Carotenoid Yield Research Articles

Effects of pre-treatments on the yield and carotenoid content of Gac oil using supercritical carbon dioxide extraction

The effects of air-drying temperature, particle size and enzymatic pre-treatment on the oil yield and content of carotenoids in the resultant Gac oil using supercritical CO2 extraction method were investigated. It was found that the highest oil yield was obtained when using the enzyme concentration at 0.1% (w/w) for pretreatment before air-drying. Furthermore, the content of carotenoids was also enhanced by suitable drying temperature and the particle size. High oil yield (34% g/g) and high content of β-carotene (83mg/100mL oil) and lycopene (508mg/100mL oil) were obtained using the drying temperature of 50°C and particle size of 0.45mm as pre-treatments prior to SC-CO2 extraction at pressure of 200bar and extraction temperature of 50°C. Results also indicated that the most suitable extraction time was 120min. It was concluded that Gac oil containing high amount of carotenoids could be extracted using the chosen air-drying temperature, particle size, enzymatic pre-treatment and extraction time.

Characterization of a new zeaxanthin producing strain of Chlorella saccharophila isolated from New Zealand marine waters

A fast growing strain of Chlorella saccharophila was isolated from the marine water of New Zealand and grown in heterotrophic conditions using glucose or glycerol as a carbon source. Biomass production was found to be higher in culture fed with glucose (2.14±0.08 g L(-1)) as compared to glycerol (0.378±0.04 g L(-1)). Lipid accumulation was similar for both carbon sources, at approximately 22% of dry cell weight. However, carotenoid yield was higher with glycerol (0.406±0.0125 mg g(-1)) than with glucose (0.21±0.034 mg g(-1)). Further optimization of the growth of the isolate gave maximal carotenoid production of 16.39±1.19 mg g(-1) total carotenoid, containing 11.32±0.64 mg g(-1) zeaxanthin and 5.07±0.55 mg g(-1) β-carotene. Comparison of various chemical and physical carotenoid extraction methods showed that ultrasonication was required for maximum extraction yields. The new strain has potential for biofuel, with carotenoid co-production.

Wet‐plate culture studies of Penicillium sp. PT95 and Q1 for mass production of sclerotia

Penicillium sp. PT95 and Q1 strains were able to form abundant orange, sand-shaped sclerotia in which carotenoids were accumulated. To determine the potential availability of the wet-plate method for mass production of sclerotia, nine kinds of liquid media were used culture the PT95 and Q1 strains. The results of the wet-plate culture showed that on 25% glycerol nitrate broth medium, the growth of both strains was relatively slow, and no sclerotia were found. Q1 strain cultured on Czapek's yeast extract broth medium could not form sclerotia. On other media, both strains could form sclerotia. For PT95 strain, the highest sclerotial biomass (380 mg plate(-1) ) and carotenoids yield (20.88 µg plate(-1) ) could be obtained on Czapek's yeast extract broth and Georgiou's liquid medium, respectively. For Q1 strain, malt extract broth medium gave the highest sclerotial biomass (340 mg plate(-1) ) and omitting iron Joham's liquid medium gave the highest carotenoids yield (18.29 µg plate(-1) ). The results from this study suggest the potential usage of wet-plate method in the mass production of sclerotia of the PT95 and Q1 strains.

Production of biomass, carotenoid and other lipid metabolites by several red yeast strains cultivated on waste glycerol from biofuel production – a comparative screening study

