Red Pigment Production Research Articles

Production and extraction of red pigment by solid-state fermentation of broken rice using Monascus sanguineus NFCCI 2453

The present work has attempted to use broken rice as an economical substrate for the solid-state fermentation (SSF) mediated by Monascus sanguineus NFCCI 2453 for red pigment production. UV–Vis spectral and FTIR analyses were employed for the confirmation of the red pigment production. Response surface methodology (RSM) was used to optimize four significant parameters (extraction time, ethanol concentration, liquid-to-solid ratio and agitation speed) for extracting red pigment from the fermented biomass. Results from ANOVA proved the developed model to be significant as the R2 value (0.9602) demonstrated that the experimental values matched well with the results, as predicted. The maximum red pigment yield of 143.3 ODU/gds was obtained with the optimal setting of the process parameters. The model was found to be accurate (99.3%) using validation experimental data. The study serves as a prelude for exploiting M. sanguineus for the efficient production of red pigment using a low-cost substrate.

Agro-industrial residues for the production of red biopigment by Monascus ruber: rice flour and sugarcane molasses.

Three culture media were studied for red pigment production by Monascus ruber in submerged cultivation: rice flour (20 g L-1), sugarcane molasses (30 g L-1), and, finally, molasses + rice flour (10 g L-1+10 g L-1); all culture media were added of 5 g L-1 glycine as nitrogen source. Rice flour showed pigment production of 7.05 UA510nm and molasses 5.08 UA510nm, and the mixture of rice flour and molasses showed the best result of 16.38 UA510nm. Molasses culture presented good results for cell biomass production of 11.09 g L-1. With these results, it was observed that one substrate presented good pigment production (rice flour) and another attained better results for cell biomass growth (molasses), and a third medium containing 10 g L-1 of rice flour + 10 g L-1 of molasses was formulated. The results for this mixture showed satisfactory results, with global pigment productivity of 0.097 UA510nm h-1 and maximum productivity rate of 0.17 UA510nm h-1. The high production and productivity obtained for the mixture of rice flour and molasses indicated that the production of red pigment by submerged fermentation, using the mixture of these low-cost culture media, may be promising in terms of commercial production.

IMPROVEMENT OF ORGANIC RED PIGMENT PRODCUTION BY MONASCUS PURPUREUS TISTR3651 USING PATHUMTHANI-1 RICE-BASED MEDIUM IN SUBMERGED AND SOLID-STATE FERMENTATION

Objective: This research is to study the production of natural red pigment by Monascus purpureus TISTR3615 in the submerged and solid-statefermentation system using Pathumthani-1 rice as a carbon source.Methods: The antioxidant activity of the red pigment was evaluated in vitro 2,2-diphenyl-1-picrylhydrazyl (DPPH), ABTS radical scavenging assay,and ferric-reducing antioxidant power (FRAP) assay, including total phenolic compound.Results and Discussion: The maximum of red pigment production was 0.55±0.02/ml (OD 680 nm) after incubation at 30°C for 24 days. Theantioxidant activity based on inhibition DPPH (%), ABTS radical scavenging activity (%), and FRAP activity (mM Fe2+/g) was 97.80±1.51,68.64±0.46, and 0.32±0.021, respectively. The total phenolic content was 164.78±2.82 μg GAE/mg.Conclusion: It was estimated that Monascus pigments, leading to nutraceutical and pharmaceutical applications, cosmetic industry, and foodindustry.

Study on Extraction Technology and Stability of Red Pigment in <i>Red Plumeria</i>

Background: Pigments, especially for those red color are widely used in various products and are closely related to human health, however, the study on screening optimal conditions for extracting red pigment from red frangipani is less reported. Objective: This study is aimed to decipher the effects of these factors on the stability of the pigment. Method: We developed an optimized extraction method of red pigment from Plumeria rubra L leaves through different combinations of extractants, incubation time and temperature. Results: Results show that the largest productivity of red pigment is found under the condition of 10% citric acid as extractant at 80°C for 90 min compared to other conditions. The pigment is sensitive to high light and appears to have strong reducibility rather than oxidability. The pigments production is also sensitive to pH value, as well as metal ion strength, such as Al3+, Cu2+, Fe3+, Mn2+, Zn2+, K+, Mg2+, Na+, and Ca2+, whereas the production is inhibited by adding sucrose, salt and soluble starch. Conclusion: We concluded that red pigment is sensitive to external environmental stimulus and internal ion and carbohydrates concentrations, among these factors, a combination of 10% citric acid extranctant together with 80oC for 90 min treatments ensures to obtain the largest productivity of red pigment. This study provides a fundamental basis for the production of natural red plumeria red pigment and the screening for edible pigment varieties.

