Bioactive Pigments Research Articles

High-Performance Extraction Process of Anthocyanins from Jussara (Euterpe edulis) Using Deep Eutectic Solvents

New strategies for obtaining target bioactive compounds and natural pigments with the use of “green solvents” are consistently being developed, and deep eutectic solvents are (DES) a great alternative. This work established the significant variables and models for anthocyanin extraction, using DES and experimental design, of Euterpe edulis Mart. (jussara) fruit pulp, an endangered palm tree from the Brazilian Atlantic Forest. From a screening of seven initially tested DES, choline chloride/xylitol-based solvents had the best results with up to 42% increase in the total anthocyanin yield compared to methanolic extraction. Antioxidant assays also revealed a maximum antioxidant capacity of 198.93 mmol Trolox/100 g dry weight basis. The DES extract showed slower degradation to heat at 60° and 90 °C (2.5 times) and indoor constant light source (1.9 times) than methanolic extracts. The optimal extract also revealed slight inhibition of S. enterica and S. aureus growth in the agar plate.

Multi-target bioactivity of summer quinones production in the Persian Gulf burrowing black-type sea urchin

After harvesting the sea urchin gonads for Japanese food “uni” echinoculture systems, the remaining shells and spines are considered waste. However, the material of shells and spines is thought to be rich in natural bioactive molecules. The current study used liquid chromatography–electrospray mass spectrometry to extract summer quinones pigment present in spines and shells of the burrowing sea urchin ‘black’ type Echinometra mathaei from the natural Qeshm Island echinoculture. Then, the biochemical, antioxidant, anti-inflammatory, antidiabetic, antibacterial, and cytotoxic activities of sea urchin quinones pigment were investigated. In terms of bioactivity, both shell and spine pigments demonstrated strong radical scavenging activity (antioxidant). The shell pigment exhibited maximum albumin denaturation inhibition (IC50 = 9.62 μg/ml) (anti-inflammatory), as well as α-amylase inhibition (92.28 percent 4.77) (antidiabetic). Pigments were discovered to have a low antibacterial effect against positive gramme bacteria, as well as low cytotoxic and embryotoxic effects when compared to Artemia salina and zebrafish (Danio rerio). For identification and quantification of pigment extracts, both the photodiode array detector and LC-ESI-MS were used. Spinochrome A, B, and C, as well as echinochrome A, were identified as bioactive quinonoid pigments. This chemical defence is discussed in relation to its algal diet and environmental conditions. In conclusion, the isolated pigments obtained from the shell and spines of E. mathaei sea urchins found to have potent bio-activity and can be used for various biomedical and pharmaceutical applications.

Stress Dependent Biofilm Formation and Bioactive Melanin Pigment Production by a Thermophilic Bacillus Species from Chilean Hot Spring.

Thermophilic bacteria able to survive extreme temperature stress are of great biotechnological interest due to their extracellular production of bioactive molecules as a part of a survival strategy, or by intracellular modifications. In the present study, thermophilic Bacillus haynesii CamB6, isolated from a Chilean hot spring, was studied for the formation of different stress response molecules. The polymeric pigment produced by the bacterial strain was characterized by different physicochemical techniques. On exposure to ranges of temperature (50–60 °C), pH (5.0–7.0), and sources of nitrogen and carbon (1–5 g·L−1), the bacteria responded with a biofilm network formation in a hydrophobic polystyrene surface. Biofilm formation under fed-batch conditions was also statistically validated. The bacteria showed a planktonic pellicle network formation in the presence of induced hypoxia and salinity stress (19.45 g·L−1) under static conditions. Salinity stress also resulted in the intracellular response of brown pigment production. The pigment was structurally and functionally characterized by UV-Vis absorbance and the presence of different characteristic peaks via FTIR analysis (bacterial pyomelanin fingerprints) were assessed. A high thermal stability and TGA profile indicated the brown pigment was a probable pyomelanin candidate. Micropyrolysis (Py-GC/MS) showed that isoprene, pyrrole, benzene, pyridine, and their derivatives were the major components detected. In addition, acetic acid, indole, phenol, and its derivatives were observed. The absence of sulfocompounds in the pyrolyzed products agreed with those reported in the literature for pyomelanin. The pigment surface morphology was analyzed via SEM, and the elemental composition via EDS also demonstrated the similarity of the brown pigment to that of the melanin family. The pyomelanin pigment was observed to be bioactive with promising antioxidant capacity (H2O2, Fe2+) compared to the standard antioxidant molecules. In conclusion, B. haynesii CamB6 demonstrated the formation of several biomolecules as a stress response mechanism that is bioactive, showing its probable biotechnological applications in future.