This study was focused on a comparison of growth and production properties of seven red yeast strains of the genus Rhodotorula, Sporobolomyces and Cystofilobasidium cultivated on glycerol substrate. Production of enriched yeast biomas and specific yeast metabolites (carotenoids, ergosterol, lipids) was evaluated on medium with glucose, pure technical glycerol and/or waste glycerol from biofuel production (40 g/L) and mixture of glycerol and glucose (1:3, 1:1, 3:1; C/N ratio 57 in all cultivations). All tested strains were able to utilize glycerol as the only carbon source. Production of biomass on waste glycerol was in most strains higher than in control as well as in medium with pure technical glycerol and reached 15.97–21.76 g/L. Production of carotenoids and ergosterol was better in glucose medium than in medium with glycerol only. Nevertheless, using glycerol medium with addition of glucose, higher yields of total carotenoids, beta-carotene and ergosterol were obtained than in control. The highest yields of total pigments were reached by Sporobolomyces roseus (3.60 mg/g cell dry weight (CDW); glycerol:glucose 1:3), Sporobolomyces salmonicolor (2.85 mg/g CDW; glycerol:glucose 1:3) and Rhodotorula glutinis (2.80 mg/g CDW; glycerol:glucose 3:1) In glucose medium, most tested strains except Cystofilobasidium capitatum (22.6 %) produced neutral lipids in the range of 11–15 %. Production of triacylglycerols in all strains was in 10–30 % better in glycerol medium, in which Rhodotorula aurantiaca and Sporobolomyces shibatanus also reached intracellular triacylglycerol concentrations up to 20 % of biomass. This study has shown that oleaginous red yeasts could have great potential for converting crude glycerol to valuable lipids and carotenoids in respect of efficient bioresources utilization.

Lipid and carotenoid production by oleaginous red yeast Rhodotorula glutinis cultivated on brewery effluents

This study investigated the production of microbial lipids for biodiesel production and high-value carotenoids by Rhodotorula glutinis combined with the use of brewery wastewater as carbon source for three treatments: (raw wastewater) WWraw, (glucose supplemented raw wastewater) WWglu and a (synthetic sugar medium) WWsynth. The collected brewery effluents showed high contents of sugars (maltose 24.34 g L−1; glucose 5.77 g L−1), but the low utilization of maltose led to a limitation of carbon in WWraw and WWglu. Since nitrogen was still available, carbon was channeled into cell growth instead of lipid formation, reaching an overall biomass production of 5.22 g L−1, 7.38 g L−1, and 9.55 g L−1, respectively. Carotenoids were synthesized in all treatments with total average carotenoid contents between 0.6 and 1.2 mg L−1 and with high proportions of β-carotene (∼50%) in the wastewater treatments. Suboptimal culture conditions (pH; aeration) have been identified as obstacles for higher lipid and carotenoid yields. Nevertheless, brewery wastewaters can be considered as carbon source for microbial fermentation, since they can be assumed to be an adequate source of nitrogen and other nutrients, whereas the utilization of maltose needs to be increased to achieve considerable amounts of lipid and carotenoid production.

Optimization of clean extraction methods to isolate carotenoids from the microalga Neochloris oleoabundans and subsequent chemical characterization using liquid chromatography tandem mass spectrometry

A novel experimental design was used to optimize the extraction of carotenoids from Neochloris oleoabundans using pressurized liquid extraction with food-grade solvents such as ethanol and limonene. Experimental factors, including the extraction temperature and the solvent composition, were optimized using a three-level factorial design. The response variables extraction yield and total amount of carotenoids were assessed. The statistical analysis of the results provided mathematical models to predict the behavior of the responses as a function of the factors involved in the process. The optimum conditions predicted by the model developed in this study were 112 °C as the extraction temperature and 100% ethanol as the extraction solvent. Chemical characterization of the extracts obtained was performed by means of high-performance liquid chromatography-tandem mass spectrometry. The results obtained demonstrated that, under certain growth conditions (photoautotrophically cultured in a medium supplemented with 0.3 g L(-1) KNO3), N. oleoabundans accumulated significant total amounts of the carotenoids (from 57.4 to 120.2 mg carotenoids per gram of extract depending on the extraction conditions), mainly lutein, cantaxanthin, zeaxanthin, and astaxanthin monoesters and diesters.

Photosynthesis, energy partitioning, and metabolic adjustments of the endangered Cistaceae species Tuberaria major under high temperature and drought