Antimicrobial and Antioxidant potentials of Carotenoid Pigment Produced by Indigenous Novel Soil Isolate Rhodococcus kroppenstedtii

The present research study evaluated the antioxidant and antimicrobial potentials of intense red carotenoid pigment produced by novel soil isolate.  A novel isolate from the rhizospheric soil of the soya-bean field was found to produce prominent red carotenoid pigment. Based on  phenotype, biochemical and molecular features (16S rRNA sequencing), the isolate was identified as Rhodococcus kroppenstedtii.  Maximum intense red pigment production appeared in yeast extract ma

Enhancement of microbial pigment production from Monascus ruber by sodium octanoate addition.

The addition of fatty acids and other molecules to culture media may intensify the production of biomolecules, such as monascus pigments, however, few studies of this have been developed. Thus, the objective of the present study was to investigate the effects of adding sodium octanoate to the culture medium, with a view to increasing the synthesis and production of the pigments produced by Monascus ruber CCT 3802 on solid and submerged cultivations. Monacus ruber CCT 3802 was cultivated on solid and submerged media supplemented with different concentrations of sodium octanoate. The radial growth rate of the colonies was obtained from the declivity of the linear regression of the radius of the colonies as a function of cultivation time and the kinetics of submerged cultivations were performed. The filtrate obtained was submitted to scanning spectrophotometry at a range from 350 to 550 nm and the color parameters were determined by using the CIELAB color system. The data were submitted to a univariate analysis of variance (ANOVA) and the means obtained for each treatment submitted to Tukey's test using Statistica version 5.0 software at a 5% level of significance. Sodium octanoate exerted a strong influence on growth and pigment production in solid and submerged cultivations. The values for L*, a* and b* were positive for pigments produced, with regards to colors close to red and yellow. In the media supplemented with 1.0 mM and 1.5 mM of sodium octanoate, the production of red pigments became expressive from 48 hours-cultivation, increasing considerably from the second to the fourth days. This shows that supplementation with sodium octanoate provides a greater production of pigments in a shorter time interval than the control culture, which required 144 hours of cultivation to present a higher value for AU510nm, which directly influenced pigment productivity. The addition of sodium octanoate exerted a significant influence on both microbial growth and pigment production in both solid and submerged cultivations. The supplementation of the submerged cultures with sodium octanoate was responsible for an expressive production of pigments in just 48 hours, whereas 144 hours were necessary in the absence of sodium octanoate. These results are promising for increasing the productivity of pigment production, including possibilities for application on an industrial scale.

Enhancement of microbial pigment production from Monascus ruber by sodium octanoate addition [pdf

BACKGROUND The addition of fatty acids and other molecules to culture media may intensify the production of biomolecules, such as monascus pigments, however, few studies of this have been developed. Thus, the objective of the present study was to investigate the effects of adding sodium octanoate to the culture medium, with a view to increasing the synthesis and production of the pigments produced by Monascus ruber CCT 3802 on solid and submerged cultivations. METHODS Monacus ruber CCT 3802 was cultivated on solid and submerged media supplemented with different concentrations of sodium octanoate. The radial growth rate of the colonies was obtained from the declivity of the linear regression of the radius of the colonies as a function of cultivation time and the kinetics of submerged cultivations were performed. The filtrate obtained was submitted to scanning spectrophotometry at a range from 350 to 550 nm and the color parameters were determined by using the CIELAB color system. The data were submitted to a univariate analysis of variance (ANOVA) and the means obtained for each treatment submitted to Tukey's test using Statistica version 5.0 software at a 5% level of significance. RESULTS Sodium octanoate exerted a strong influence on growth and pigment production in solid and submerged cultivations. The values for L*, a* and b* were positive for pigments produced, with regards to colors close to red and yellow. In the media supplemented with 1.0 mM and 1.5 mM of sodium octanoate, the production of red pigments became expressive from 48 hours-cultivation, increasing considerably from the second to the fourth days. This shows that supplementation with sodium octanoate provides a greater production of pigments in a shorter time interval than the control culture, which required 144 hours of cultivation to present a higher value for AU510nm, which directly influenced pigment productivity. CONCLUSIONS The addition of sodium octanoate exerted a significant influence on both microbial growth and pigment production in both solid and submerged cultivations. The supplementation of the submerged cultures with sodium octanoate was responsible for an expressive production of pigments in just 48 hours, whereas 144 hours were necessary in the absence of sodium octanoate. These results are promising for increasing the productivity of pigment production, including possibilities for application on an industrial scale.

