Ketocarotenoid production by a mutant of Chlorococcum sp. in an outdoor tubular photobioreactor

A mutant, MA-1, of Chlorococcum sp., grown in batch culture, produced about 54 mg ketocarotenoids/l with 10 mM nitrogen. The accumulation rate of these ketocarotenoids was independent of the nitrogen concentration under sunlight illumination. Fed-batch cultures showed poor growth and the average productivity of ketocarotenoids dropped from 2.6 mg/l day to 1.6 mg/l day in the two consecutive fed-batch runs.     

Comparative analysis of astaxanthin and its esters in the mutant E1 of Haematococcus pluvialis and other green algae by HPLC with a C30 column

A gradient reversed-phase high-performance liquid chromatography (HPLC) method using a C30 col-umn was developed for the simultaneous determination of astaxanthin, astaxanthin monoesters and astaxanthin diesters in the green algae Chlorococcum sp., Chlorella zofingiensis, Haematococcus plu-vialis and the mutant E1, which was obtained from the mutagenesis of H. pluvialis by exposure to UV-irradiation and ethyl methanesulphonate (EMS) with subsequent screening using nicotine. The re-sults showed that the contents of total astaxanthins including free astaxanthin and astaxanthin esters ranged from 1.4 to 30.9 mg/g dry biomass in these green algae. The lower total astaxanthin levels (< 2 mg/g dry biomass) were detected in the green algae Chlorococcum sp. and C. zofingiensis. The higher total astaxanthin levels (>16 mg/g dry biomass) were found in the green alga H. pluvialis and its mutant E1. It is notable that the mutant E1 is found to have considerably higher amounts of total astaxanthin (30.9 mg/g) as compared to the wild strain of H. pluvialis (16.1 mg/g). This indicates that UV-irradiation and EMS compound mutagenesis with subsequent screening using nicotine is an effective method for breeding of a high-producing astaxanthin strain of H. pluvialis. In addition, the green alga C. zofingien-sis had a remarkably higher percentage of astaxanthin diesters (76.3% of total astaxanthins) and a re-markably lower percentage of astaxanthin monoesters (18.0% of total astaxanthins) in comparison with H. pluvialis (35.5% for diesters and 60.9% for monoesters), the mutant E1 (49.1% and 48.1%) and Chlorococcum sp. (18.0% and 58.6%).     

Carotenoid content of chlorophycean microalgae: factors determining lutein accumulation in Muriellopsis sp. (Chlorophyta)

Fifteen strains of chlorophycean microalgae have been investigated with regard to their carotenoid profile. Lutein, beta-carotene and violaxanthin were present in virtually all of the strains, lutein, in general, being the most abundant carotenoid, whereas canthaxanthin and astaxanthin were found in some strains only. Chlorella fusca SAG 211-8b, Chlorococcum citriforme SAG 62.80, Muriellopsis sp., Neospongiococcum gelatinosum SAG B 64.80 and Chlorella zofingiensis CCAP 211/14 exhibited high lutein levels, the latter strain containing in addition substantial amounts of astaxanthin. Muriellopsis sp. was further characterized, since besides a high lutein content (up to 35 mg l(-1) culture), it had the highest growth rate (up to 0.17-0.23 h(-1)) and maximal standing cell density (up to 8 x 10(10) cells l(-1) culture). These levels of lutein are in the range of those reported for astaxanthin in Haematococcus and for beta-carotene in Dunaliella, microalgae of recognized interest for the production of these carotenoids. Lutein content of Muriellopsis sp. increased during the exponential phase of growth, with the highest value being recorded in the early stationary phase. Maximum levels of lutein in Muriellopsis sp. cultures were recorded at 20-40 mM NaNO3, 2-100 mM NaCl, 460 micromol photon m(-2) s(-1), pH 6.5 and 28 degrees C, conditions which were, in general, also optimal for cell growth. Growth-limiting conditions, such as pH values of 6 or 9 and a temperature of 33 degrees C, were found to stimulate carotenogenesis in Muriellopsis sp. This strain represents a potential source of lutein, a commercially interesting carotenoid of application in aquaculture and poultry farming, as well as in the prevention of cancer and diseases related to retinal degeneration.     

Enhanced accumulation of secondary carotenoids in a mutant of the green alga, Chlorococcum sp.

