water hydrothermal vent, optimization of the growth medium composition was carried out. After medium development, the bacterium temperature, light and oxygen needs were studied, as was growth inhibition by product concentration. The implemented changes led to a 13-fold increase in prodigiosin production in a shake flask, reaching 19.7 mg/L. The conditions allowing the highest bacterial cell growth and prodigiosin production were also tested with another marine strain: S. marcescens isolated from a tide rock pool was able to produce 15.8 mg/L of prodigiosin. The bioprocess with S. rubidaea was scaled up from 0.1 L shake flasks to 2 L bioreactors using the maintenance of the oxygen mass transfer coefficient (kLa) as the scale-up criterion. The implemented parameters in the bioreactor led to an 8-fold increase in product per biomass yield and to a final concentration of 293.1 mg/L of prodigiosin in 24 h.

Biosorption of heavy metals from phosphate-processing effluent by Serratia rubidaea NCTC12971 immobilized in Ca-alginate beads.

In this study, the biosorption ability of various potentially toxic elements from phosphate-processing effluent (PPE) using the indigenous bacterium Serratia rubidaea NCTC12971 immobilized in Ca-alginate beads was investigated. The experimental data analyzed by the Langmuir isotherm revealed that the optimum dose of 2 g·100 ml-1 of immobilized S. rubidaea NCTC12971 at pH 7 and a contact time of 48 h allowed the removal of 92.07%, 98.05%, 95.57%, and 88.39% of lead (Pb (II)), cadmium (Cd (II)), copper (Cu (II)), and zinc (Zn (II)), respectively. Moreover, under the Langmuir isotherm, the maximum single-layer adsorption capacity (qmax) of the biosorbent was estimated to 32.14 mg g-1, 45.87 mg g-1, 0.06 mg g-1, and 3.01 mg g-1 for Pb (II), Cd (II), Cu (II), and Zn (II), respectively, under the stated conditions. Alternatively, the regeneration and reuse of the Ca-alginate beads was evaluated. Indeed, after four consecutive adsorption-desorption cycles, there was no significant loss in the biosorption capacity. The effectiveness of the bacterial biosorption as treatment process was evaluated by assessing the phytotoxicity of the treated effluent (TE) on Medicago sativa and Lactuca sativa seed germination and their root elongation. Results exhibited a significant toxicity removal expressed by a notable increase in the germination indices (GI), which reach 80% and 70%, respectively, for Medicago sativa and Lactuca sativa compared to the GI values of 46.6% and 16.6% of the same species in presence of the untreated effluent (PPE).

Isolation and Characterization of a Serratia rubidaea from a Shallow Water Hydrothermal Vent.

Microbial life present in the marine environment has to be able to adapt to rapidly changing and often extreme conditions. This makes these organisms a putative source of commercially interesting compounds since adaptation provides different biochemical routes from those found in their terrestrial counterparts. In this work, the goal was the identification of a marine bacterium isolated from a sample taken at a shallow water hydrothermal vent and of its red product. Genomic, lipidomic, and biochemical approaches were used simultaneously, and the bacterium was identified as Serratia rubidaea. A high-throughput screening strategy was used to assess the best physico-chemical conditions permitting both cell growth and production of the red product. The fatty acid composition of the microbial cells was studied to assess adaptation at the lipid level under stressful conditions, whilst several state-of-the-art techniques, such as DSC, FTIR, NMR, and Ultra-High Resolution Qq-Time-of-Flight mass spectrometry, were used to characterize the structure of the pigment. We hypothesize that the pigment, which could be produced by the cells up to 62 °C, is prodigiosin linked to an aliphatic compound that acts as an anchor to keep it close to the cells in the marine environment.

Production optimization, characterization, and covalent immobilization of a thermophilic Serratia rubidaea lipase isolated from an Algerian oil waste.