Isolation and Optimization of Monascus ruber OMNRC45 for Red Pigment Production and Evaluation of the Pigment as a Food Colorant

The color of food is a critical factor influencing its general acceptance. Owing to the effects of chemical colorants on health, current research is directly aimed at producing natural and healthy food colorants from microbial sources. A pigment-producing fungal isolate, obtained from soil samples and selected based on its rapidity and efficiency in producing red pigments, was identified as Monascus ruber OMNRC45. The culture conditions were optimized to enhance pigment production under submerged fermentation. The optimal temperature and pH for the highest red pigment yield were 30 °C and 6.5, respectively. The optimum carbon and nitrogen sources were rice and peptone, respectively. The usefulness of the pigment produced as a food colorant was evaluated by testing for contamination by the harmful mycotoxin citrinin and assessing its biosafety in mice. In addition, sensory evaluation tests were performed to evaluate the overall acceptance of the pigment as a food colorant. The results showed that M. ruber OMNRC45 was able to rapidly and effectively produce dense natural red pigment under the conditions of submerged fermentation without citrinin production. The findings of the sensory and biosafety assessments indicated the biosafety and applicability of the red Monascus pigment as a food colorant.

Alternative Extraction and Characterization of Nitrogen-Containing Azaphilone Red Pigments and Ergosterol Derivatives from the Marine-Derived Fungal Talaromyces sp. 30570 Strain with Industrial Relevance.

Many species of Talaromyces of marine origin could be considered as non-toxigenic fungal cell factory. Some strains could produce water-soluble active biopigments in submerged cultures. These fungal pigments are of interest due to their applications in the design of new pharmaceutical products. In this study, the azaphilone red pigments and ergosterol derivatives produced by a wild type of Talaromyces sp. 30570 (CBS 206.89 B) marine-derived fungal strain with industrial relevance were described. The strain was isolated from the coral reef of the Réunion island. An alternative extraction of the fungal pigments using high pressure with eco-friendly solvents was studied. Twelve different red pigments were detected, including two pigmented ergosterol derivatives. Nine metabolites were identified using HPLC-PDA-ESI/MS as Monascus-like azaphilone pigments. In particular, derivatives of nitrogen-containing azaphilone red pigment, like PP-R, 6-[(Z)-2-Carboxyvinyl]-N-GABA-PP-V, N-threonine-monascorubramin, N-glutaryl-rubropunctamin, monascorubramin, and presumed N-threonyl-rubropunctamin (or acid form of the pigment PP-R) were the major pigmented compounds produced. Interestingly, the bioproduction of these red pigments occurred only when complex organic nitrogen sources were present in the culture medium. These findings are important for the field of the selective production of Monascus-like azaphilone red pigments for the industries.

Direct conversion of waste loquat kernels to pigments using Monascus purpureus ATCC16365 with proteolytic and amylolytic activity

