Extraction of chlorophylls and carotenoids from dry and wet biomass of isolated Chlorella Thermophila: Optimization of process parameters and modelling by artificial neural network

Chlorophylls and carotenoids can be extracted from microalgae using various solvents. However, there is lack of studies regarding the comparison of extraction yield of these pigments from wet and dry microalgal biomass using different combination of cell disruption methods. Therefore, in this work, we have investigated the comparison of the extraction yield of chlorophylls and carotenoids from the wet and heat-dried microalgal biomass (isolated Chlorella thermophila) using ethanol. Extraction parameters such as homogenisation time, homogenisation speed, solvent temperature, solid-solvent ratio, boiling time and microwave time have been optimised. Chlorophyll extraction yield was observed to be 2.7 fold higher from wet biomass than dry biomass while carotenoid yield was 6.7 fold higher. Highest chlorophyll yield (∼60 mg/g-dry biomass) was observed at 6 min of homogenisation time, 10,000 rpm, solid solvent ratio of 1 mg/mL and 58 °C of solvent temperature from wet biomass with extraction efficiency of ∼94 %. Highest carotenoid yield was noticed following the same conditions of chlorophyll extraction except 4 °C of solvent temperature. The modelling of the extraction process was performed using artificial neural network (ANN) which may be useful for the scale-up of the extraction process at the industrial level.     

Recovery of carotenoids from ensilaged shrimp waste

A method for fermentation of shrimp waste was standardized using a statistically designed experiment, with respect to three variables namely, levels of glucose and starter culture and time of fermentation. The optimized levels for achieving the desired pH was 20.5% glucose, 19.5x10(4)cells/g of starter culture and fermentation time of 70h. Recovery of carotenoids from fermented and acid ensiled shrimp waste was assessed during 75 days of storage. Acid ensilaging resulted in the reduction of solvent extraction yield of carotenoids from 43.09 to 26.76 microg/g by the end of 75 days of storage. The yield of oil extracted carotenoids was higher in both types of silage at the end of 75 days storage compared to the initial yield, being 31.30 microg/g in fermented silage and 26.18 microg/g in acid silage. The results indicated the usefulness of fermentation as a method for stabilization and recovery of carotenoids in the shrimp waste.     

Efficient and environmentally friendly method for carotenoid extraction from Paracoccus carotinifaciens utilizing naturally occurring Z-isomerization-accelerating catalysts

This study aimed to improve the efficiency of Paracoccus carotinifaciens-derived carotenoid (astaxanthin, adonirubin, adonixanthin) extraction using environmentally friendly Z-isomerization-accelerating catalysts. Adding naturally occurring catalysts such as isothiocyanates and polysulfides to the extraction solvent significantly improved the efficiency of carotenoid extraction, likely because of enhanced solubility of carotenoid Z-isomers compared with all-E-isomers. Indeed, addition of the catalysts markedly increased the content of carotenoid Z-isomers in the resulting extract. The use of a catalyst allyl isothiocyanate, which is abundantly included in Brassicaceae plant family, at high extraction temperature and long extraction time led to increased carotenoid recovery and Z-isomer content. These findings will enhance the efficiency of organic solvent-based extraction of carotenoids from carotenoid-rich sources. Numerous studies have reported that the Z-isomers of carotenoids exhibit greater bioavailability and antioxidant capacity than the all-E-isomers. Hence, the method proposed here utilizing Z-isomerization-accelerating catalysts could enhance both the extraction efficiency and beneficial health effects of carotenoids.     

An Autolytic Process for Recovery of Antioxidant Activity Rich Carotenoprotein from Shrimp Heads

