Determination of astaxanthin in Haematococcus pluvialis by first-order derivative spectrophotometry

A highly selective, convenient, and precise method, first-order derivative spectrophotometry, was applied for the determination of astaxanthin in Haematococcus pluvialis. Ethyl acetate and ethanol (1:1, v/v) were found to be the best extraction solvent tested due to their high efficiency and low toxicity compared with nine other organic solvents. Astaxanthin coexisting with chlorophyll and beta-carotene was analyzed by first-order derivative spectrophotometry in order to optimize the conditions for the determination of astaxanthin. The results show that when detected at 432 nm, the interfering substances could be eliminated. The dynamic linear range was 2.0-8.0 microg/mL, with a correlation coefficient of 0.9916. The detection threshold was 0.41 microg/mL. The RSD for the determination of astaxanthin was in the range of 0.01-0.06%; the results of recovery test were 98.1-108.0%. The statistical analysis between first-order derivative spectrophotometry and HPLC by T-testing did not exceed their critical values, revealing no significant differences between these two methods. It was proved that first-order derivative spectrophotometry is a rapid and convenient method for the determination of astaxanthin in H. pluvialis that can eliminate the negative effect resulting from the coexistence of astaxanthin with chlorophyll and beta-carotene.     

Comparison of Different Methods for Extracting the Astaxanthin from Haematococcus pluvialis: Chemical Composition and Biological Activity

In this study, the impact of different cell disruption techniques (high-pressure micro fluidization (HPMF), ionic liquids (ILs), multi-enzyme (ME), and hydrochloric acid (HCl)) on the chemical composition and biological activity of astaxanthin (AST) obtained from Haematococcus pluvialis was investigated. Results indicated that all cell disruption techniques had a significant effect on AST composition, which were confirmed by TLC and UPC² analysis. AST recovery from HCl (HCl-AST) and ILs (ILs-AST) cell disruption techniques was dominant by free and monoesters AST, while AST recovery from HPMF (HPMF-AST) and ME (ME-AST) cell disruption techniques was composed of monoesters, diesters, and free AST. Further biological activity analysis displayed that HCl-AST showed the highest ABTS and DPPH activity, while ILs-AST showed better results against the ORAC assay. Additionally, ILs-AST exhibits a stronger anti-proliferation of HepG2 cells in a dose-dependent manner, which was ascribed to AST-induced ROS in to inhibit the proliferative of cancer cells.     

Haematococcus pluvialis as a Potential Source of Astaxanthin with Diverse Applications in Industrial Sectors: Current Research and Future Directions

Haematococcus pluvialis, a green microalga, appears to be a rich source of valuable bioactive compounds, such as astaxanthin, carotenoids, proteins, lutein, and fatty acids (FAs). Astaxanthin has a variety of health benefits and is used in the nutraceutical and pharmaceutical industries. Astaxanthin, for example, preserves the redox state and functional integrity of mitochondria and shows advantages despite a low dietary intake. Because of its antioxidant capacity, astaxanthin has recently piqued the interest of researchers due to its potential pharmacological effects, which include anti-diabetic, anti-inflammatory, and antioxidant activities, as well as neuro-, cardiovascular-, ocular, and skin-protective properties. Astaxanthin is a popular nutritional ingredient and a significant component in animal and aquaculture feed. Extensive studies over the last two decades have established the mechanism by which persistent oxidative stress leads to chronic inflammation, which then mediates the majority of serious diseases. This mini-review provides an overview of contemporary research that makes use of the astaxanthin pigment. This mini-review provides insight into the potential of H. pluvialis as a potent antioxidant in the industry, as well as the broad range of applications for astaxanthin molecules as a potent antioxidant in the industrial sector.     

Effect of CO2 Flow Rate on the Extraction of Astaxanthin and Fatty Acids from Haematococcus pluvialis Using Supercritical Fluid Technology

Haematococcus pluvialis is the largest producer of natural astaxanthin in the world. Astaxanthin is a bioactive compound used in food, feed, nutraceutics, and cosmetics. In this study, astaxanthin extraction from H. pluvialis by supercritical fluid extraction was evaluated. The effects of temperature (40 and 50 °C), pressure (40 and 50 MPa), and CO₂ flow rate (2 and 4 L/min) were investigated. The results showed that the highest astaxanthin recovery was obtained at 50 °C/50 MPa and the CO₂ flow rates evaluated had no significant effect. It was possible to achieve astaxanthin recoveries of 95% after 175 min for a CO₂ flow rate of 2 L/min, and 95 min for CO₂ flow rate of 4 L/min. The ω-6/ω-3 ratios obtained were similar in all conditions, reaching 0.87, demonstrating that the extracts from H. pluvialis by SFE are rich in unsaturated fatty acids (UFA) which increases their positive effects when used as a functional ingredient in food.     