Biomedical Applications of Biosynthesized Gold Nanoparticles from Cyanobacteria: an Overview.

Recently there had been a great interest in biologically synthesized nanoparticles (NPs) as potential therapeutic agents. The shortcomings of conventional non-biological synthesis methods such as generation of toxic byproducts, energy consumptions, and involved cost have shifted the attention towards green syntheses of NPs. Among noble metal NPs, gold nanoparticles (AuNPs) are the most extensively used ones, owing to the unique physicochemical properties. AuNPs have potential therapeutic applications, as those are synthesized with biomolecules as reducing and stabilizing agent(s). The green method of AuNP synthesis is simple, eco-friendly, non-toxic, and cost-effective with the use of renewable energy sources. Among all taxa, cyanobacteria have attracted considerable attention as nano-biofactories, due to cellular uptake of heavy metals from the environment. The cellular bioactive pigments, enzymes, and polysaccharides acted as reducing and coating agents during the process of biosynthesis. However, cyanobacteria-mediated AuNPs have potential biomedical applications, namely, targeted drug delivery, cancer treatment, gene therapy, antimicrobial agent, biosensors, and imaging.

Lycopene: A Natural Arsenal in the War against Oxidative Stress and Cardiovascular Diseases.

Lycopene is a bioactive red pigment found in plants, especially in red fruits and vegetables, including tomato, pink guava, papaya, pink grapefruit, and watermelon. Several research reports have advocated its positive impact on human health and physiology. For humans, lycopene is an essential substance obtained from dietary sources to fulfil the body requirements. The production of reactive oxygen species (ROS) causing oxidative stress and downstream complications include one of the major health concerns worldwide. In recent years, oxidative stress and its counter strategies have attracted biomedical research in order to manage the emerging health issues. Lycopene has been reported to directly interact with ROS, which can help to prevent chronic diseases, including diabetes and neurodegenerative and cardiovascular diseases. In this context, the present review article was written to provide an accumulative account of protective and ameliorative effects of lycopene on coronary artery disease (CAD) and hypertension, which are the leading causes of death worldwide. Lycopene is a potent antioxidant that fights ROS and, subsequently, complications. It reduces blood pressure via inhibiting the angiotensin-converting enzyme and regulating nitrous oxide bioavailability. It plays an important role in lowering of LDL (low-density lipoproteins) and improving HDL (high-density lipoproteins) levels to minimize atherosclerosis, which protects the onset of coronary artery disease and hypertension. Various studies have advocated that lycopene exhibited a combating competence in the treatment of these diseases. Owing to all the antioxidant, anti-diabetic, and anti-hypertensive properties, lycopene provides a potential nutraceutical with a protective and curing ability against coronary artery disease and hypertension.

Glycerol-based NaDES as green solvents for ultrasound-assisted extraction of phycocyanin from Arthrospira platensis—RSM optimization and ANN modelling

Phycocyanin, a natural blue phycobiliprotein extracted from cyanobacteria, recently focused the interest of food industry as promising pigment and antioxidant. Eco-extraction of phycocyanin via ultrasound using five glycerol-based NaDES was described here. Response surface methodology (RSM) and artificial neural network (ANN) were adapted for the generation of predictive models to simulate and optimize phycocyanin extraction. Input parameters were extraction time (15–60 min), temperature (25–50 °C) and water addition (20–60%). Two original glycerol-based NaDES, glycerol:glucose:betaine (4:1:1) and glycerol:glucose:proline (4:1:1), displayed the highest phycocyanin recovery, combined with high free radical scavenging activity. Additionally, a stabilizing effect of NaDES on phycocyanin was highlighted after accelerated ageing. Optimized extraction protocol developed here allowed then the extraction, pre-formulation and stabilization of phycocyanin in one step, opening the way to a sustainable formulation of this bioactive pigment for food industry.

