Extraction Of Prodigiosin Research Articles

Downstream Process Development for Extraction of Prodigiosin: Statistical Optimization, Kinetics, and Biochemical Characterization

Prodigiosin is natural red colourant derived from Serratia marcescens. However, the high cost of prodigiosin restricts its use in food and pharmaceutical industries, which can be addressed with the design of a suitable extraction procedure. Therefore, the present study aims to use Taguchi methodology to optimize various process parameters during ultrasound-assisted extraction (UAE) to get a higher prodigiosin extraction yield. The most significant contribution comes from the solid-to-liquid ratio (36.66%), followed by sonication of duty cycle (34.82%), medium pH (15.7%), and acoustic intensity (12.82%). The Taguchi technique predicts the highest optimal yield using the solid-liquid ratio (0.3g/mL), duty cycle sonication (75%), acoustic intensity (12.5 w/cm2), and medium pH (3) as parameters. When the extraction conditions were optimized, the yield of prodigiosin increased by 4166.89mg/L. In the future, the above extraction conditions determined using Taguchi approach will be applied for large-scale extraction of prodigiosin. Finally, a second-order kinetic model is used to suit the batch extraction investigation and the second-order rate constant (k) has a value of 4 × 10-5 L/mg/min. In the future, the rate constant, which is reported for the first time, will be used to create a batch extractor for commercial extraction of prodigiosin. Prodigiosin has also been shown to have substantial antioxidant and scavenging properties, which increase in a dose-dependent way with prodigiosin concentration. Because of its antioxidant and scavenging properties, prodigiosin can be used as food additives or pharmaceutical ingredients in industries.

Impact of prodigiosin on staphylococcus aureus isolated from acne vulgaris

Acne is a skin disorder characterized by clogged hair follicles and chronic inflammation of the pilosebaceous follicles. A total of one hundred and five acne swabs were collected from patients with acne vulgaris infection at Baghdad's hospitals. For primary identification, 54 (45%) samples had a positive S. aureus culture on Mannitol salt agar (MSA) for 24 hours at 37°C. In addition, four pre-isolated also identified S. marcescens (pigment producers) which were identified by vitek-2 technique were taken for prodigiosin extraction, mineral salt broth with peptone (0.5 percent) was utilized, followed by pigment extraction, partial purification, also pigment measurement by spectrophotometer. Furthermore, using a microtiter plate assay, the bacterial ability to form biofilms was assessed for 20 S. aureus isolates (multidrug resistant isolates). The results revealed that only 8 strong isolates were biofilm producers, while 5 in addition 7 isolates were moderate also weak biofilm producers. The minimal inhibitory concentration of tetracycline also prodigiosin generated by isolated S. marcescens were determined using the broth microdilution technique for S. aureus isolates (S7 also S8). From overnight cultures of S. aureus (S7 also S8), DNA was effectively extracted.

Impact of prodigiosin on staphylococcus aureus isolated from acne vulgaris

Acne is a skin disorder characterized by clogged hair follicles and chronic inflammation of the pilosebaceous follicles. A total of one hundred and five acne swabs were collected from patients with acne vulgaris infection at Baghdad's hospitals. For primary identification, 54 (45%) samples had a positive S. aureus culture on Mannitol salt agar (MSA) for 24 hours at 37°C. In addition, four pre-isolated also identified S. marcescens (pigment producers) which were identified by vitek-2 technique were taken for prodigiosin extraction, mineral salt broth with peptone (0.5 percent) was utilized, followed by pigment extraction, partial purification, also pigment measurement by spectrophotometer. Furthermore, using a microtiter plate assay, the bacterial ability to form biofilms was assessed for 20 S. aureus isolates (multidrug resistant isolates). The results revealed that only 8 strong isolates were biofilm producers, while 5 in addition 7 isolates were moderate also weak biofilm producers. The minimal inhibitory concentration of tetracycline also prodigiosin generated by isolated S. marcescens were determined using the broth microdilution technique for S. aureus isolates (S7 also S8). From overnight cultures of S. aureus (S7 also S8), DNA was effectively extracted.