Kernels of loquat (Eriobotrya japonica L.) fruits are rich in protein and starch contents; however, they have no significant application in world. This study was performed to produce pigments from Monascus purpureus ATCC16365 using loquat kernel powder (LKP) as a fermentation substrate and investigate the effect of four mineral salts (CaCl2, KH2PO4, MgSO4, and FeSO4) on pigment production in LKP-based medium. During the experiments, LKP was not subjected to any chemical or enzymatic hydrolysis, and it was tested directly as a substrate in shaking flask and fermenter cultures. It was found that addition of CaCl2 alone decreased red pigment production but did not cause an important change in synthesis of yellow and orange pigments. Supplementation of KH2PO4, MgSO4, or FeSO4 alone decreased the production of yellow pigment but increased the production of red pigments. When three salts (1.0 g/L KH2PO4, 0.3 g/L MgSO4, and 0.03 g/L FeSO4) were added together to the LKP-based medium, more productions of enzymes (protease and amylase) and red pigments were achieved. When experiments were performed in optimized medium (LKP and three mineral salts), maximum concentrations of red, orange, and yellow pigments were determined as 292, 193, and 171 AU/L in flask culture but 327, 241, and 204 AU/L in fermenter culture, respectively. Amylase and protease activities were 93.4 and 52.3 U/L in flask culture but 93.8 and 52.8 U/L in fermenter culture, respectively. This is the first report on the use of LKP as a substrate in production of enzymes and pigments from M. purpureus.

Factors Affecting Red Pigment Production by Local Fungal Isolate of Penicillium sp.

Factors Affecting Red Pigment Production by Local Fungal Isolate of Penicillium sp.

Safe natural bio-pigment production by Monascus purpureus using mixed carbon sources with cytotoxicity evaluation on root tips of Allium cepa L.

By increasing the undesirable side effects of synthetic food pigments on human health, using safe natural food pigment become an urgent issue. Incorporate corn starch with oils conducted a high impact on red pigment production by Monascus purpureus. Fortification the medium with sesame oil raised the pigment production by 80% and the dry mass by 63% compared with free oil medium. Response surface methodology maximizes the production with 114.6% (12.8 A500) using medium constituents (g/l); Sesame oil 5; Corn starch 30; Yeast extract 1.5; KH2PO4 2.5 and MgSO4.7H2O 0.1. After evaluating red pigment stability in three common food components, citric acid showed a great effect on residual stability percentage compared with ascorbic and salicylic acid which decrease slightly the residual stability percentage at light and dark conditions. The mitotic index of red pigment was lower than the negative control at all tested concentrations. Different types of mitotic chromosomal abnormalities e.g. lagging chromosome, chromosomal bridge, chromosome and chromatin fragments, outside chromosome, chromosomal stickiness and micro nuclei were recorded. Insignificant increase in total mitotic aberrations percentage in all tested root tips treated with all concentrations of red pigment (1.23, 1.58, 1.63, 2.32 and 2.40%) compared with negative control (0.91%). There was a significant increase in total aberrations percentage after treatment with all concentrations (10, 15, 20 and 25%) of positive control (2.93, 3.00, 3.55 and 6.53 respectively) except (5%) which was insignificant (2.71%). From the previous data, this red pigment can be used as an alternative safe pigment in the food industry.

Comparison of Two Methods for Counting Molds in Fermentations Using the Production of Bikaverin by Fusarium oxysporum CCT7620 as a Model.

The dry cell weight (DCW) measurement is one of the preferred methods to determine the growth of filamentous fungi. However, this technique is not applicable to insoluble culture media, besides being possibly influenced by the presence of extracellular biomass. The standard plate counting (SPC) is a reference method for detecting viable cells; however, it is referred as imprecise. In this study, we did a comprehensive analysis of the errors associated to each procedure and also determined the growth kinetics of Fusarium oxysporum in soluble (DCW and SPC) and insoluble (SPC) culture media. Finally, we used the production of bikaverin in airlift bioreactor containing insoluble medium as a case study to estimate red pigment production and to monitor biomass growth via SPC. We concluded that SPC can be used to give reliable fungal growth kinetics in media with insoluble matter, yielding errors equivalent to DCW depending on the number of replicates done for serial dilutions and plate counting.