Exploring bioactive pigments from marine bacterial isolate from the Indonesian seas

Marine microbes offer a significant source for biodiscovery due to their rich biodiversity and genetic capacity. Particularly, microbial pigments of marine origin are getting more attention in current research due to their widely perceived application as natural food colorants, antioxidant, antimicrobial, and many more. In the past five years, our research group has successfully characterised various bioactive pigments isolated from marine bacteria, including Erythrobacter flavus strain KJ5 that produces unique sulphur-containing carotenoids, Pseudoalteromonas rubra strain PS1 and SB14 that contain antimicrobial prodiginine, and Seonamhaeicola algicola strain CCI for high content of zeaxanthin. This paper describes the challenges we encountered in conducting research in exploring bioactive pigments especially with focus on carotenoid research, reviewed critically on strategy we developed for isolation of isolate as well as identification and elucidation of the pigments, and consideration for future research.

Valorization of seabuckthorn pomace to obtain bioactive carotenoids: An innovative approach of using green extraction techniques (ultrasonic and microwave-assisted extractions) synergized with green solvents (edible oils)

Seabuckthorn pomace (SBT-P) valorization is gaining importance due to the presence of health promoting bioactive compounds. Designed with a green refinery approach, carotenoids (a bioactive pigment) was extracted from SBT-P using green extraction technologies (Ultrasound- and Microwave-assisted extraction, UAE & MAE), synergized with corn and olive oils used as green solvents. Total carotenoid content (TCC) in SBT-P extracts ranged between 26.91 ± 1.96 and 34.35 ± 0.94 mg/100 g in UAE & MAE. Among the different extraction techniques, HPLC data revealed UAE and olive oil extracts to exhibit highest yield of major carotenoids ( lycopene 87.0; β--carotene 18.7 mg/L). The percent inhibition of DPPH free radical ranged between 92.49 ± 1.84 and 94.16 ± 1.06%, whereas total phenolic content ranged from 341.02 ± 10.97 to 405.58 ± 9.58 mg gallic acid equivalent (GAE)/g of extract in UAE and MAE, respectively. Results encourages the use of green technologies for extraction of bioactive carotenoids from SBT-P and are expected to find potential food industrial applications, where natural pigments are in demand.

Optimization of fermentation time and grain concentration for water kefir production from butterfly pea flower (Clitoria ternatea)

Herbal tea is known rich in bioactive compounds and pigments. Herbal tea can be derived from Butterfly pea flowers. Butterfly pea has a high level of anthocyanins and flavonoid levels. However, butterfly pea has not been used optimally in fermentation products such as water kefir. Water kefir has many benefits and potential antioxidant sources. This research aimed to optimize fermentation time (24-48 hours) and concentration grain (5-15%) for the production of butterfly pea water kefir with a good antioxidant activity using response surface methodology. Central Composite Design was used to develop models of the response. The response used in this research is antioxidant activity, flavonoid, anthocyanin and brightness. The optimum condition was obtained at 36 hours 2 minutes of fermentation time, and 15% of grain concentration using Design Expert 10.01. The optimum combination for butterfly pea water kefir resulted in 58 ppm of antioxidant activity, 39 of brightness, 81 mg/g QE of flavonoid and 4.2 ppm of anthocyanin. A validation experiment showed that no significant difference between actual and predicted values based on paired t-test (p>0.05) using Minitab 17.0.