In view of predicted climatic changes for the Mediterranean region, study of high temperature and drought impacts on physiological responses of endangered species regains relevance. In this context, micropropagated plants of Tuberaria major, a critically endangered species, endemic of Algarve, were transferred to a controlled-environment cabinet with day/night temperatures set at 25/18°C (Reference) or 32/21°C (HT). After 15 days of HT acclimation, some plants were subjected to progressive drought followed by rewatering. The enhancement of temperature alone did not affect water relations and photosynthetic rates (PN) but the stomatal conductance (gs) exhibited a 3-fold increase in comparison with reference plants. The maximum quantum yield of photosystem (PS) II (Fv/Fm), the effective quantum yield of PSII photochemistry (ΦPSII), carotenoid (Car) and anthocyanin content enhanced, whereas the quantum yields of regulated (ΦNPQ) and nonregulated (ΦNO) energy dissipation decreased. Drought combined with HT reduced predawn leaf water potential to values of about −1.3 MPa, which had adverse effects on gas exchange and PSII activity. Values of PN and gs were 71 and 79% lower than those of HT plants. An impairment of photochemical activity was also observed: the decrease in ΦPSII and the increase of ΦNPQ. However, an irreversible photoinhibitory damage had not occurred. Carotenoid and anthocyanin content remained elevated and soluble sugars (SS) increased twice, whereas proline and MDA accumulation was not detected. On the first 24 h after water-stress relief, gs, PN, ΦPSII, and ΦNPQ did not recover, but SS returned to the reference level. Overall, T. major acquired an adequate capacity for a protection against the development of oxidative stress during drought and water recovery under HT. These findings suggest that T. major is prepared to deal with predicted climate changes.

Supercritical fluid extraction of carotenoids and chlorophylls a, b and c, from a wild strain of Scenedesmus obliquus for use in food processing

Faster and more selective extraction methods are urged to recover food-grade pigments and antioxidants from microalgae – which do not resort to food-incompatible and environment-unfriendly solvents. Hence, this study ascertained the influence of pressure, temperature, CO2 flow rate and a polar co-solvent upon the yields of carotenoids and chlorophylls in supercritical fluid extraction of Scenedesmus obliquus biomass. The highest carotenoid yield was attained at 250bar and 60°C. The yields of chlorophylls, when using plain CO2, increased slightly with pressure, but decreased with temperature and CO2 flow rate; the highest yield of chlorophyll a was at 4.3gCO2min-1, whereas ethanol as co-solvent increased all yields except that of chlorophyll c. The highest ratio of total carotenoids to chlorophyll a was reached at 250bar and 60°C. A remarkable selectivity was observed under these operating conditions, which may enable easy separation and purification of the aforementioned pigments.

Effect of modifier on the composition and antioxidant activity of carotenoid extracts from pumpkin (Cucurbita maxima) by supercritical CO2

The effects of different modifiers and processing conditions on the compositions, yield, and antioxidant activity of carotenoid by supercritical CO2 (SC-CO2) extraction were studied. Increasing the percentage of modifier would increase the yield, but no significant changes were observed on the antioxidant properties of carotenoids extracted at 50 °C with low concentration (5%) of single modifier. As single modifier, the oil showed the pronounced effects on the yield and antioxidant activity of extracts compared to the effects of water and ethanol. The combination of ethanol and water, or ethanol and oil, or water and oil in SC-CO2 extraction showed that the yields of carotenoids were higher than for the single modifier with equal amount. The modifier and operating temperature of SC-CO2 notably influenced the selectivity of carotenoids, especially the ratio of cis-β-carotene/total carotenoids. The antioxidant activity didn’t show a liner correlation with the total yield of carotenoids. The antioxidant activities of unit mass carotenoid extract were decreased after adding modifiers at both 50 °C and 80 °C. At lower concentrations of water (5% and 10%), the antioxidant activity of unit mass carotenoid extracts was higher at 50 °C. The highest antioxidant activity of unit mass carotenoid extract was found at 80 °C with both single and combined modifiers.

Progress toward an Escherichia coli canthaxanthin bioprocess

Canthaxanthin has significant commercial value due to its use as an antioxidant and feed additive in aquaculture and poultry farms. The biological production of high-value carotenoids, such as canthaxanthin, represents a significant challenge. Presented here is an Escherichia coli bioprocess, which includes the recombinant production of a strain, a novel medium and a fermentation process, for the efficient biosynthesis of canthaxanthin. The chromosomal overexpression of the E. coli genes Idi, Dxs and Dxr increased the carbon flux into the plasmid-based canthaxanthin biosynthetic pathway, achieving a total carotenoid productivity of 11.7mg/g (91% canthaxanthin). The development of a novel culture medium increased the productivity 2.9-fold to 78.2mg/L total carotenoids. This high-titer platform was subsequently employed for the analysis of mutations known to be advantageous to carotenoid biosynthesis, providing the first such evaluation in a high titer background. This demonstrated that thee increased expression of IspB or the deletion of PykF, PykA or YtjC improved the total carotenoid yield, whereas the increased expression of Pps and Ppc, was not improve productivity. This analysis resulted in an increase in the carotenoid yield to 16.1mg/g. The use of 1L lab-scale fermentations allowed for further improvements in productivity, and 170mg/L total carotenoids was obtained.