Studies were carried out to utilize in situ proteases of shrimp heads to recover carotenoproteins possessing antioxidant activity. Highest protease activity of the buffer extract was found at pH 8.0 (9.85 ± 0.61 units). The protease activity increased with temperature up to 50°C and reduced thereafter with highest activity being 19.32 ± 2.0 units. Thus, the autolysis of shrimp heads for recovery of carotenoprotein was carried out at pH 8.0 and at 50°C. Waste to buffer ratio had a significant (p < 0.05) effect on recovery of carotenoids in carotenoprotein filtrate with a maximum of 58.5 ± 6.4% recovery with a waste to buffer ratio of 1:2.5 (w:v). The carotenoid recovery increased significantly to 63.4% ± 3.6% at the end of a 4-h autolysis. The studies on combined effect of waste to buffer ratio and autolysis time indicated increase in protein recovery with increase in waste to buffer ratio but not with autolysis time. DPPH scavenging activity of the carotenoprotein isolate increased with autolysis time up to 100 min, and thereafter, reduced above 160 min of autolysis time. With increase in waste to buffer ratio, the scavenging activity increased, reaching more than 12.5 mg TBHQ equivalent/mg protein at waste to buffer ratio of 1:5. The optimum autolysis condition for obtaining antioxidant activity rich carotenoprotein from shrimp heads was found to be waste to buffer (pH 8.0) ratio of 1:5 and an autolysis time of 2 h at 50°C. The isolated carotenoprotein was found to have antioxidant activity with respect to singlet oxygen quenching, reducing power and metal chelating activity.     

Identification and quantification of carotenoid pigments during the embryonic development of the freshwater shrimp Macrobrachium olfersii (crustacea,decapoda)

The principal carotenoids present during the 16‐day embryonic development period and in the first post‐embryonic stage (zoea I) were identified and quantified in the diadromous, neotropical, freshwater shrimp Macrobrachium olfersii, employing differential extraction, absorption spectra analysis and thin layer chromatography. The characteristic crustacean pigments α‐ and β‐carotene and astaxanthin, and a mixture of four unidentified yellow pigments were found. Total carotenoids increase significantly at the mid‐point of embryogenesis (day 8), declining to a minimum in the recently hatched first zoea. This decrease appears to reflect the establishment of carotenoid catabolism, or of carotenoid use in protein and membrane formation and stabilization towards the end of embryogenesis, as well as the depletion of yolk protein reserves by the non‐feeding first zoea.     

葵花籽油的酶辅助压榨制备工艺优化

葵花籽是一种优质的油料资源,是世界第四大油料作物。葵花籽不但含油量高,葵花籽油以其高达90%的不饱和脂肪酸和富含维生素E、胡萝卜素等营养成分的特点,被称为"保健油",有延缓衰老、调节新陈代谢和降低胆固醇等功能。采用酶辅助压榨法制备葵花籽油,在提高出油率的同时保留油脂的天然风味。通过单因素实验法研究了酶的种类、酶添加量、pH值、液固比、温度和时间对压榨葵花籽油出油率的影响,以确定最佳的工艺条件。结果表明,最佳工艺条件为：选用纤维素酶,酶添加量为0.7%,pH为4.5,液固比为25%,酶解温度为55℃,酶解时间为2.5 h。在此最佳条件下,出油率是传统冷榨法的3.48倍,油脂提取率达85%。     

Extraction of astaxanthin from giant tiger (Panaeus monodon) shrimp waste using palm oil: studies of extraction kinetics and thermodynamic

Study of extraction of astaxanthin from giant tiger (Panaeus monodon) shrimp waste using palm oil was conducted to determine the extraction kinetics and thermodynamic parameters. Two extraction models were proposed: mass transfer kinetic model and reaction kinetic model. It was found that both of mass transfer and reaction kinetic control the extraction of astaxanthin from shrimp waste using palm oil. The thermodynamic parameters of extraction were also obtained in this study.     

Applicability of Subcritical Water Treatment on Oil Seeds to Enhance Extractable Lipid

Subcritical water (SCW) has been widely studied for its unique properties both as catalyst and solvent in various chemical processes. The use of SCW to pretreat agricultural products and waste has been extensively studied for producing fermentable sugars. In this study, SCW pretreatment was carried out to increase and/or improve the extractability of oils from oil seeds like Datura stramonium, Jatropha curcas, and sunflower seeds. SCW pretreatment of D. stramonium seeds resulted in 50 % increase of oil yield (from 17.16 to 28.25 %). Although negligible increases were observed from both J. curcas and sunflower seeds, SCW pretreatment allowed full extraction of the oils without grinding and/or dehulling of the seeds. This pretreatment process caused insignificant changes in the composition and quality of the oils extracted. Efficient SCW treatment can be accomplished under mild conditions (175 °C, 3.5 MPa) in a short time (15 min).     