Extraction of prodigiosin using aqueous two phase system

Aqueous two-phase system is a modified and amazing separation process of bioproducts in downstream since it keeps their properties. This method is able to give high recovery yield and high purity in a single step. Prodigiosin pigment was produced by Serratia marcescens in carbohydrate fermentation. It is the first time to extract the prodigiosin by aqueous two-phase system, where two types of system are applied; polymer/polymer and polymer/salt. The parameters studied on the recovery per cent and partition coefficient of prodigiosin are temperature, weight fraction of polymer in solution, molecular weight of polymer, weight fraction of salt in solution, and pH. The maximum recovery and partition coefficient of prodigiosin are obtained in the first system (PEG6000/Dextran) at temperature of 20°C, pH 6, weight fraction of PEG6000 is 0.22, weight fraction of Dextran is 0.15 while in the second system (PEG1500/ammonium sulfate) the maximum recovery and partition coefficient of the prodigiosin are obtained at temperature of 20°C, pH 5, weight fraction of salt is 0.15, and weight fraction of PEG1500 is 0.24.

Biocolorant “prodigiosin” interferes with the growth of biofouling bacteria: an in vitro and in silico approach

PurposeThe purpose of this paper was to identify Serratia marcescens to extract and purify prodigiosin pigment to evaluate the antibacterial potential of the pigment.Design/methodology/approachSamples were collected from shrimp aquaculture ponds. Species identification was conducted using morphological, biochemical and molecular tests. Pigment extraction and purification were carried out using column chromatography. The antibacterial effect of crude and purified prodigiosin pigment was evaluated on Escherichia coli, Bacillus subtilis, Pseudomonas aeruginosa and Staphylococcus aureus as biofouling bacteria. In addition, the interaction between prodigiosin and proteins involved in biofilm formation was evaluated using molecular docking.FindingsThe results of prodigiosin extraction with solvents showed the highest percentage of pigment presence with methanol solvent in the second day of culture. The chemical structure of pure prodigiosin obtained from the column chromatography was confirmed by Fourier-transform infrared spectroscopy. Both crude and purified pigments exhibited antibacterial effects against selected bacterial strains. The antibacterial effect of the purified pigment was higher, and the highest antibacterial effect was observed on B. subtilis. Prodigiosin docking was carried out with all target proteins, and the docked energy in all of them was at an acceptable level.Originality/valueProdigiosin extracted from S. marcescens can be used as a bioactive compound to design and manufacture of anti-biofouling and anti-biofilm formation products to use extensively for industrial applications as a natural color in marine industries, food industry, cosmetics and textile productions.

Extraction of prodigiosin from Serratia marcescens and its application as an antibacterial spray

Prodigiosin is a natural pigment produced by Serratia marcescens. The pigment was grown in MacConkey agar and Luria Bertani agar. Growth of the pigment was higher in Luria Bertani agar when compared with MacConkey agar. Antibacterial activity was evaluated using test organisms such as E.coli, Pseudomonas sp. by well diffusion method. The pigment was impregnated on the surface of the test organism in different concentrations and incubated. The clear inhibition zones that formed around the well indicates antibacterial activity. Keywords: Ecoli. Prodigiosin. Pseudomonas sp. Serratia marcescens.

Functioned silver nanoparticle loaded activated carbon for the recovery of bioactive molecule from bacterial fermenter for its bactericidal activity

A novel continuous production and extraction of bacterial bioactive prodigiosin (PG) from fermented using silver nanoparticle impregnated functioned activated carbon composite is proposed for cost-effective and ecofriendly microbial technique. Hence, in this investigation silver nanoparticle was impregnated onto functioned activated carbon ([AC]F) as a support matrix and to enable the separation of PG conjugated silver nanoparticle from the fermented medium. A laboratory scale experiment was carried out to evaluate the continuous production and recovery of PG using [AC@Ag]F. Ag nanoparticle impregnated [AC]F ([AC@Ag]F) characterized by FT-IR, XRD, TGA, DSC and SEM. Instrumental analyses confirmed that Ag nanoparticles significantly impregnated on AC through the functionalization of AC with diethanolamine and it enhances the binding capacity between AC and Ag. The various process parameters, such as contact time, pH, and mass of [AC@Ag]F, were statistically optimized for the recovery of PG using Response Surface Methodology (RSM). The maximum extraction of PG in [AC@Ag]F was found to be 16.2±0.2mgg−1, its twofold higher than [AC]F. Further, PG conjugated [AC@Ag]F and ([AC@Ag]F-PG) were checked for the growth inhibition of gram negative and gram positive bacteria without formation of biofilm upto 96h. Hence, the developed matrix could be eco-friendly, viable and lower energy consumption step for separation of the bacterial bioactive PG from fermented broth. In additionally, [AC@Ag]F-PG was used as an antifouling matrix without formation of biofilm.