A new thermophilic non-induced lipase producer named Serratia rubidaea strain Nehal-mou was isolated from oil waste in Tissemsilat, Algeria. The most influential lipase production parameters were screened by the Plackett-Burman design for enhancing enzyme yield. An optimum condition of a 1.5% of glucose, a 0.01% of potassium, and a 0.025% of manganese contents resulted in a 41.13 U/mL. This yield was 6.29 times higher than the one achieved before the application of the Box-Behnken Design. Lipase activity showed a high organic solvent tolerance following its exposure to hexane, ethanol, methanol, and acetone. Lipase was also perfectly stable in the presence of 10 mM Fe2+, K+, and Na+ ions with more than 75% of the retaining activity. The enzyme half-life times were 22 h, 90 min, and 25 min at 50, 60, and 70 °C respectively. Polyvinyl alcohol (PVA)/boric acid/Starch/CaCO3 were utilized as a carrier for lipase covalent immobilization in order to be used efficiently. The Scanning Electron Microscopy (SEM) Technique and the Fourier Transform Infrared Spectroscopy (FTIR) Method confirmed the covalent bonding success and the excellent carrier characteristics. Thus, the immobilization yield reached 73.5% and the optimum temperature was shifted from 40 to 65 °C. The immobilized lipase kept 80% of its total activity after 10 cycles and had 3 and 3.2-fold half-lives at 70, and 80 °C respectively compared to the free enzyme.

Chromosome-Encoded Broad-Spectrum Ambler Class A ß-Lactamase RUB-1 from Serratia rubidaea.

Whole-genome sequencing of Serratia rubidaea CIP 103234T revealed a chromosomally located Ambler class A ß-lactamase gene. The gene was cloned, and the ß-lactamase, RUB-1, was characterized. RUB-1 displayed 74% and 73% amino acid sequence identity with the GIL-1 and TEM-1 penicillinases, respectively, and its substrate profile was similar to that of the latter ß-lactamases. Analysis by 5' rapid amplification of cDNA ends revealed promoter sequences highly divergent from the Escherichia coli σ70 consensus sequence. This work further illustrates the heterogeneity of ß-lactamases among Serratia spp.

Investigating the compatibility of the biocontrol agent Clonostachys rosea IK726 with prodigiosin-producing Serratia rubidaea S55 and phenazine-producing Pseudomonas chlororaphis ToZa7.

This study was carried out to assess the compatibility of the biocontrol fungus Clonostachys rosea IK726 with the phenazine-producing Pseudomonas chlororaphis ToZa7 or with the prodigiosin-producing Serratia rubidaea S55 against Fusarium oxysporum f. sp. radicis-lycopersici. The pathogen was inhibited by both strains in vitro, whereas C. rosea displayed high tolerance to S. rubidaea but not to P. chlororaphis. We hypothesized that this could be attributed to the ATP-binding cassette (ABC) proteins. The results of the reverse transcription quantitative PCR showed an induction of seven genes (abcB1, abcB20, abcB26, abcC12, abcC12, abcG8 and abcG25) from subfamilies B, C and G. In planta experiments showed a significant reduction in foot and root rot on tomato plants inoculated with C. rosea and P. chlororaphis. This study demonstrates the potential for combining different biocontrol agents and suggests an involvement of ABC transporters in secondary metabolite tolerance in C. rosea.

Prodigiosin from Serratia rubidaea MJ 24 impedes Helicoverpa armigera development by the dysregulation of Juvenile hormone-dopamine system.

Prodigiosin pigment is a secondary metabolite produced by many bacterial species and is known for its medicinal properties. A few of these prodigiosin-producing bacteria are also reported to be entomopathogenic. It is intriguing to unravel the role of prodigiosin in insecticidal activities and its mode of action. In this study, we have shown the production and characterization of prodigiosin from the Serratia rubidaea MJ 24 isolated from the soil of the Western Ghats, India. Further, we assessed the effect of this pigment on the lepidopteran agricultural pest, Helicoverpa armigera. Prodigiosin-fed H. armigera indicated defective development of insect growth upon treatment. Due to defective early development, about 50% mortality and 40% reduction in body weight were observed in insects fed on a 500 ppm prodigiosin-containing diet. The transcriptomic analysis of these insects indicated significant dysregulation of Juvenile hormone synthesis and response related genes. In addition, dopamine related processes and their resultant melanization and sclerotization processes were also found to be affected. The changes in the expression levels of the key transcripts were further validated using real-time quantitative PCR. The metabolome data confirmed the developmental dysregulation of precursors and products of differentially regulated genes due to prodigiosin. Therefore, the corroborated data suggests that prodigiosin majorly affects H. armigera development through dysregulation of the Juvenile hormone-dopamine system and can be considered as a bioactive scaffold to design insect-pest management compounds. This study provides the first report of in-depth analysis of insecticidal system dynamics in H. armigera insects upon prodigiosin feeding via gene expression and metabolic change via omics approach.