The carotenoids of wild and blue disease affected farmed tiger shrimp (Penaeus monodon, Fabricus)

1. The main carotenoids in wild Penaeus monodon exoskeleton were astaxanthin di- and mono-esters, astaxanthin, and beta-carotene. 2. Wild P. monodon exoskeleton contained on average 26.3 ppm total carotenoid; normally pigmented farmed shrimp had a similar concentration (25.3 ppm). 3. Exoskeletons of farmed "blue" P. monodon (i.e. blue-coloured, as opposed to the normally red-blue/black banded shrimp) contained significantly less total carotenoid (4.3-7 ppm). The only major carotenoid being astaxanthin. 4. Commercially available diets contained only trace quantities of canthaxanthin. 5. Nutritional deficiency with respect to carotenoids is suggested as the cause of blue disease in farmed P. monodon.     

Extraction of sunflower oil by twin screw extruder: screw configuration and operating condition effects

The objective of this study was to investigate the screw configuration allowing oil extraction from sunflower seeds with a twin-screw extruder. Experiments were conducted using a co-rotating twin-screw extruder. Five screw profiles were examined to define the best performance (oil extraction yield, specific mechanical energy and oil quality) by studying the influence of operating conditions, barrel temperature, screw speed and feed rate. Generally, the position and spacing between two reversed screw elements affected oil extraction yield. An increase of oil extraction yield was observed as the reversed screw elements were moved with increased spacing between two elements and with smaller pitch elements. In addition, oil extraction yield increased as barrel temperature, screw speed and feed rate were decreased. Highest oil extraction yield (85%) with best cake meal quality (residual oil content lower than 13%) was obtained under operating conditions of 120 degrees C, 75 rpm and 19 kg/h. Furthermore, the operating parameters influenced energy input. A decrease in barrel temperature and feed rate followed by an increase in screw speed increased energy input, particularly specific mechanical energy input. Effect of the operating parameters on oil quality was less important. In all experiments tested, the oil quality was very good. The acid value was below 2 mg of KOH/g of oil and total phosphorus content was low, below 100 mg/kg.     

Efficient concomitant production of lipids and carotenoids by oleaginous red yeast <Emphasis Type="Italic">Rhodotorula glutinis</Emphasis> cultured in palm oil mill effluent and application of lipids for biodiesel production

Rhodotorula glutinis TISTR 5159 is oleaginous red yeast that accumulates both lipids and carotenoids. It was cultured in palm oil mill effluent (POME) with only the addition of ammonium sulfate and Tween 20 as a suitable nitrogen source and surfactant, respectively. Response surface methodology (RSM) was applied to optimize initial chemical oxygen demand (COD) in POME, C/N ratio, and Tween 20 concentration for concomitant production of lipids and carotenoids. Among three investigated factors, C/N ratio contributed a significant effect upon lipid and carotenoids production. Analysis of response surface plots revealed that the optimum C/N ratio for the biomass was 140, while that for lipid content and carotenoids were higher at 180 and 170, respectively. The high level of the nitrogen source (with a low C/N ratio) enhanced the biomass, making the accumulation of lipids and carotenoids less preferable. Hence, the two-stage process was attempted as an optimal way for cell growth in the first stage and product accumulation in the second stage. The lipid yield and carotenoid production obtained in the two-stage process were higher than those in the one-stage process. In the semi-continuous fermentation, R. glutinis TISTR 5159 accumulated high lipid content and produced a considerably high concentration of carotenoids during long-term cultivation. Additionally, efficient COD removal by R. glutinis TISTR 5159 was observed. The biodiesel produced from yeast lipids was composed mainly of oleic and palmitic acids, similar to those from plant oil.     

Distribution of Marine Red Yeasts in Shrimps and the Environments of Shrimp Culture

Populations of marine red yeast from shrimps and the environments of shrimp culture were investigated from various areas at Zhanjiang in China. All strains were studied for the production of biomass and carotenoids. We isolated 88 marine red yeast strains and the average populations of marine red yeast in seawater and the water from shrimp culture ponds were 70.0 and 172.4 CFU per 100 ml water, respectively. For shrimp samples, average populations of marine red yeast from gills, intestines, and stomachs were 178.0, 15.0, and 8.0 CFU per shrimp, respectively. The isolates were grouped into nine species belonging to three genera as follows: Rhodosporidium, Rhodotorula, and Sporidiobolus. R. sphaerocarpum had the highest average biomass yield (10.3 ± 0.88 g/l), followed by S. ruineniae (10.1 g/l) and Rh. mucilaginosa (9.9 ± 1.75 g/l). R. paludigenum had the highest average carotenoid yield (2.83 ± 0.589 mg/l), followed by S. pararoseus (2.72 mg/l) and R. sphaerocarpum (2.59 ± 0.454 mg/l). The results showed that marine red yeasts were normal microbial components in the environments of shrimp culture and shrimps, and carotenoids are abundant in these marine red yeast.     