Optimization of the Conditions of Supercritical CO2 Extraction for Carotenoid from Blue Algae Using Response Surface Methodology

To optimize the conditions of supercritical CO2 extraction of carotenoid, a single factorial experiment was used to study the extracting temperature, time and pressure. A three-level Box–Behnken factorial design was used with response surface methodology (RSM) to optimize the three conditions. A mathematical model was then developed to show the effect of each factor and their interactions on the carotenoid yield. The single factorial experiment shows that the best extracting temperature, time and pressure are 50 0C,50 min and 25 MPa, respectively. The model estimates that a maximal caroteniod yield (0.624 mg/g) can be obtained when the conditions of temperature, time and pressure are set at 51.9 0C,61 min and 25.6MPa,repectively. The yield of carotenoid (0.621± 0.009 mg/g) from validation experiments agree with the predicted one.

Pressurized Hot Ethanol Extraction of Carotenoids from Carrot By-Products

Carotenoids are known for their antioxidant activity and health promoting effects. One of the richest sources of carotenoids are carrots. However, about 25% of the annual production is regarded as by-products due to strict market policies. The aim of this study was to extract carotenoids from those by-products. Conventional carotenoid extraction methods require the use of organic solvents, which are costly, environmentally hazardous, and require expensive disposal procedures. Pressurized liquid extraction (PLE) utilizes conventional solvents at elevated temperatures and pressure, and it requires less solvent and shorter extraction times. The extraction solvent of choice in this study was ethanol, which is a solvent generally recognized as safe (GRAS). The extraction procedure was optimized by varying the extraction time (2–10 min) and the temperature (60–180 °C). β-Carotene was used as an indicator for carotenoids content in the carrots. The results showed that time and temperatures of extraction have significant effect on the yield of carotenoids. Increasing the flush volume during extraction did not improve the extractability of carotenoids, indicating that the extraction method was mainly desorption/diffusion controlled. Use of a dispersing agent that absorbs the moisture content was important for the efficiency of extraction. Analysing the content of β-carotene at the different length of extraction cycles showed that about 80% was recovered after around 20 min of extraction.

Carotenoid synthesis and phytoene synthase activity during mating of Blakeslea trispora

Carotenoid formation was investigated in wild type and carotenogenic mutants of Blakeslea trispora after mating (−) and (+) strains. The highest yields of carotenoids, especially β-carotene was observed following mating. In vitro incorporation of geranylgeranyl pyrophosphate into phytoene and β-carotene corresponded to increased carotenogenesis in the mated strains. Immuno determination of phytoene synthase protein levels revealed that the amounts of this enzyme is concurrent with the increases in carotenoid content. In fungi, phytoene synthase together with lycopene cyclase are encoded by a fusion gene crtYB or carRA with two individual domains. These domains were both heterologously expressed in an independent manner and antisera raised against both. These antisera were used, to assess protein levels in mated and non-mated B. trispora. The phytoene synthase domain was detected as an individual soluble protein with a molecular weight of 40kDa and the lycopene cyclase an individual protein of mass about 30kDa present in the membrane fraction following sub-cellular fractionation. This result demonstrates a post-translational cleavage of the protein transcribed from a single mRNA into independent functional phytoene synthase and lycopene cyclase.

Supercritical Carbon Dioxide Extraction of Carotenoids from Marigold Flower Using Orthogonal Test Design

Supercritical carbon dioxide extraction had been used in the extraction conditions for carotenoids from the marigold flower (Tagetes erecta L.). Five significant factors (particle size, temperature, pressure, time and co-solvent addition) on total carotenoid yield were obtained using one-at-a-time design (p≤0.05). The optimal extraction conditions were 55 oC, 50 MPa and 180 min. Under these conditions, the yield of carotenoids was 6.872mg/g.