Investigation of the maximum quantum yield of PS II in Haematococcus pluvialis cell cultures during growth: effects of chemical or high-intensity light treatment

In this study, we investigated the increase in photosynthetic quantum yield that occurs in advance of increased microalgal growth. Haematococcus pluvialis was cultivated under normal conditions; the number of cells, the maximum quantum yield of photosystem II (F(v)/F(m)), and optical density were measured. We observed an increase in F(v)/F(m) approximately 72h prior to the cell growth phase. To confirm the relationship between photosynthetic yield and growth, samples were treated with several chemicals under high-intensity light illumination and control conditions to inhibit photosystem II and induce a decrease in the quantum photosynthetic yield. The samples were exposed to high-intensity light at an irradiance of 400μmol photonsm(-2)s(-1) for varied amount of time and were treated with chemicals such as 3-(3,4-dichlorophenyl)-1,1-dimethylurea, nigericin sodium salt and valinomycin. We observed that both the photooxidation of photosystem II reaction centers and the formation of transmembrane electrochemical gradients led to an initial decrease in fluorescence yield after the onset of high-intensity light illumination. We also observed that treatment of high-intensity light illuminated cells with antibiotics after adaptation to moderate light intensities caused a difference in photosynthetic activity. In conclusion, the maximum quantum yield of photosystem II is obtained prior to the cell growth phase and can therefore be used as a prediction parameter for cell growth.     

Sodiation as a tool for enhancing the diagnostic value of MALDI‐TOF/TOF‐MS spectra of complex astaxanthin ester mixtures from Haematococcus pluvialis

The microalga Haematococcus pluvialis produces the pigment astaxanthin mainly in esterified form with a multitude of fatty acids, which results in a complex mixture of carotenol mono‐ and diesters. For rapid fingerprinting of these esters, matrix‐assisted laser desorption ionization time of flight mass spectrometry (MALDI‐TOF/TOF‐MS) might be an alternative to traditional chromatographic separation combined with MS. Investigation of ionization and fragmentation of astaxanthin mono‐ and diester palmitate standards in MALDI‐TOF/TOF‐MS showed that sodium adduct parent masses [M + Na]⁺ gave much simpler MS² spectra than radical / protonated [M]^+● / [M + H]⁺ parents. [M + Na]⁺ fragments yielded diagnostic polyene‐specific eliminations and fatty acid neutral losses, whereas [M]^+● / [M + H]⁺ fragmentation resulted in a multitude of non‐diagnostic daughters. For diesters, a benzonium fragment, formed by polyene elimination, was required for identification of the second fatty acid attached to the astaxanthin backbone. Parents were forced into [M + Na]⁺ ionization by addition of sodium acetate, and best signal‐to‐noise ratios were obtained in the 0.1 to 1.0 mM range. This method was applied to fingerprinting astaxanthin esters in a crude H. pluvialis extract. Prior to MALDI‐TOF/TOF‐MS, the extract was fractionated by normal phase Flash chromatography to obtain fractions enriched in mono‐ and diesters and to remove pheophytin a, which compromised monoester signals. All 12 types of all‐trans esterified esters found in LC were identified with MALDI‐TOF/TOF‐MS, with the exception of two minor monoesters. Copyright © 2013 John Wiley & Sons, Ltd.     