Efficient decomposition of shrimp shell waste using <Emphasis Type="Italic">Bacillus cereus</Emphasis> and <Emphasis Type="Italic">Exiguobacterium acetylicum</Emphasis>

Two bacterial cultures were isolated and tested for degradation of shrimp shell waste. According to morphological examination, physiological tests, and applied molecular techniques, isolates were identified as Bacillus cereus and Exiguobacterium acetylicum. Both strains were cultivated separately in flasks with 100 mL of shrimp shell waste broth (3% of washed, dried and ground shrimp shell waste in tap water, pH 7.0) at 37°C. At determined periods of time, deproteinization and demineralization of residuals were measured. Fermentation of 3% shell waste with B. cereus indicated 97.1% deproteinization and 95% demineralization. For E. acetylicum, the level of deproteinization and demineralization was 92.8 and 92%, respectively. Protein content was reduced from 18.7 to 5.3% with B. cereus and to 7.3% with E. acetylicum. No additional supplements were used during the fermentation of shell waste. B. cereus strain showed higher efficacy in decomposition of shell waste and was used for large-scale fermentation in 12 L of 10% shrimp shell waste broth. Incubation of bacteria with shell waste during 14 days at 37°C resulted in 78.6% deproteinization and 73% demineralization. High activity of isolated cultures in decomposition of shrimp shell waste suggests broad potential for application of these bacteria in environmentally friendly approaches to chitin extraction from chitin-rich wastes.     

Optimization of supercritical carbon dioxide extraction of silkworm pupal oil applying the response surface methodology

Supercritical carbon dioxide extraction (SC-CO₂) of oil from desilked silkworm pupae was performed. Response surface methodology (RSM) was applied to optimize the parameters of SC-CO₂ extraction. The effects of independent variables, including pressure, temperature, CO₂ flow rate, and extraction time, on the yield of oil were investigated. The statistical analysis showed that the pressure, extraction time, and the quadratics of pressure, extraction time, and CO₂ flow rate, as well as the interactions between pressure and temperature, and temperature and flow rate, showed significant effects on oil yield. The optimal extraction condition for oil yield within the experimental range of the variables researched was at 324.5 bar, 39.6 °C, 131.2 min, and 19.3 L/h. At this condition, the yield of oil was predicted to be 29.73%. The obtained silkworm pupal oil contained more than 68% total unsaturated fatty acids, and alpha-linolenic acid (ALA) accounted for 27.99% in the total oil.     

Carotenoid Formation by Staphylococcus aureus

The carotenoid pigments of Staphylococcus aureus U-71 were identified as phytoene; zeta-carotene; delta-carotene; phytofluenol; a phytofluenol-like carotenoid, rubixanthin; and three rubixanthin-like carotenoids after extraction, saponification, chromatographic separation, and determination of their absorption spectra. There was no evidence of carotenoid esters or glycoside ethers in the extract before saponification. During the aerobic growth cycle the total carotenoids increased from 45 to 1,000 nmoles per g (dry weight), with the greatest increases in the polar, hydroxylated carotenoids. During the anaerobic growth cycle, the total carotenoids increased from 20 nmoles per g (dry weight) to 80 nmoles per g (dry weight), and only traces of the polar carotenoids were formed. Light had no effect on carotenoid synthesis. About 0.14% of the mevalonate-2-(14)C added to the culture was incorporated into the carotenoids during each bacterial doubling. The total carotenoids did not lose radioactivity when grown in the absence of (14)C for 2.5 bacterial doublings. The total carotenoids did not lose radioactivity when grown in the absence of (14)C for 2.5 bacterial doublings. The incorporation and turnover of (14)C indicated the carotenes were sequentially desaturated and hydroxylated to form the polar carotenoids.     