Synthesis of Surface-Modified Iron Oxides for the Solvent-Free Recovery of Bacterial Bioactive Compound Prodigiosin and Its Algicidal Activity.

Prodigiosin (PG) is a bioactive compound produced by several bacterial species. Currently, many technologies are being developed for the production of PG by fermentation processes. However, new challenges are being faced with regard to the production of PG in terms of the recovery and purification steps, owing to the labile nature of PG molecules and the cost of the purification steps. Conventional methods have limitations due to high cost, low reusability, and health hazards. Hence, the present investigation was focused on the development of surface-functionalized magnetic iron oxide ([Fe3O4]F) for solvent-free extraction of bioactive PG from the bacterial fermented medium. Fe3O4 was functionalized with diethanolamine and characterized by FT-IR, diffuse reflectance spectroscopy, thermogravimetric analysis, scanning electron microscopy, and confocal microscopy. The various process parameters, such as contact time, temperature, pH, and mass of Fe3O4, were optimized for the extraction of PG using functionalized Fe3O4. Instrumental analyses confirmed that the PG molecules were cross-linked with functional groups on [Fe3O4]F through van der Waals forces of attraction. PG extracted through Fe3O4 or [Fe3O4]F was separated from the fermentation medium by applying an external electromagnetic field and regenerated for successive reuse cycles. The purity of the extracted PG was characterized by high-performance liquid chromatography, FT-IR, and UV-visible spectroscopy. The iron oxide-diethanolamine-PG cross-linked ([Fe3O4]F-PG) composite matrix effectively deactivates harmful fouling by cyanobacterial growth in water-treatment plants. The present investigation provides the possibility of solvent-free extraction of bacterial bioactive PG from a fermented medium using functionalized magnetic iron oxide.

A novel method for the extraction of prodigiosin from bacterial fermenter integrated with sequential batch extraction reactor using magnetic iron oxide

A novel method for the extraction of prodigiosin from bacterial fermentation was achieved using an integrated sequential batch extraction reactor (SBER) containing functionalized iron oxides ([Fe3O4]F). The [Fe3O4]F was prepared using diethanolamine to impart quaternary amine group onto Fe3O4. The quaternary amine group imparted to Fe3O4 enhanced the prodigiosin adsorption efficiency. Two SBER were integrated with the continuous fermenter for the continuous extraction of prodigiosin. The present investigation established a method to conserve solvent by about 95% during the prodigiosin extraction step. The proposed extraction method can be regarded as solvent free direct extraction of bacterial bioactive compounds from bacterial fermented medium.

Efficient recombinant production of prodigiosin in Pseudomonas putida.

Serratia marcescens and several other bacteria produce the red-colored pigment prodigiosin which possesses bioactivities as an antimicrobial, anticancer, and immunosuppressive agent. Therefore, there is a great interest to produce this natural compound. Efforts aiming at its biotechnological production have so far largely focused on the original producer and opportunistic human pathogen S. marcescens. Here, we demonstrate efficient prodigiosin production in the heterologous host Pseudomonas putida. Random chromosomal integration of the 21 kb prodigiosin biosynthesis gene cluster of S. marcescens in P. putida KT2440 was employed to construct constitutive prodigiosin production strains. Standard cultivation parameters were optimized such that titers of 94 mg/L culture were obtained upon growth of P. putida at 20°C using rich medium under high aeration conditions. Subsequently, a novel, fast and effective protocol for prodigiosin extraction and purification was established enabling the straightforward isolation of prodigiosin from P. putida growth medium. In summary, we describe here a highly efficient method for the heterologous biosynthetic production of prodigiosin which may serve as a basis to produce large amounts of this bioactive natural compound and may provide a platform for further in-depth studies of prodiginine biosynthesis.

Optimizing Ultrasound-Assisted Extraction of Prodigiosin by Response Surface Methodology

Prodigiosin extraction from dried Serratia marcescens jx1 cells using ultrasound-assisted extraction was optimized. The experiment was carried out in accordance with a central composite design (CCD) three-level and single-variable approach. The extraction time, extraction temperature, and solute to solvent ratio with the application of ultrasonication were optimized using response surface methodology (RSM) to maximize the extraction of prodigiosin from dried S. marcescens jx1 cells. The response of prodigiosin was determined using spectrophotometry. A quadratic model was established to predict the prodigiosin extraction yield. The analysis of variance showed that the quadratic model significantly contributed to the response of prodigiosin. The optimal extraction parameters were an extraction time of 17.5 min, an extraction temperature of 23.4°C, and a solvent-to-solute ratio of 1:27.2. Under these optimum conditions, the average prodigiosin yield was 4.3 g ± 0.02 g from 100 g of dried cells, which matches the predicted values. The obtained optimum conditions for prodigiosin extraction provide a scientific basis for the economical large-scale production of prodigiosin.