Screening of Bio-active Pigments, Antioxidant Activity, Total Phenolic and Flavonoid Content of Some Economically Important Medicinal Plants for Ethno-botanical Uses

An experiment was conducted to determinate the phytochemical properties (chlorophyll-a, chlorophyll-b, carotenoids, antioxidant activity, total phenolic and flavonoid content) in ten medicinal plants namely Syal Kata (Argemone mexicana L.), Akanda (Calotropis gigantea (L.) Dryand.), Dumur (Ficus carica L.), Chalmugra (Gynocardia odorata R.Br.), Kata kachu (Lasia spinosa (L.) Thwaites), Shetodrone (Leucas aspera (Wild.)), Khona (Oroxylum indicum (L.) Kurz), Reri (Ricinus communis L.), Ghat kachu (Typhonium trilobatum (L.) Schott) and Bazna (Zanthoxylum rhetsa DC.) for ethno-botanical uses. The young fresh leaves were harvested and were subjected to methanolic (95%) extract. Total phenolic contents were analysed by using Folin - Ciocalteau method where gallic acid was used as standard. Total phenolic content varied from 94.53 mg GAE/100 g FW (C. gigantea) to 484.88 mg GAE/100 g FW (L. aspera). Total flavonoid contents were performed by using Quercetin as standard. Total flavonoid content varied from 137.25 mg QUE /100 g FW (C. gigantea) to 334.27 mg QUE/100 g FW (G. odorata). Antioxidant activity of these extracts was performed by using DPPH free radical scavenging assay. Total anti-oxidant capacity varied from IC50 value 35.37 mg/mL (A. mexicana) to 90.47 mg/mL (F. carica) where ascorbic acid is used as standard. Results indicated that among the ten medicinal plants phenolics rich in L. aspera, flavonoids rich in G. odorata, antioxidant activity rich in A. mexicana. Z. rhetsa leaf rich in chlorophyll-a content (311.67 mg/100 g FW), L. aspera leaf rich in chlorophyll-b content (157 mg/100 g FW), Z. rhetsa rich in chlorophyll- (a+b) content as 439 mg/100 g FW, A. mexicana leaf rich in carotenoids content as 96 mg/100 g FW.

Cytotoxic Properties, Anthocyanin and Furanocoumarin Content of Red-Pigments Obtained from Callistemon citrinus (Curtis) Skeels Flowers

Background and objectives: There is growing interest in introducing safe and bioactive natural red pigments to the pharmaceutical and cosmetic industries. This study was designed to determine the phytochemical content and potential cytotoxicity of red pigment from Callistemon citrinus (Curtis) Skeels (syn. Melaleuca citrina (Curtis) Dum.Cours.) flowers. Methods: The flowers’ anthocyanin rich pigment was extracted with ethanol (70%, v/v) containing 0.5% formic acid. This extract was fractionated by a three-step process through a Sep-Pak C18 cartridge with water, ethyl acetate, and methanol, respectively. The anthocyanin and coumarin content of the red pigment was identified based on a reverse phase high performance liquid chromatography-electrospray ionization mass spectrometry (HPLC/ESI-MS/MS). The MTT assay was used to assess the cytotoxicity of red pigment evaluated on normal human foreskin fibroblasts as well as two malignant cell lines: human breast cancer (MCF-7) and human fibrosarcoma cells (HT1080) at 24, 48 and 72 hours. Results: The anthocyanin compounds in the red pigment fraction were cyanidin (1), cyanidin 3,5-O-diglucoside (2), cyanidin 3-O-glucoside (3), and cyanidin 3-O-glucoside-8-ethyl-catechin (4). Flowers’ pigment also contained two furanocoumarins, including 8-(but-2-en-2-yl)-8,9-dihydro-2H-furo[2,3-h] chromen-2-one (or 15-methyl angenomallin, (5)) and 9-methyl-7H-furo[3,2-g] chromen-7-one (or 8-methylpsoralen, (6)). According to the MTT assays, the highest cytotoxic effect was observed on human foreskin fibroblasts with an IC50 values of <12.5 (µg/ mL, 24 h) and 85.2 (µg/ mL, 48 h). Conclusion: It might be assumed that application of the red pigment of C. citrinus in topical formulations and cosmetics should be done with caution due to the observed cytotoxicity on dermal fibroblasts.