Development and Quality Evaluation of Hypoallergenic Bakery Products Suitable for Patients with Wheat-associated Allergy

The influence of growth temperature (21–41°C) and light intensity on compositions of four carotenoids, and of fatty acids from carotenoid glucoside ester (carotenoid K-G-FA) and from the cellular lipids in Rhodococcus rhodochrous RNMS1 were quantitatively investigated. Lowering the temperature increased the total carotenoid content and the proportion of carotenoid K-G-FA. It increased the proportion of unsaturated fatty acids in both carotenoid K-G-FA and the cellular lipids, decreased that of saturated ones, and slightly decreased that of branched-chain ones. This bacterium adapted itself to low temperature by desaturating its fatty acids. The light intensity affected neither the content and the composition of carotenoids nor the composition of the fatty acids in carotenoid K-G-FA and the cellular lipids. This bacterium was a scotochromogenic strain.     

THE INHIBITION OF CAROTENOID ACCUMULATION IN NAVEL ORANGES BY GIBBERELLIN A3 AS MEASURED BY THIN LAYER CHROMATOGRAPHY

A study of the changes in carotenoid level in the Navel orangerind following color break supported existing evidence thatlight increases the level of carotenoids and that the periodof carotenoid accumulation ceases at a certain time, regardlessof the level of color attained. It was also shown that GA retards the accumulation of rind carotenoidsand that this effect is on general carotenoid synthesis ratherthan alteration of the ratios of individual carotenoids. A method for thin layer chromatography of carotenoids was described,using silica gel as a base. ¹ Paper No. 1562.     

Botryococcus as an alternative source of carotenoids and its possible applications – an overview

The enhanced interest in carotenoid research arises partly because of their application in the food and health industries and partly because of the necessity to find a commercially viable natural source for their mass production. The bottlenecks in finding a natural source of carotenoids which can compete with the synthetic products is the mass production of the organism that produces carotenoids, cell harvesting and extraction methods of carotenoids. The microalga Botryococcus braunii is an interesting organism for its commercial value as a rich source of carotenoids. It contains lutein as major carotenoid which is considered to be one of the beneficial carotenoids in human health applications. The current paper reviews the status of B. braunii as an alternative source of carotenoid production on the commercial scale addressing aspects like cultures of algae, factors that enhance the production and accumulation of carotenoids, cell harvesting methods, and carotenoid extraction. The paper also presents an overview of identification, characterization and structural elucidation of carotenoids from B. braunii and their bioactivity.     

Isolation of chlorophylls and carotenoids from freshwater algae using different extraction methods

Usually marine algae are an excellent source of pigments for different commercial sectors. Freshwater macroalgae can be exploited as a good source of biologically active compounds provided an appropriate extraction method is developed. The efficiency of four methods, like microwave‐assisted (MAE), ultrasound‐assisted extraction (UAE), supercritical fluid extraction (SFE) with ethanol as a co‐solvent, as well as conventional Soxhlet extraction were studied in the same conditions (time, solvent and temperature) for the recovery of chlorophylls and carotenoids from three freshwater green algae species: Cladophora glomerata, Cladophora rivularis and Ulva flexuosa. UV‐Vis spectrophotometry was used to determine chlorophyll a, chlorophyll b and total carotenoid content in obtained extracts. The results of this study showed that the advantages of novel extraction techniques (MAE and UAE) include higher yield and, in consequence, lower costs compared to traditional solvent extraction techniques. These methods were much more efficient in freshwater green algae pigment recovery than the classic Soxhlet extraction as well as SFE.     

油菜种子类胡萝卜素总量测定方法的研究

类胡萝卜素是植物中最重要的一类色素,具有抗氧化、预防心血管疾病和癌症、延缓衰老等生物学功能,与人类健康关系密切.培育高类胡萝卜素油菜品种能有效地提升菜籽油的营养保健价值和稳定性,是今后油菜品质育种的一个新方向.研究建立快速、准确和简便的油菜种子类胡萝卜素总量测定方法,是发掘高类胡萝卜素油菜种质资源和开展高类胡萝卜素油菜育种的前提条件.本实验研究了不同提取溶剂、时间、料液比等因素对油菜种子中类胡萝卜素的提取效果和测定结果的影响,确定了最佳的提取和测定条件.建立了快速、准确和简便的油菜种子类胡萝卜素总量测定方法,并测定了30份不同类型油菜种质资源种子中类胡萝卜素总量,为进一步发掘高类胡萝卜素油菜种质资源和开展高类胡萝卜素油菜育种创造了条件.