A colour mutant of the unicellular green alga Chlorococcum sp. was obtained by visual colour detection method on plates with medium containing sodium azide. The growth of the mutant MA-1 was more susceptible to azide compared with the wild type, whereas the total secondary carotenoid (SC)synthesis was more resistant to the inhibitor. The azide concentration that inhibited SC formation by 50% (I₅₀) was ten times higher than that required for the wild type. The mutant was stable over several consecutive subculturings in the absence of azide. The indoor and outdoor studies showed that the mutant could synthesise more than 2-fold of the total SC and astaxanthin of the wild type. The mutant MA-1 could be a natural source of SC and astaxanthin. This revised version was published online in August 2006 with corrections to the Cover Date.     

Composition and accumulation of secondary carotenoids in Chlorococcum sp.

A locally isolated Chlorococcum sp. could accumulate astaxanthin and its esters as secondary carotenoids. The secondary carotenoids could reach a concentration of 5.2 mg g⁻¹ d. wt, and were located in the cytoplasm and chloroplast as globules. Cells grew best at pH 8.0 and 30 °C, at which the growth rate was about 0.066 h⁻¹. Acidic condition (pH 5.5 and 6.5) and slightly elevated temperature (35 °C) enhanced the cellular accumulation of astaxanthin. Outdoor studies indicated that Chlorococcum sp. grew well in a tubular photobioreactor. In medium containing 2 mM and 10 mM NH₄CI, the cellular contents of total secondary carotenoids and astaxanthin reached similar levels (5.0 mg g⁻¹ d. wt and 2.0 mg g⁻¹ d. wt, respectively) in the 15 days of cultivation, while the yield of total secondary carotenoids and astaxanthin in 10 mM NH₄CI were higher (45 mg L⁻¹ and 18 mg L⁻¹, respectively). The advantages of tolerance to high temperature and extreme pH values, relative fast growth rate and ease of cultivation in outdoor system suggest that Chlorococcum sp. could be a potential candidate for mass production of secondary carotenoids in particular astaxanthin. This revised version was published online in September 2006 with corrections to the Cover Date.     

Construction of transplastomic lettuce (Lactuca sativa) dominantly producing astaxanthin fatty acid esters and detailed chemical analysis of generated carotenoids

The plastid genome of lettuce (Lactuca sativa L.) cv. Berkeley was site-specifically modified with the addition of three transgenes, which encoded β,β-carotenoid 3,3′-hydroxylase (CrtZ) and β,β-carotenoid 4,4′-ketolase (4,4′-oxygenase; CrtW) from a marine bacterium Brevundimonas sp. strain SD212, and isopentenyl diphosphate isomerase from a marine bacterium Paracoccus sp. strain N81106. Constructed transplastomic lettuce plants were able to grow on soil at a growth rate similar to that of non-transformed lettuce cv. Berkeley and generate flowers and seeds. The germination ratio of the lettuce transformants (T₀) (98.8 %) was higher than that of non-transformed lettuce (93.1 %). The transplastomic lettuce (T₁) leaves produced the astaxanthin fatty acid (myristate or palmitate) diester (49.2 % of total carotenoids), astaxanthin monoester (18.2 %), and the free forms of astaxanthin (10.0 %) and the other ketocarotenoids (17.5 %), which indicated that artificial ketocarotenoids corresponded to 94.9 % of total carotenoids (230 μg/g fresh weight). Native carotenoids were there lactucaxanthin (3.8 %) and lutein (1.3 %) only. This is the first report to structurally identify the astaxanthin esters biosynthesized in transgenic or transplastomic plants producing astaxanthin. The singlet oxygen-quenching activity of the total carotenoids extracted from the transplastomic leaves was similar to that of astaxanthin (mostly esterified) from the green algae Haematococcus pluvialis.     

Functional characterization of various algal carotenoid ketolases reveals that ketolating zeaxanthin efficiently is essential for high production of astaxanthin in transgenic Arabidopsis

Extending the carotenoid pathway to astaxanthin in plants is of scientific and industrial interest. However, expression of a microbial β-carotene ketolase (BKT) that catalyses the formation of ketocarotenoids in transgenic plants typically results in low levels of astaxanthin. The low efficiency of BKTs in ketolating zeaxanthin to astaxanthin is proposed to be the major limitation for astaxanthin accumulation in engineered plants. To verify this hypothesis, several algal BKTs were functionally characterized using an Escherichia coli system and three BKTs were identified, with high (up to 85%), moderate (～38%), and low (～1%) conversion rate from zeaxanthin to astaxanthin from Chlamydomonas reinhardtii (CrBKT), Chlorella zofingiensis (CzBKT), and Haematococcus pluvialis (HpBKT3), respectively. Transgenic Arabidopsis thaliana expressing the CrBKT developed orange leaves which accumulated astaxanthin up to 2 mg g(-1) dry weight with a 1.8-fold increase in total carotenoids. In contrast, the expression of CzBKT resulted in much lower astaxanthin content (0.24 mg g(-1) dry weight), whereas HpBKT3 was unable to mediate synthesis of astaxanthin in A. thaliana. The none-native astaxanthin was found mostly in a free form integrated into the light-harvesting complexes of photosystem II in young leaves but in esterified forms in senescent leaves. The alteration of carotenoids did not affect chlorophyll content, plant growth, or development significantly. The astaxanthin-producing plants were more tolerant to high light as shown by reduced lipid peroxidation. This study advances a decisive step towards the utilization of plants for the production of high-value astaxanthin.     