Identification and Verification of the Prodigiosin Biosynthetic Gene Cluster (BGC) in Pseudoalteromonas rubra S4059.

ABSTRACTPseudoalteromonas rubra S4059 produces the red pigment prodigiosin, which has pharmaceutical and industrial potential. Here, we targeted a putative prodigiosin-synthesizing transferase PigC, and a pigC in-frame deletion mutant did not produce prodigiosin. However, extractions of the pigC mutant cultures retained antibacterial activity, and bioassay-guided fractionation found antibacterial activity in two fractions of blue color. A precursor of prodigiosin, 4-methoxy-2,2′-bipyrrole-5-carbaldehyde (MBC), was the dominant compound in both the fractions and likely caused the antibacterial activity. Also, a stable blue pigment, di-pyrrolyl-dipyrromethene prodigiosin, was identified from the two fractions. We also discovered antibacterial activity in the sterile filtered (nonextracted) culture supernatant of both wild type and mutant, and both contained a heat-sensitive compound between 30 and 100 kDa. Deletion of prodigiosin production did not affect growth rate or biofilm formation of P. rubra and did not change its fitness, as the mutant and wild type coexisted in equal levels in mixed cultures. In conclusion, a prodigiosin biosynthetic gene cluster (BGC) was identified and verified genetically and chemically in P. rubra S4059 and a stable blue pigment was isolated from the pigC mutant of S4059, suggesting that this strain may produce several prodigiosin-derived compounds of pharmaceutical and/or industrial potential.IMPORTANCE Pigmented Pseudoalteromonas strains are renowned for their production of secondary metabolites, and genome mining has revealed a high number of biosynthetic gene clusters (BGCs) for which the chemistry is unknown. Identification of those BGCs is a prerequisite for linking products to gene clusters and for further exploitation through heterologous expression. In this study, we identified the BGCs for the red, bioactive pigment prodigiosin using genomic, genetic, and metabolomic approaches. We also report here for the first time the production of a stable blue pigment, di-pyrrolyl-dipyrromethene prodigiosin (Dip-PDG), being produced by the pigC mutant of Pseudoalteromonas rubra S4059.

Phycocyanobilin-modified β-lactoglobulin exhibits increased antioxidant properties and stability to digestion and heating

β-lactoglobulin (BLG) is a major whey protein with numerous techno-functional properties desirable for the food industry. Phycocyanobilin (PCB), a bioactive pigment of Arthrospira platensis with health-promoting effects, covalently binds to BLG at physiological pH. This study investigated the effects of this covalent modification on BLG functional properties. The BLG–PCB adduct possesses enhanced antioxidant properties, and bound PCB protects BLG against free radical-induced oxidation. Despite the similar thermal stabilities of BLG and BLG–PCB, BLG–PCB is less susceptible to covalent and noncovalent aggregation under moderate heat treatment (63 °C, 30 min). Blocked thiol group and reduced hydrophobicity due to hindering of hydrophobic residues by bound PCB, as well as the heat-induced transition of β-sheet to α-helix, contributed to the low susceptibility of BLG–PCB to aggregation. BLG–PCB has a higher resistance to pepsin and pancreatin digestion than BLG and unaltered IgE-binding properties. The improved functional properties of BLG–PCB make it a useful ingredient in the food industry.

Integrative analysis of metabolome and transcriptome reveals the mechanism of color formation in white root (Salvia miltiorrhiza)

Tanshinones are a class of bioactive components and pigments found in the traditional Chinese herb Salvia miltiorrhiza (S. miltiorrhiza). To clarify the biosynthesis of tanshinones in S. miltiorrhiza roots, the metabolome and transcriptome were compared among three S. miltiorrhiza lines (two red root lines: HG and CK, one white root line: BG). In this study, 18 tanshinones showed markedly lower abundances in BG lines than in other lines and 15 significant differentially accumulated metabolites (DAMs) were chosen as potential marker compounds. Meanwhile, the expression of 5 tanshinone biosynthesis genes in BG was significantly downregulated, which affected the metabolic flow of tanshinones. We speculated that this was the reason BG exhibited white roots. Moreover, 49 cytochrome P450 genes and 22 transcription factors were identified as candidate genes for tanshinone synthesis and regulation. Our results not only explain the formation of root color in S. miltiorrhiza, but also provide an effective solution to explore the biosynthetic pathway of tanshinones.