半连续乳液聚合动力学模型

介绍半连续乳液聚合动力学模型的研究进展及建模方法，归结出３种形式模型，基础模型，具体模型与通用模型，指出通常从一些假定出发，根据质量平衡，能量平衡反应机理建立模型方程，通过模型参数估计，已有模型参数的利用及模型变量的求取，使用数值方法求解模型。     

Studies on the genetic variation of the green unicellular alga Haematococcus pluvialis (Chlorophyceae) obtained from different geographical locations using ISSR and RAPD molecular marker

Haematococcus pluvialis (Flotow) is a unicellular green alga, which is considered to be the best astaxanthin-producing organism. Molecular markers are suitable tools for the purpose of finding out genetic variations in organisms; however there have been no studies conducted on ISSR or RAPD molecular markers for this organism. The DNA of 10 different strains of H. pluvialis (four strains from Iran, two strains from Finland, one strain from Switzerland and three strains from the USA) was extracted. A genetic similarity study was carried out using 14 ISSR and 12 RAPD primers. Moreover, the molecular weights of the bands produced ranged from 0.14 to 3.4 Kb. The PCA and dendrogram clustered the H. pluvialis strains into various groups according to their geographical origin. The lowest genetic similarity was between the Iran2 and USA2 strains (0.08) and the highest genetic similarity was between Finland1 and Finland2 (0.64). The maximum numbers of bands produced by the ISSR and RAPD primers were 35 and 6 bands, respectively. The results showed that ISSR and RAPD markers are useful for genetic diversity studies of Haematococcus as they showed geographical discrimination.     

Mixotrophic Cultivation of Microalgae for Biodiesel Production: Status and Prospects

Biodiesel from microalgae provides a promising alternative for biofuel production. Microalgae can be produced under three major cultivation modes, namely photoautotrophic cultivation, heterotrophic cultivation, and mixotrophic cultivation. Potentials and practices of biodiesel production from microalgae have been demonstrated mostly focusing on photoautotrophic cultivation; mixotrophic cultivation of microalgae for biodiesel production has rarely been reviewed. This paper summarizes the mechanisms and virtues of mixotrophic microalgae cultivation through comparison with other major cultivation modes. Influencing factors of microalgal biodiesel production under mixotrophic cultivation are presented, development of combining microalgal biodiesel production with wastewater treatment is especially reviewed, and bottlenecks and strategies for future commercial production are also identified.     

Semi-continuous Cultivation of <Emphasis Type="Italic">Chlorella vulgaris</Emphasis> for Treating Undigested and Digested Dairy Manures

The present study, based on a previous batch-wise experiment, investigated a lab-scale semi-continuous cultivation of green microalgae Chlorella vulgaris (UTEX 2714), as a useful means for nutrient reduction as well as production of algal biomass which can be used as potential feedstock for the production of biofuel and other commodities, on 20× diluted dairy manures. Both undigested and digested samples were applied in parallel experiments for comparison regarding the requirements of hydraulic retention times (HRTs), removal efficiencies of nitrogen, phosphorus, and chemical oxygen demand (COD), biomass productivities, and CO₂ sequestration abilities. It was demonstrated that algae grown in undigested dairy manure achieved removal rates of 99.7%, 89.5%, 92.0%, and 75.5% for NH₄⁺–N, TN, TP, and COD, respectively, under a 5-day HRT, while the HRT had to extend to 20 days in order to achieve 100.0% removal of NH₄⁺–N in digested one with simultaneous removals of 93.6% of TN, 89.2% of TP, and 55.4% of COD. The higher organic carbon contained in undigested dairy manure helped boost the growth of mixotrophic Chlorella, thus resulting in a much shorter HRT needed for complete removal of NH₄⁺–N. Moreover, algae grown in digested dairy manure provided more penitential than those grown in undigested one in CO₂ sequestration per milligram of harvested dried biomass (1.68 mg CO₂/mg dry weight (DW) vs 0.99 mg CO₂/mg DW), but did not surpass in total the amount of CO₂ sequestered on a 15-day period basis because of the better productivity gained in undigested dairy manure.     

Screening,Growth Medium Optimisation and Heterotrophic Cultivation of Microalgae for Biodiesel Production

This article presents a study on screening of microalgal strains from the Peking University Algae Collection and heterotrophic cultivation for biodiesel production of a selected microalgal strain. Among 89 strains, only five were capable of growing under heterotrophic conditions in liquid cultures and Chlorella sp. PKUAC 102 was found the best for the production of heterotrophic algal biodiesel. Composition of the growth medium was optimised using response surface methodology and optimised growth conditions were successfully used for cultivation of the strain in a fermentor. Conversion of algal lipids to fatty acid methyl esters (FAMEs) showed that the lipid profile of the heterotrophically cultivated Chlorella sp. PKUAC 102 contains fatty acids suitable for biodiesel production.     