Carotenoid Value Addition of Cereal Products by Monoculture and Mixed‐Culture Fermentation of Phaffia rhodozyma and Sporobolomyces roseus

ABSTRACTCarotenoid value addition of corn whole stillage by red yeast fermentation has yielded astaxanthin‐ and β‐carotene‐enriched distillers dried grains with solubles (DDGS) for animal feed. In this study, commonly used animal feeds (rice bran, wheat bran, milo whole stillage, and soybean products) were subjected to carotenoid value addition. Phaffia rhodozyma and Sporobolomyces roseus monoculture and mixed‐culture submerged fermentation of these substrates supplemented with 5% glycerol were analyzed for astaxanthin, β‐carotene, and residual glycerol. Among all the substrates, full‐fat rice bran and full‐fat soy flour resulted in the highest astaxanthin (80 μg/g by P. rhodozyma) and β‐carotene yields (836 μg/g by S. roseus). P. rhodozyma produced the highest astaxanthin yield on each substrate, whereas depending on the substrate, either the mixed culture or S. roseus monoculture produced the highest β‐carotene yield. Soy hull was a poor substrate for carotenoid value addition. Both yeasts used glycerol as a carbon source for carotenoid production. This study shows that substrates influence the carotenoid yield. However, it is impossible to dissect the effect of specific nutrients on carotenoid production in complex biological substrates. Carotenoid value addition of these substrates provides as much as or more than the required daily dosage of carotenoids in animal feed.

Use of waste chicken feathers as peptone for production of carotenoids in submerged culture of Rhodotorula glutinis MT-5

Peptones are one of the most expensive constituents of fermentation media. The present study was performed to prepare the peptone from waste chicken feathers through acid hydrolysis and to investigate the usability of this peptone as substrate for biomass and carotenoid production by Rhodotorula glutinis MT-5. Chicken feather peptone (CFP) was found to be rich in ash (42.1 g/100 g), protein (55.8 g/100 g) and mineral contents. The ability of CFP to support biomass and carotenoid production in the yeast was comparable to those of two commercial peptones (Tryptone peptone = TP and Fish peptone = FP). The optimum concentration of CFP was found to be 8 g/L for both biomass and carotenoid production. Adding 8 g/L CFP to the medium increased carotenoid and biomass production about 53 and 36% compared to control, respectively. At the end of the fermentation, the maximum biomass (14.2 g/L) and carotenoid (92 mg/L) concentrations were achieved with CFP. Moreover, the maximum carotenoid yield (6.47 mg/g) was reached in CFP medium. This study showed for the first time that waste chicken feathers could be effectively used as a novel carotenoid production substrate for R. glutinis.

OPTIMIZATION OF SALT CONCENTRATIONS FOR A HIGHER CAROTENOID PRODUCTION IN DUNALIELLA SALINA (CHLOROPHYCEAE)(1).

Dunaliella salina (Dunal) Teodor, when treated over 25 d with a wide range of NaCl salinities (0.6-4.5 M), showed its maximal growth potentialities at 1.5-3.0 M NaCl and was able to survive even at 4.5 M NaCl. Sodium concentrations increased significantly at the supraoptimal salinities, reaching up to 5 mmol · g(-1) dry weight (dwt) at 4.5 M NaCl. Interestingly, ability of D. salina to take up essential mineral nutrients was not impaired by increased salinity. As for growth, chl concentrations were maximal in the 1.5-3.0 M NaCl range. Interestingly, carotenoid concentrations increased with the increasing salinity. The highest values of total antioxidant activity (5.2-6.9 mg gallic acid equivalents [GAE] · g(-1) dwt), antiradical activity, and reducing power were measured at 1.5-3.0 M NaCl. As a whole, these results showed that at 1.5-3.0 M NaCl, D. salina produce appreciable antioxidant level. But, once it reaches its growth maximum, a salt addition up to 4.5 M could enhance its carotenoid yield.