Development of hull-less barley with ultra-low gluten content via target genes combination. I. Isolation of triple mutants and black grained genotypes

Aim. The purpose of the research, presented in this paper, is to develop (as the first one in Ukraine) hull-less barley breeding material with ultra-low gluten, combining three hordein-deficient mutations, derived from Risø56, Risø1508, and R118, with black grain color as an indication of bioactive pigments content, enhancing the functional status of grain as a food product. Methods. We used electrophoretic analysis of proteins in polyacrylamide gel, DNA isolation by the CTAB method, polymerase chain reaction with DNA-markers, restriction analysis and electrophoretic separa tion of DNA in agarose gel. Results. The article presents the results of the first stage of combining target hordeindefi cient mutations by binary crosses. The F2/3 and F3/4 populations were derived from binary crosses of Risø56 × Risø1508 (reciprocal Risø1508 × Risø56) and R118 × hull-less barley Achilles. The new PCR protocol was optimized. Markers B1hor, 3a5F and Dhor-m were selected to identify mutations in B, C and D hordeins. Conclusions. Several barley genotypes with one or two target hordein-deficient mutations and black grain color were isolated. The genotypes obtained will be used in a further backcrosses program, aimed at developing of a black hull-less barley variety with ultra-low gluten content.

Biologically Plant-Based Pigments in Sustainable Innovations for Functional Textiles – The Role of Bioactive Plant Phytochemicals

The fossil-based emissions and effluents from the textile wet processing units have acute implications on the environment and human wellbeing. Therefore, a paradigm shift towards sustainable alternative materials and technology has become imperative. Thus, the overview herein emphasizes the sustainable chemistry that ubiquitously exists to propel ecological textile processing with herbal biomaterials. The review paper succinctly accentuates the efficient qualities of bioactive plant pigments for functional textiles. The paper also outlines the sustainable method of processing with plant phytochemicals. The eco-friendly herbal materials as an excellent example of Biomimicry is presented. Likewise, the multi-disciplinary research approach is discussed herein to refute global dilemmas and impel holistic ecological outcomes. The circular materials for the circular economy are detailed.

Electrochromic Properties and Electrochemical Behavior of Marennine, a Bioactive Blue-Green Pigment Produced by the Marine Diatom Haslea ostrearia.

Marennine has long been known as the unique peculiar pigment responsible for the natural greening of oysters. It is specifically produced by the marine diatom Haslea ostrearia and it is a natural blue molecule indeed promising for food industry because of the rarity of such non-toxic, blue-colored pigments. In the search for its still not defined molecular structure, investigation of the color changes with the redox state has been carried out combining different approaches. Reducing and oxidizing chemicals have been added to purified marennine solutions and a stable blue-green color has been confirmed for the oxidized state, while a yellow color corresponded to the reduced unstable state. Raman spectroscopy has been used to monitor changes in the Raman spectra corresponding to the different colored states, and cyclic voltammetry has allowed the detection of a redox system in which protons and electrons are exchanged. These findings show that marennine is a suitable stable blue pigment for use in food applications and help in the elucidation of the chromophore structure.

Extraction of Carotenoids from Pumpkin Peel and Pulp: Comparison between Innovative Green Extraction Technologies (Ultrasonic and Microwave-Assisted Extractions Using Corn Oil).