Bacteremia due to Serratia rubidaea in intensive care unit: a case series.

INTRODUCTION: Bacteremia caused by Serratia rubidaea is seldom mentioned in comparison with other Enterobacteriaceae species. It primarily affects immunocompromised patients undergoing invasive procedures. Furthermore, the incidence, clinical features, and microbiological profile of this pathogen in the intensive care unit are rarely described. CASE PRESENTATION: We present four North African case studies of bacteremia in four young female patients admitted to the intensive care unit for ketoacidosis with a history of diabetes mellitus. All four patients developed catheter-related infections complicated by deep vein thrombosis. The catheter site was femoral in all cases, and the main clinical manifestation was poorly tolerated fever. The pathogen was isolated in multiple peripheral blood cultures (> 4) for each patient, showing a similar profile in all cases: resistance to third-generation cephalosporins and sensitivity to aminoglycosides, piperacillin, fluoroquinolones, and folate-pathway inhibitors. Targeted treatment consisted of a combination of ciprofloxacin 400 mg twice per day and trimethoprim/sulfamethoxazole 400/80 mg thrice per day for all four cases. However, in one case, this regimen was switched to amikacin due to adverse effects. The outcomes were favorable in the majority of cases. The patients described in this study were 21, 66, 22, and 27-year-old North African women. CONCLUSION: Most of the reported cases shared common risk factors and clinical aspects. Notably, a case of thrombosis complicating a catheter infection caused by Serratia rubidaea has not been previously reported in the literature. Furthermore, this bloodstream infection typically affects deeply immunocompromised patients. However, our four cases, admitted to the intensive care unit for ketoacidosis, only had a history of diabetes mellitus.

Antagonistic activity of Bacillus amyloliquefaciens subsp. amyloliquefaciens against multidrug resistant Serratia rubidaea.

Serratia rubidaea is an opportunistic Gram-negative pathogen that has developed antimicrobial resistance to a variety of commercial antibiotics. The spread of this multidrug-resistant pattern predicts that it will get harder and harder to treat S. rubidaea infections in the future. For this perception, antimicrobial proteins might represent a safe, effective, and biodegradable alternative because their site of action is on cyclic peptides. In this study, one candidate Bacillus amyloliquefaciens subsp. amyloliquefaciens was isolated from the soil of Sundarban mangrove forest, and its identification was confirmed both using the PCR (Polymerase chain reaction) method and the BIOLOG™ microbial identification system. The antibacterial protein, which has a molecular mass of about 50 kDa, was isolated from B. amyloliquefaciens subsp. amyloliquefaciens. Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was used to confirm the extracted protein's purity. This potential protein was discovered to develop and exhibit antagonistic activity throughout a broad temperature, pH, and salinity range. At doses ranging from 300 to 400 µg/ml, this protein has antagonistic activity against multidrug resistant S. rubidaea and a wide range of other resistant pathogenic bacteria such as Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and so on. The research provides new insights to develop bio-control agents that can be applied for prevent, treat, and control infectious diseases caused by multidrug resistant S. rubidaea, as well as other pathogenic bacteria.

Characterization of the Mechanism of Action of Serratia rubidaea Mar61-01 against Botrytis cinerea in Strawberries.