Pigment production by Fusarium solani BRM054066 and determination of antioxidant and anti-inflammatory properties

The fungal kingdom has been widely studied as a source of bioactive compounds of interest to the pharmaceutical and food industry. This paper studies the production of natural red pigments by Fusarium solani BRM054066 in the submerged fermentation system, using Doehlert experimental design to determine optimal cultivation conditions. The chemical composition of the red pigment was determined by Nuclear Magnetic Resonance spectroscopy (NMR) and Ultra-Performance Liquid Chromatography coupled to Mass Spectrometry (UPLC-MS). Antioxidant activity was assessed by the ability to sequester of free radical DPPH. In the analysis of anti-inflammatory activity, murine peritoneal macrophages activated by LPS were used, and the gene expression of TNF-α, IL-1β, IL-6, IL-10 and IL-17 was determined using qPCR. As a result, it was found that agitation at 200 rpm and glucose concentration ≥ 20 g/L promote the best results in the production of red pigment. The chemical compounds identified were two naphthoquinones, fusarubin and dihydrofusarubin, and an anthraquinone, a bostrycoidin, being fusarubin the majority compound. The red pigment showed antioxidant activity by scavenge 50% of the DPPH radical, in a concentration of 24 µg/mL. The pigment also showed an effective anti-inflammatory capacity by reducing the overexpression of the pro-inflammatory cytokines TNF-α, IL-1β and IL-6 and promoting the production of anti-inflammatory IL-10 and IL-17, in murine macrophages activated by LPS (p < 0.05). According to the results, the fungus F. solani BRM054066, under optimized conditions of cultivation, proved to be a promising source of biologically active natural pigments with wide industrial applicability.

Isolation and identification of two Serratia marcescens strains from silkworm, Bombyx mori.

Bacterial septicemia commonly occurs and usually cause huge losses in sericulture industry. Here, two pathogenic bacterial strains were isolated from dead silkworm and named as ZJ-1 and ZJ-2. Phenotypic and genotypic analysis results revealed that both of these two strains are closely related to Serratia marcescens (S. marcescens). The morphological as well as physiological and biochemical characteristics of ZJ-1 were accordant with S. marcescens mentioned in Bergey's manual of determinative bacteriology, whereas ZJ-2 showed some discrepancies such as the utilization of malonate and starch, fermentation of maltose and sucrose, and tests of urease, etc. Surprisingly, ZJ-2 could produce red pigment at high temperature (37°) but ZJ-1 could not. Besides, by analyzing the lethal concentration 50 (LC50) of ZJ-1 and ZJ-2, it was found that the virulence of ZJ-2 was lower than that of ZJ-1. These results revealed that ZJ-1 and ZJ-2 were two different strains of S. marcescens and that ZJ-2 was expected to be a safe (low-toxicity) and efficient strain for the production of red pigment. Nonetheless, further research in molecular level is needed to understand the regulation mechanism of pigment production and infection of ZJ-2.

A Dual-Plasmid CRISPR/Cas System for Mycotoxin Elimination in Polykaryotic Industrial Fungi.

Mycotoxin contamination causes disease and death in both humans and animals. Monascus Red, produced by Monascus purpureus, is used as a food colorant. However, its application is limited by contamination of the nephrotoxin citrinin, which is also produced by the fungus. Suppressing citrinin production by genetic engineering is difficult in a polykaryotic fungus such as M.purpureus. Hence, we developed a CRISPR/Cas system to delete large genomic fragments in polykaryotic fungi. Protoplast preparation and regeneration were optimized, and a dual-plasmid CRISPR/Cas system was designed to enable the deletion of the 15-kb citrinin biosynthetic gene cluster in M.purpureus industrial strain KL-001. The obtained homokaryotic mutants were stable, and citrinin was unambiguously eliminated. Moreover, the Monascus Red pigment production was increased by 2-5%. Our approach provides a powerful solution to solve this long-standing problem in the food industry and enables engineering of polykaryotic fungi for mycotoxin eliminations.

Evaluation of tyrosol and farnesol as inducer in pigment production by Monascus purpureus ATCC16365.