Physical and chemical stress on Serratia marcescens and studies on prodigiosin pigment production

Bio-pigments play a very important role in many industries such as food, feed, cosmetics and pharmaceutical products. The red pigment prodigiosin produced by Serratia marcescens possesses a lot of medical benefits. The employment of ultraviolet (UV) radiation and ethyl methanesulfonate (EMS) to induce variants with the enhanced pigmentation of S. marcescens is the main scope of this study. Also, the antimicrobial activity and the cytotoxicity properties of prodigiosin were investigated. Subsequent to physical and chemical stress induced on S. marcescens, many variants are selected. They showed different patterns in growth at 38 °C, salinity tolerance, emulsification and hemolytic activities compared to the parent S. marcescens. The EMS-variant (S26) produced prodigiosin eight-fold higher than that of the parent strain. Three new media are described for S. marcescens’s growth and pigmentation. They are based on the market-refused squash seeds; sunflower seeds and watermelon seeds. The present study modifies and designs a new procedure for prodigiosin extraction using small volumes of bacterial cultures. Prodigiosin was purified using column chromatography; it was found that prodigiosin is susceptible to non-polar solvents. Crude prodigiosin extracted from variant (S26) is stable at alkaline pH and its thermal stability is up to 80 °C. The experiment of testing the dyeing properties of prodigiosin is outlined and its results are deeply encouraging for the use of this interesting pigment for ultimate medical applications.

A novel medium for the enhanced cell growth and production of prodigiosin from Serratia marcescens isolated from soil

BackgroundProdigiosin produced by Serratia marcescens is a promising drug owing to its reported characteristics of having antifungal, immunosuppressive and antiproliferative activity. From an industrial point of view the necessity to obtain a suitable medium to simultaneously enhance the growth of Serratia marcescens and the pigment production was the aim of this work. The usage of individual fatty acid as substrate in industries would be cost-effective in the long run and this paved the way for us to try the effect of different fatty acid-containing seeds and oils of peanut, sesame and coconut as source of substrate.ResultsThe addition of sugars only showed slight enhancement of prodigiosin production in nutrient broth but not in fatty acid containing seed medium. The powdered peanut broth had supported better growth of Serratia marcescens and higher yield of prodigiosin when compared with the existing nutrient broth and peptone glycerol broth. A block in prodigiosin production was seen above 30°C in nutrient broth, but the fatty acid seed medium used by us supported prodigiosin production upto 42°C though the yields were lower than what was obtained at 28°C. From the results, the fatty acid form of carbon source has a role to play in enhanced cell growth and prodigiosin production.ConclusionWe conclude by reporting that the powdered and sieved peanut seed of different quality grades were consistent in yielding a fourty fold increase in prodigiosin production over the existing media. A literature survey on the composition of the different media components in nutrient broth, peptone glycerol broth and the fatty acid containing seeds and oils enabled us to propose that the saturated form of fatty acid has a role to play in enhanced cell growth and prodigiosin production. This work has also enabled us to report that the temperature related block of prodigiosin biosynthesis varies with different media and the powdered peanut broth supports prodigiosin production at higher temperatures. The medium suggested in this work is best suitable from an industrial point of view in being economically feasible, in terms of the higher prodigiosin yield and the extraction of prodigiosin described in this paper is simple with minimal wastage.

Extraction of Prodigiosin from Blood Plasma and Body Tissues and Its Measurement by Spectrophotometric Analysis

Satisfactory methods have been developed to extract prodigiosin from samples of blood plasma and various body tissues. They depend on the preferential solubility of the dye in petroleum ether or in acid chloroform over that in blood plasma or in tissue fluids. The concentration of prodigiosin in these extracts may be measured accurately by spectrophotometric analysis, using the appropriate wave lengths of the absorption band of the dye in the solvents Amounts as small as 0.1 micrograms per milliliter may be detected in a reproducible manner.