Natural pigments improve aesthetic value as well as antioxidant potential of a food product. This study was designed to determine the effects of green extraction techniques on carotenoids, polyphenols and antioxidant activities of pulp and peel of two varieties of pumpkin (Cucurbita maxima). Innovative green extractions (IGE; Ultrasound and Microwave-Assisted Extractions) synergised with corn oil (used as green solvent) were compared with conventional extraction (CE; hexane/isopropyl alcohol; 60:40, v/v). Results showed total carotenoids to be almost double on employing IGE (PM2-UAE-peel = 38.03 ± 4.21; PM4-UAE-peel = 33.78 ± 1.76 µg/g) when compared to conventional extraction (PM2-CE-peel = 19.21 ± 4.39; PM4-CE-peel = 16.21 ± 2.52 µg/g). Polyphenolic contents ranged between 510.69 ± 5.50 and 588.68 ± 7.26 mg GAE/100 g of extract in IGE, compared with conventional extracts (269.50 ± 2.17 to 318.46 ± 6.60 mg GAE/100 g) and percent inhibition of 2,2-Diphenyl-1-picrylhydrazyl (DPPH) ranging between 88.32 ± 1.51 and 93.53 ± 0.30% in IGE when compared with conventional extraction (50.61 ± 1.44 to 57.79 ± 2.09%). Further, oxidative stability of carotenoids extracts from IGE (protection factor = 1.59 ± 0.01 to 1.81 ± 0.05) were found to be significantly higher (p < 0.05) than conventional extracts. Based on results, this study supports the use of innovative green extraction techniques to obtain bioactive pigments like carotenoids. It is anticipated that results generated will find potential applications in food, pharmaceutical and cosmetic industries.

A facile macroporous resin-based method for separation of yellow and orange Monascus pigments

The yellow Monascus pigments (YMPs) named monascin and ankaflavin and the orange Monascus pigments (OMPs) named rubropunctatin and monascorubrin are two groups of bioactive components in a mixture state in the Monascus fermented products. In order to separate these two groups of bioactive pigments, a facile macroporous resin-based method was developed. The weak-polar resin CAD-40 was selected from the seven tested macroporous resins as it revealed better properties for the adsorption and desorption of the YMPs and OMPs. Then, CAD-40 resin was used for column-chromatographic separation. After eluted by 4 bedvolumes of ethanol, the yellow group (monascin and ankaflavin) and the orange group (rubropunctatin and monascorubrin) were successfully separated and purified, with an increased content from 49.3% and 44.2% in the crude pigment extract to 85.2% and 83.0% in the final products, respectively. This method would be helpful for the large-scale separation and purification of Monascus pigment products with specific bioactivity.

Bioactive Pigments of Monascus purpureus Attributed to Antioxidant, HMG-CoA Reductase Inhibition and Anti-atherogenic Functions

Monascus purpureus is known to produce pigment molecules. The pigments were extracted from M. purpureus fermented rice. In-vitro antioxidant effects of pigments were observed and presumed to alleviate oxidative stress related atherosclerosis effect in rats fed with high fat diet (HFD) for 14 weeks. The formation of lipid peroxide due to the oxidation of serum lipid was higher in rats fed with HFD. While, the feeding of fermented rice (groups III-V) significantly lowered the formation of lipid peroxide (27.1–51.7%) in serum of rats, indicated antioxidative effect of pigments. In addition, feeding of fermented rice lowered serum cholesterol and triacylglycerol by 44.82 and 45.30%, respectively. Whereas, LDL-cholesterol levels were decreased by 70.12% and HDL-cholesterol increased by 34.58%. The atherogenic indices (LDL/HDL and TC/HDL) were reduced by 77.80 and 61.05%, respectively, in rats fed with fermented rice. These data confirmed the anti-atherosclerotic effect of pigments. Further liver enzyme, 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase activity was significantly inhibited up to 54%. The identification of statins, sterols and fatty acids in fermented rice revealed the HMG-CoA reductase inhibitory activity. This was confirmed by synthesis of lower levels of cholesterol and triacylglycerol in liver of rats fed with fermented rice. Accordingly antioxidant, inhibition of HMG-CoA reductase, anti-atherogenic functions of M. purpureus fermented rice is attributed to the collective effect of bioactive metabolites.