This study focused on investigating the effect of exogenously applied two quorum sensing molecules (tyrosol and farnesol) on the synthesis of bioactive metabolites (pigments, lactic acid, ethanol, and citric acid) in Monascus purpureus ATCC16365. None of the tested concentrations (62.5, 125, 250, and 500 µl/L) of farnesol affected the synthesis of metabolites as well as cell growth. As with farnesol application, none of the tested concentrations (3.45, 6.9, 13.8, and 27.6 mg/L) of tyrosol caused a significant change in the synthesis of lactic acid and citric acid as well as cell growth. Conversely, all of the tested concentrations of tyrosol increased pigment synthesis but reduced ethanol synthesis, compared with the control. Maximum increases (3.16-, 2.68-, and 2.87-fold increase, respectively) in yellow, orange, and red pigment production were achieved, especially when 6.9-mg/L tyrosol was added to the culture on day 3. On the contrary, 6.9-mg/L tyrosol reduced the content of citrinin by approximately 51.5%. This is the first report on the effect of tyrosol and farnesol on the synthesis of Monascus metabolites. Due to potential properties, such as low price, nonhuman toxicity, promotion of pigment synthesis, and reduction in citrinin synthesis, tyrosol can be used as a novel inducer in the fermentative production of Monascus pigments.

Physicochemical Analysis of Distribution of Useful Components in Waste in the Thermal Energy Sector

The physicochemical analysis of distribution of useful components in processing of ash and slag of the thermal energy sector plants is performed. The article describes thermodynamics and kinetics of chemical reactions during agglomeration of feedstock and ammonium hydrofluoride at the temperatures of 50–200 °C, sublimation of ammonium hexafluorosilicate in the temperature range of 350–550 °C, production of amorphous silica nanoparticles, alumina particles and red iron oxide pigment, as well as formation of calcium fluoride (Ca, Y)F2 which is a concentrator of rare and other elements. The efficient technology is developed for processing of electromagnetic fraction of ash and slag with integrated recovery of various useful components.

Antioxidant Activity and Role of Culture Condition in the Optimization of Red Pigment Production by Talaromyces purpureogenus KKP Through Response Surface Methodology.

The red pigment production by Talaromyces purpureogenus KKP, a soil isolate, was optimized by response surface methodology (RSM) in the present study. The cultural parameters, such as pH, temperature, dextrose, and peptone concentrations, were optimized for red pigment production using the central composite design (CCD) experimental design. A second-order quadratic model was used to calculate the relationships between the values at different levels of response. The optimum values of the selected variables under coded factors are 6.0, 27 °C, 2.25%, and 1.10% for pH, temperature, dextrose, and peptone, respectively. The selected variables were most effective in the enhancement of red pigment production at optimized culture conditions. In addition to optimization, the antioxidant activity of the pigment isolated in the present study was found to be promising with IC50 value (40µg/ml). The HRMS data revealed the identification of delphinidin, limonene, 6-hydroxymethyl-7,8-dihydropterin, D-mannose 6-phosphate, and CDP-DG (18:0/18:0). The results of the present investigation will be added to the existing literature of red pigment production and its optimization by T. purpureogenus.

Enrichment and purification of red pigments from defective mulberry fruits using biotransformation in a liquid-liquid-solid three-phase system.

A large number of defective mulberries are discarded each year because mulberries are easy to break. The red pigments from defective mulberries are recognized as the sustainable sources of anthocyanins extracted from nature. Cyanidin-3-O-rutinoside and cyanidin-3-O-glucoside are the main components of mulberry red pigments, accounting for 50% and 40% of the total, respectively. Cyanidin-3-O-glucoside exhibits anticancer, hypoglycemic, and liver and visceral protection properties. Cyanidin-3-O-glucoside can be prepared by enzymatically hydrolyzing the rhamnosidase bond of cyanidin-3-O-rutinoside. To obtain mulberry red pigment with a high purity of cyanidin-3-O-glucoside, immobilized α-L-rhamnosidase was added to the aqueous two-phase system to construct a liquid-liquid-solid three-phase enzyme catalytic system. After optimization, the three-phase system was composed of 27.12% (w/w) ethanol, 18.10% (w/w) ammonium sulfate, 15% (w/w) mulberry juice, 4.24% (w/w) immobilized α-L-rhamnosidase, and 35.54% (w/w) pure water. The three-phase system was employed to enrich and purify cyanidin-3-O-glucoside at pH 5 and 45°C for 1h. The purity of cyanidin-3-O-glucoside was increased from 40 to 82.42% with cyanidin-3-O-rutinoside conversion of 60.68%. The immobilized α-L-rhamnosidase could be reused seven times, maintaining a relative activity of over 50%. Overall, the developed system provided an efficient and simple approach for high purity mulberry red pigment production and recycling in the field of sustainable agriculture. Graphical abstract.