3-Chloropropyltrialkoxysilanes—Key Intermediates for the Commercial Production of Organofunctionalized Silanes and Polysiloxanes

Silicon and carbon may be looked upon as the two fundamental antipodes with respect to the naturally occurring forms of matter and life on earth: Silicon plays the key role in the “petrification” of inanimate matter, while carbon is the representative element for the “incarnation” of living organisms. If a scientist were to search for a highly efficient method of uniting inorganic and organic matter, he would, at a very early stage of attempting to solve this problem, combine these two natural principles of matter and think of “bireactive” molecules containing a silicon functionality for bonding to inorganic material on the one hand and a carbon functionality for anchoring to an organic counterpart on the other. The title compounds and their derivatives epitomize such molecules: they are wanderers between both (chemical) worlds. Owing to their bifunctionality they are capable of binding to inorganic (especially siliceous) systems as well as to organic polymers. Whether their commercial application concerns the strengthening of polyester resins with glass fibers for use in boatbuilding or the incorporation of silica as a filler in rubber mixtures for the manufacture of wear-resistant tires or even the immobilization of enzymes on glass spheres for carrying out reactions in enzyme reactors—in all cases organofunctionalized silanes guarantee a reliable and permanent union between two otherwise “incompatible” material systems.     

Microbial Pretreatment of Corn Stovers by Solid-State Cultivation of Phanerochaete chrysosporium for Biogas Production

The microbial pretreatment of corn stover and corn stover silage was achieved via the solid-state cultivation of Phanerochaete chrysosporium; pretreatment effects on the biodegradability and subsequent anaerobic production of biogas were investigated. The peak levels of daily biogas production and CH₄ yield from corn stover silage were approximately twice that of corn stover. Results suggested that ensiling was a potential pretreatment method to stimulate biogas production from corn stover. Surface morphology and Fourier-transform infrared spectroscopy analyses demonstrated that the microbial pretreatment of corn stover silage improved biogas production by 10.5 to 19.7 % and CH₄ yield by 11.7 to 21.2 % because pretreatment could decrease dry mass loss (14.2 %) and increase substrate biodegradability (19.9 % cellulose, 32.4 % hemicellulose, and 22.6 % lignin). By contrast, the higher dry mass loss in corn stover (55.3 %) after microbial pretreatment was accompanied by 54.7 % cellulose, 64.0 % hemicellulose, and 61.1 % lignin degradation but did not significantly influence biogas production.     

Simple Strategy of Repeated Batch Cultivation for Enhanced Production of 1,3-Propanediol Using Clostridium diolis

Repeated batch cultivation (empty-and-fill protocol) using obligate anaerobe Clostridium diolis was attempted in the present study to improve the production of 1,3-propanediol (1,3-PD). In repeated batch operation, 20?% (v/v) culture broth was removed from the bioreactor and supplemented with an equal volume of fresh nutrient medium when the residual glycerol concentration in the bioreactor decreased below 15?g/L. Four cycles of culture broth withdrawal and subsequent replacement resulted in achieving a 1,3-PD concentration of 67.8?g/L with a productivity of 1.04?g/L/h at the end of 65?h. This represented a 1,3-PD concentration and productivity enhancement by 2.6-fold and 1.5-fold, respectively, as compared to batch 1,3-PD fermentation. This is the first report on the use of repeated batch mode of bioreactor operation for enhanced 1,3-PD production.     

Development of Novel Molecular Catalysts “Phosphazene Catalysts” for Commercial Production of Highly Advanced Polypropylene Glycols

This contribution reports development and an application of high-performance catalysts, phosphazene catalysts (PZN and PZO) with high reactivity and selectivity for ring-opening polymerization reactions of propylene oxides that provide highly advanced polypropylene glycols. The produced polypropylene glycols with high purity and narrow molecular weight distributions have been commercialized as raw materials for environmentally- and people-friendly polyurethane foams (for car sheet cushions).     