Several bacterial strains belonging to Serratia spp. possess biocontrol capability, both against phytopathogens and human pathogenic species, thanks to the production of secondary metabolites, including as a red-pink, non-diffusible pigment, 2-methyl-3-pentyl-6-methoxyprodiginine (prodigiosin). Botrytis cinerea is the causal agent of gray mold, which is an economically relevant disease of many crops worldwide. Gray mold is normally controlled by chemical fungicides, but the environmental and health concerns about the overuse of pesticides call for environmentally friendly approaches, such as the use of biocontrol agents. In this study, the efficacy of a specific strain of Serratia rubidaea (Mar61-01) and its metabolite prodigiosin were assessed against B. cinerea under in vitro and in vivo conditions. This strain was effective against B. cinerea, and the effect of prodigiosin was confirmed under in vitro and in vivo conditions. The strain suppressed mycelial growth of B. cinerea (71.72%) in the dual-culture method. The volatile compounds produced by the strain inhibited mycelial growth and conidia germination of B. cinerea by 65.01% and 71.63%, respectively. Efficacy of prodigiosin produced by S. rubidaea Mar61-01 on mycelial biomass of B. cinerea was 94.15% at the highest concentration tested (420 µg/mL). The effect of prodigiosin on plant enzymes associated with induction of resistance was also studied, indicating that the activity of polyphenol oxidase (PPO), superoxide dismutase (SOD) and phenylalanine ammonia lyase (PAL) were increased when prodigiosin was added to the B. cinerea inoculum on strawberry fruits, while catalase (CAT) and peroxidase (POD) did not change. In addition, the volatile organic compounds (VOCs) produced by S. rubidaea Mar61-01 reduced mycelial growth and inhibited conidial germination of B. cinerea in vitro. The findings confirmed the relevant role of prodigiosin produced by S. rubidaea Mar61-01 in the biocontrol of B. cinerea of strawberries, but also indicate that there are multiple mechanisms of action, where the VOCs produced by the bacterium and the plant-defense reaction may contribute to the control of the phytopathogen. Serratia rubidaea Mar61-01 could be a suitable strain, both to enlarge our knowledge about the potential of Serratia as a biocontrol agent of B. cinerea and to develop new biofungicides to protect strawberries in post-harvest biocontrol.

Evidence for enhanced dissipation of chlorpyrifos in an agricultural soil inoculated with Serratia rubidaea strain ABS 10.

The insecticide 14C-chlorpyrifos was found mineralized in a Tunisian soil with repeated exposure to it. From this soil, a bacterial strain was isolated that was able to grow in a minimal salt medium (MSM) supplemented with 25 mg L-1 of chlorpyrifos. It was characterized as Serratia rubidaea strain ABS 10 using morphological and biochemical analyses, as well as 16S rRNA sequencing. In a liquid culture, the S. rubidaea strain ABS 10 was able to dissipate chlorpyrifos almost entirely within 48 h of incubation. Although the S. rubidaea strain ABS 10 was able to grow in an MSM supplemented with chlorpyrifos and dissipate it in a liquid culture, it was not able to mineralize 14C-chlorpyrifos. Therefore, it can be concluded that the dissipation capability of this bacteria might be attributed to its capacity to adsorb CHL. It can also be ascribed to other reasons such as the formation of biogenic non-extractable residues. In both non-sterile and sterile soil inoculated with S. rubidaea strain ABS 10, chlorpyrifos was more rapidly dissipated than in controls with DT50 of 1.38 and 1.05 days, respectively.

Biosurfactant production by Serratia rubidaea SNAU02 isolated from hydrocarbon contaminated soil and its physico-chemical characterization.

The aim of the study was to characterize and optimize the growth media for biosurfactant production from Serratia rubidaea SNAU02 isolated from hydrocarbon-contaminated soil from Cuddalore district, Tamilnadu, India. The biosurfactant produced by S. rubidaea SNAU02, was able to reduce the surface tension to 34.4 mN m(-1) in MSM medium. The biosurfactant was characterized by FT-IR and GC-MS analysis. The GC-MS analysis shows that dirhamnolipid was detected in abundance as predominant congener than monorhamnolipid. The response surface methodology (RSM) -central composite design (CCD) was performed to optimize the media for biosurfactant production. The maximum emulsification index was obtained under the optimal condition of 29.31 g L(-1) mannitol; 2.06 g L(-1) yeast extract, medium pH 6.97 and 5.69 g L(-1) NaCl. The biosurfactant produced by S. rubidaea recovered 92% of used engine oil adsorbed to a sand sample, suggested the potential application in microbial enhanced oil recovery and bioremediation.