Comparison of different cultivation modes and light intensities using mono‐cultures and co‐cultures of Haematococcus pluvialis and Chlorella zofingiensis

BACKGROUND: Recent studies indicate that microalgal cultivation using organic carbon sources has the potential to provide high yields. Haematococcus pluvialis and Chlorella zofingiensis, two important carotenoid producers, were selected for co‐culture cultivations to utilize the unique advantages of both organisms. A co‐culture production process was investigated in terms of the effects of organic carbon source, co‐cultivation method, and light intensity on carotenoid production. RESULTS: The addition of 5 g L^?1 glucose resulted in a growth rate of 0.60 day^?1 for H. pluvialis and 0.59 day^?1 for C. zofingiensis, which were higher than those for other carbon sources tested and the control group. Incremental increase of light intensity instead of direct increase to 170 µE m^?2s^? prevented cell loss in both cultures. Co‐cultivation based on cell numbers (60% H. pluvialis and 40% C. zofingiensis) prevented population domination of one microalgae over the other. The biomass production rate of the co‐culture was higher (0.61 g L^?1 day^?1) in glucose‐enriched medium. The total carotenoid content of the co‐culture in the control culture was higher (0.83 mg total carotenoids g^?1 cell) than that obtained in glucose‐enriched medium (0.54 mg total carotenoids g^?1 cell) but not as high as the amounts reached in mono‐cultures. CONCLUSION: Total carotenoid content of the mono‐cultures gave higher yields in standard bold basal medium (BBM). Preliminary high performance liquid chromatography (HPLC) studies indicated a variation in the amounts of astaxanthin isomers produced. Further studies are in progress to determine the effects of carbon‐enriched media and co‐cultivation on the type of isomers and caretenoids produced. Copyright © 2010 Society of Chemical Industry     

Comparison of Extraction Methods for Recovery of Astaxanthin from Haematococcus pluvialis

Solvent extraction, ultrasound assisted extraction (UAE), and microwave assisted extraction (MAE) were examined for the extraction of astaxanthin from Haematococcus pluvialis. In all cases, acetone was found to give the highest astaxanthin recovery compared with other selected solvents, i.e., methanol, ethanol, and acetonitrile. Among the various methods, MAE at 75°C for 5 min resulted in the highest astaxanthin recovery (74 ± 4%).     

Potential of Monoraphidium sp. GK12 for energy‐saving astaxanthin production

BACKGROUND: Astaxanthin is a chemical used in aquaculture, pharmaceuticals, foods, and cosmetics. Although Haematococcus pluvialis has been studied intensively as an astaxanthin producer, its cultivation requires stringent contamination control and nutrition levels, thus rendering astaxanthin production energy‐consuming. In the present study, the medium composition for cultivation of Monoraphidium sp. GK12, a newly isolated astaxanthin‐producing microalga, was improved and photoautotrophic cultivation was attempted to examine its potential for application to energy‐saving astaxanthin production. RESULTS: A cost‐saving medium composed only of mineral salts was found to be suitable for photoautotrophic GK12 cultivation. A specific growth rate of 0.57 day^?1 was obtained, which is higher than that of H. pluvialis. A high cell density of around 2.0 × 10⁷ cells mL^?1 was accomplished in outdoor batch cultivation, while outgrowth of contaminants was not observed. GK12 was also cultivated in a continuous photobioreactor, and the introduction of 1% CO₂ stimulated GK12 growth and astaxanthin productivity. CONCLUSION: Control of temperature, illumination, and culture pH is unnecessary for GK12 outdoor cultivation and no outgrowth of contaminants was observed, suggesting an advantage over H. pluvialis outdoor cultivation. Besides improving Haematococcus cultivation methods, research on GK12 as an alternative astaxanthin‐producer may contribute to the establishment of energy‐saving astaxanthin production. Copyright © 2008 Society of Chemical Industry     

转光膜在雨生红球藻养殖上的应用

通过对雨生红球藻在不同光质条件下生长的比较,确定了红色光有利于藻生长,进而用2.5 L气升式光照反应器在转光膜及普通PE膜下培养藻进行对比,结果显示雨生红球藻生物量、色素、光合活性等几项生物指标在转光膜条件下明显高于普通PE膜. 在气升式反应器内培养的藻细胞,接种9 d,虾青素含量可达3.57 mg/L,叶绿素浓度达到12.42 mg/L,干重提高8.8%以上.     

Optimization of Microwave-Assisted Extraction of Astaxanthin from Haematococcus Pluvialis by Response Surface Methodology and Antioxidant Activities of the Extracts

Abstract

Microwave-assisted extraction (MAE) was applied for the extraction of astaxanthin from Haematococcus pluvialis and response surface methodology (RSM) was used to optimize extraction parameters to the content of astaxanthin. Four independent variables such as microwave power (W), extraction time (sec), solvent volume (mL), and the number of extraction were optimized in this paper. The optimal conditions were determined and tri-dimensional response surfaces were plotted from the mathematical models. The F-test and p-value indicated that microwave power, extraction time, the number of extraction, and their quadratic had a highly significant effect on the response value (p <0.01), then the solvent volume and the interaction effects of microwave power and the number of extraction also displayed significant effect (p <0.05). Considering the extraction efficiency, the optimized conditions of MAE were as follows: microwave power was 141 W, extraction time 83 sec, solvent volume 9.8 mL, the number of extraction four times. About 594 ± 3.02 µg astaxanthin was extracted from Haematococcus pluvialis the dried powders (100 mg) under the optimal conditions, and it close to the predicted contents (592 µg). The antioxidant activities of the extracts obtained under optimal conditions were analyzed, and the results showed that the extracts presented strong ability of inhibiting the peroxidantion of linoleic acid, exhibited strong radical-scavenging properties against the DPPH, as well as strong reducing power.     