Extraction, Characterization, Antioxidant and Anti-Inflammatory Properties of Carotenoids from the Shell Waste of Arabian Red Shrimp Aristeus alcocki, Ramadan 1938

Shrimp processing waste is the single largest industrial waste in India causing diverse environmental problems. Extraction of carotenoids from the shell waste of the Arabian red shrimp Aristeus alcocki was investigated using different organic solvents, and vegetable oils, under wet and dry conditions, with and without deproteinization. The highest carotenoid yield was obtained with non-deproteinized wet waste extracted using acetone. The carotenoid yield was found to be double that of Pandalus borealis shell waste, which is currently used as one of the commercial sources of natural astaxanthin. Thin Layer Chromatography (TLC) analysis of the carotenoid extract showed that it contains free astaxanthin, astaxanthin monoester and astaxanthin diester in the ratio 1:1:2. gas chromatography (GC) analysis of the fatty acids esterified with astaxanthin revealed that saturated fatty acids, monounsaturated fatty acids and poly unsaturated fatty acids (PUFAs) are in the ratio 5:3:2 in monoester, whereas in diester they are in the ratio 4:3:3. The main fatty acids in monoester and diester are palmitic acid, oleic acid, stearic acid and � -3 PUFAs: docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA). The in vitro antioxidant activity of the carotenoid extract showed significant hydroxyl radical scavenging activity, superoxide anion scavenging activity and inhibition of lipid peroxidation at nanogram levels. The carotenoid extract significantly reduced carageenan induced paw edema in mice, percentage inhibition being 47.83 and 67.11 percent at astaxanthin concentrations of 0.5 mg/kg body weight and 1.0 mg/kg body weight, respectively. The inhibition of inflammation at 1.0 mg/kg body weight was greater than that produced by the standard reference drug diclofenac. The strong antioxidant and anti-inflammatory activities exhibited by the carotenoid extract of the shrimp shell might be due to the combined action of astaxanthin and � -3 PUFAs present in the astaxanthin esters.

A potent lipid producing isolate of Epicoccum purpurascens AUMC5615 and its promising use for biodiesel production

Oils of oleaginous microorganisms are a powerful alternative to vegetable oils for biodiesel production. In this study, the fungus Epicoccum purpurascens AUMC5615 isolated from Egypt showed a potent high lipid content (80% lipid) when grown on 4% sucrose in submerged culture under continuous illumination. Under dark submerged conditions the lipid content has drastically decreased to 12%. In light static conditions, the lipid content was 70%; however, the net lipid yield was significantly lower than that of light submerged cultures because of the decrease in growth under light static conditions in comparison to light submerged cultures. Under dark static conditions the lipid content of the fungus has declined to 30%. These results indicate that light plays a crucial role in the lipid accumulation whereas submersion enhances the growth of the fungus. Concomitantly, the highest yield of carotenoids was obtained under light submerged conditions followed respectively by light static, dark submerged and dark static. This synchronized increase in carotenoids content might be implicated in protecting the high lipid pools in the fungus from peroxidation. Growing the fungus on 4% of crude molasses resulted in a net lipid production of 26.8 g per liter under light submerged conditions. The determination of fatty acids by GC/MS revealed that the major constituents are four saturated fatty acids, hexadecanoic, n-decanoic, dodecanoic and octadecanoic acids. These saturated fatty acids would give valuable stability properties of such fungal biodiesel. The current investigation opens the scope for the possible use of this promising fungal isolate in biodiesel production.

Process optimisation for recovery of carotenoids from tomato waste

Carotenoids constitute an important component of waste originating from tomato processing plants. Studies were carried out to assess the extraction yield of tomato waste carotenoids in different solvents and solvent mixtures and to optimise the extraction conditions for maximum recovery. A mixture of ethyl acetate and hexane gave the highest carotenoid extraction yield among the others examined. Extraction conditions, such as percentage of hexane in the solvent mixture of ethyl acetate and hexane, ratio of solvent to waste and particle size were optimised using a statistically designed experiment. A regression equation for predicting the carotenoid yield as a function of three extraction variables was derived by statistical analysis and a model with predictive ability of 0.97 was obtained. The optimised conditions for maximum carotenoid yield (37.5mgkg−1drywaste) were 45% hexane in solvent mixture, solvent mixture to waste ratio of 9.1:1 (v/w) and particle size 0.56mm.