Mass Cultivation of Microalgae on Animal Wastewater: a Sequential Two-Stage Cultivation Process for Energy Crop and Omega-3-Rich Animal Feed Production

In this study, 97 microalgal strains purchased from algae bank and 50 microalgal strains isolated from local waters in Minnesota were screened for their adaptability growing on a 20-fold diluted digested swine manure wastewater (DSMW). A pool of candidate strains well adapted to the DSMW was established through a high-throughput screening process. Two top-performing facultative heterotrophic strains with high growth rate (0.536?day^?1 for UMN 271 and 0.433?day^?1 for UMN 231) and one strain with high omega-3 unsaturated fatty acid (EPA, 3.75?% of total fatty acids for UMN 231) were selected. Subsequently, a sequential two-stage mixo-photoautotrophic culture strategy was developed for biofuel and animal feed production as well as simultaneous swine wastewater treatment using above two strains. The maximal biomass concentration and lipid content at the first and second stages reached 2.03?g/L and 23.0?%, and 0.83?g/L and 19.0?% for UMN 271 and UMN 231, respectively. The maximal nutrient removals for total phosphorus and ammonia after second-stage cultivation were 100 and 89.46?%, respectively. The experiments showed that this sequential two-stage cultivation process has great potential for economically viable and environmentally friendly production of both renewable biofuel and high-value animal feed and at the same time for animal wastewater treatment.     

Investigation on the Cultivation Conditions of a Newly Isolated Fusarium Fungal Strain for Enhanced Lipid Production

Applied Biochemistry and Biotechnology - Fusarium equiseti UMN-1 fungal strain isolated from soybean is selected as a potential oleaginous fungal strain for biodiesel generation. It possesses...     

Semi-continuous High Speed Gas Analysis of Generated Vapors of Chemical Warfare Agents

A method is presented for the continuous analysis of generated vapors of the nerve agents soman and sarin and the blistering agent sulfur mustard. By using a gas sampling valve and a very short (15 cm) column connected to an on-column injector with a “standard length” column, the system can either be calibrated or used for performing high speed gas analyses. When using a flame ionization detector, the detection limit was ca. 100 ppb (ca. 0.5–1.0 mg/m³). This technique is applied in inhalation toxicokinetic studies of nerve agents and mustard gas in the guinea pig.     

Semi-continuous Heterophase Polymerization of n-Butyl Methacrylate: Effect of Monomer Feeding Rate

The effect of monomer feeding rate on particle size, molar masses, glass transition and tacticity of poly(n-butyl methacrylate) (PBMA) nanoparticles synthesized by semi-continuous heterophase polymerization under monomer-starved conditions is reported. Three feed rates were examined. Highly monomer-starved conditions at the two slower addition rates were confirmed by the low amount of residual monomer throughout the reaction and by the fact that the instantaneous polymerization and feeding rates became similar at later stages of the reaction. Under these conditions, polymer particles in the nanometer range (30 to 35 nm) were obtained. Glass transition temperatures are substantially higher than those reported for commercial PBMA. Polymers tacticity was determined by ¹³C-NMR spectroscopy. NMR measurements confirm that the syndiotactic content of the PBMA synthesized here is larger than those of the commercial ones made by free-radical polymerization. Molar masses are much lower than those expected from termination by chain transfer to monomer, which is the typical termination mechanism in microemulsion polymerization.     

Strategies to Overcome Oxygen Transfer Limitations During Hairy Root Cultivation of Azadiracta indica for Enhanced Azadirachtin Production

The vast untapped potential of hairy root cultures as a stable source of biologically active chemicals has focused the attention of scientific community toward its commercial exploitation. However, the major bottleneck remains its successful scale-up. Due to branching, the roots form an interlocked matrix that exhibits resistance to oxygen transfer. Thus, present work was undertaken to develop cultivation strategies like optimization of inlet gas composition (in terms of % (v/v) O₂ in air), air-flow rate and addition of oxygen vectors in the medium, to curb the oxygen transfer limitations during hairy root cultivation of Azadirachta indica for in vitro azadirachtin (a biopesticide) production. It was found that increasing the oxygen fraction in the inlet air (in the range, 20–100% (v/v) O₂ in air) increased the azadirachtin productivity by approximately threefold, to a maximum of 4.42 mg/L per day (at 100% (v/v) O₂ in air) with respect to 1.68 mg/L per day in control (air with no oxygen supplementation). Similarly, increasing the air-flow rate (in the range, 0.3–2 vvm) also increased the azadirachtin productivity to a maximum of 1.84 mg/L per day at 0.8 vvm of air-flow rate. On the contrary, addition of oxygen vectors (in the range, 1–4% (v/v); hydrogen peroxide, toluene, Tween 80, kerosene, silicone oil, and n-hexadecane), decreased the azadirachtin productivity with respect to control (1.76 mg/L per day).