Adaptive Responses of Citrus grandis Leaves to Copper Toxicity Revealed by RNA-Seq and Physiology

Copper (Cu)-toxic effects on Citrus grandis growth and Cu uptake, as well as gene expression and physiological parameters in leaves were investigated. Using RNA-Seq, 715 upregulated and 573 downregulated genes were identified in leaves of C. grandis seedlings exposed to Cu-toxicity (LCGSEC). Cu-toxicity altered the expression of 52 genes related to cell wall metabolism, thus impairing cell wall metabolism and lowering leaf growth. Cu-toxicity downregulated the expression of photosynthetic electron transport-related genes, thus reducing CO₂ assimilation. Some genes involved in thermal energy dissipation, photorespiration, reactive oxygen species scavenging and cell redox homeostasis and some antioxidants (reduced glutathione, phytochelatins, metallothioneins, l-tryptophan and total phenolics) were upregulated in LCGSEC, but they could not protect LCGSEC from oxidative damage. Several adaptive responses might occur in LCGSEC. LCGSEC displayed both enhanced capacities to maintain homeostasis of Cu via reducing Cu uptake by leaves and preventing release of vacuolar Cu into the cytoplasm, and to improve internal detoxification of Cu by accumulating Cu chelators (lignin, reduced glutathione, phytochelatins, metallothioneins, l-tryptophan and total phenolics). The capacities to maintain both energy homeostasis and Ca homeostasis might be upregulated in LCGSEC. Cu-toxicity increased abscisates (auxins) level, thus stimulating stomatal closure and lowering water loss (enhancing water use efficiency and photosynthesis).     

Astaxanthin extraction from Haematococcus pluvialis using CO2-expanded ethanol

Microalgae represent diverse branch of microorganism that can produce a wide range of unique functional ingredients that can be used in food, cosmetics, pharmaceuticals, and energy. Among them, Haematococcus pluvialis is known for accumulating the highest levels of a potent natural antioxidant, astaxanthin, which has demonstrated positive health effects. Therefore, the aim of numerous studies has been to develop novel and efficient extraction techniques to produce high-quality (purity and antioxidant activity) extracts, while complying with the Green Chemistry Principles. Supercritical CO₂ (scCO₂) emerges as an alternative to organic solvents because of its high selectivity and bioactivity-preserving qualities. Nevertheless, astaxanthin is a large molecule with low solubility in scCO₂ that usually requires long extractions at high pressures. Ethanol has been used as co-solvent to increase astaxanthin solubility in scCO₂. In this work, a Box–Behnken experimental design was used to study the effects of operating pressure (20–35 MPa), temperature (40–70 °C), and ethanol content in scCO₂ (0–13%, w/w) on the yield, astaxanthin content, and antioxidant activity of H. pluvialis extract. Results showed that ethanol content in CO₂ has a more significant effect on all responses than pressure and temperature. These results lead us to investigate the effect of a further increase in ethanol content, up to the region of gas-expanded liquids. We studied the effects of temperature (30–60 °C) and ethanol content (50–70%, w/w) at a fixed pressure (7 MPa) on the same response variables using CO₂-expanded ethanol (CXE). Results showed that temperature and ethanol content had a significant influence on astaxanthin yield and antioxidant activity. Also, the overall responses of CXE surpassed scCO₂ extractions to match conventional extraction with acetone, maintaining high quality extracts, thus validating the use of this new type of green technology for extraction of high-value compounds.     

Inverse and Concordant Mucosal Pathway Gene Expressions in Inflamed and Non-Inflamed Ulcerative Colitis Patients: Potential Relevance to Aetiology and Pathogenesis

Ulcerative colitis (UC) arises from a complex interplay between host and environmental factors, but with a largely unsolved pathophysiology. The pathophysiology was outlined by RNA-sequencing of mucosal biopsies from non-inflamed and inflamed colon of UC patients (14 and 17, respectively), and from 27 patients without intestinal inflammation. Genes differentially expressed (DE), or present in enriched gene sets, were investigated using statistical text analysis of functional protein information. Compared with controls, inflamed and non-inflamed UC mucosa displayed 9360 and 52 DE genes, respectively. Seventy-three non-pseudogenes were DE relative to both gender and inflammation. Mitochondrial processes were downregulated in inflamed and upregulated in non-inflamed UC mucosa, whereas angiogenesis and endoplasmic reticulum (ER) stress were upregulated in both tissue states. Immune responses were upregulated in inflamed mucosa, whereas the non-inflamed UC mucosa presented both up- and downregulated gene sets. DE and enriched genes overlapped with genes present in inflammatory bowel disease genome-wide associated loci (p = 1.43 × 10⁻¹⁸), especially regarding immune responses, respiratory chain, angiogenesis, ER stress, and steroid hormone metabolism. Apart from confirming established pathophysiological mechanisms of immune cells, our study provides evidence for involvement of less described pathways (e.g., respiratory chain, ER stress, fatty-acid oxidation, steroid hormone metabolism and angiogenesis).