Two-stage cultures for the production of astaxanthin from Haematococcus pluvialis

A two-stage culture system was established for the production of astaxanthin from Haematococcus pluvialis. In a first stage green vegetative cells were produced in semicontinuous cultures maintained with daily renewal rates between 10 and 40%. The steady-state cell density decreased with increasing renewal rates. Highest cell productivity, 64 x 10(6) cells l(-1) day(-1) was obtained with a daily renewal rate of 20%. In a second stage the harvested cultures were submitted to high light (240 micromol photon m(-2) s(-1)) under batch conditions for 15 days in order to stimulate the transition to the aplanospore stage and the accumulation of astaxanthin. No decrease in cell density was recorded during the induction period in any of the cultures. Cultures obtained at high renewal rates continued growing during the induction period and no astaxanthin was accumulated until all nitrogen in the media had been consumed. The final concentration of astaxanthin was inversely correlated to the growth rate at which first-stage cultures were maintained. Optimal renewal rate for maximal astaxanthin production depended on the duration of the induction period. After a 12-day induction period the highest astaxanthin production, 5.8 mg l(-1) of semi-continuous culture day -1, was obtained with cultures maintained at a renewal rate of 20%. When the induction period was increased to 15 days maximal astaxanthin productivity, 9.6 mg l(-1) of semi-continuous culture day -1, was obtained from cultures maintained at a renewal rate of 40% despite the much lower astaxanthin concentration achieved in these cultures. Results demonstrate the feasibility of semi-continuous cultivation of H. pluvialis for the two-stage production of astaxanthin.     

Production of fructosyltransferase by <Emphasis Type="Italic">Aureobasidium</Emphasis> sp. ATCC 20524 in batch and two-step batch cultures

A comparison of fructosyltransferase (EC 2.4.1.9) production by Aureobasidium sp. ATCC 20524 in batch and two step batch cultures was investigated in a 1-l stirred tank reactor using a sucrose supply of 200 g/l. Results showed that the innovative cultivation in two step of Aureobasidium sp. produced more fructosyltransferase (FFase) than the single batch culture at the same sucrose concentration with a maximal enzyme production of 523 U/ml, which was 80.5% higher than the one obtained in the batch culture. The production of fructooligosaccharides (FOSs) was also analyzed; their concentration reached a maximum value of 160 g/l the first day in the two-step culture and 127 g/l in the single-batch mode. The use of the two-step batch culture with Aureobasidium sp. ATCC 20524 in allowing the microorganism to grow up prior to the induction of sucrose (second step), proved to be a powerful method for producing fructosyltransferase and FOSs.     

Ketocarotenoid formation in transgenic potato

Potato has been genetically engineered for the production of commercially important ketocarotenoids including astaxanthin (3,3'-dihydroxy 4,4'-diketo-beta-carotene). To support the formation of 3-hydroxylated and 4-ketolated beta-carotene, a transgenic potato line accumulating zeaxanthin due to inactivated zeaxanthin epoxidase was co-transformed with the crtO beta-carotene ketolase gene from the cyanobacterium Synechocystis under a constitutive promoter. Plants were generated which exhibited expression of this gene, resulting in an accumulation of echinenone, 3'-hydroxyechinenone, and 4-ketozeaxanthin in leaves, as well as 3'-hydroxyechinenone, 4-ketozeaxanthin together with astaxanthin in the tuber. The amount of ketocarotenoids formed represent approximately 10-12% of total carotenoids in leaves and tubers. Negative effects on photosynthesis due to the presence of the ketocarotenoids in leaves could be excluded by the determination of variable fluorescence.     

<Emphasis Type="Italic">Nicotiana glauca</Emphasis> engineered for the production of ketocarotenoids in flowers and leaves by expressing the cyanobacterial <Emphasis Type="Italic">crtO</Emphasis> ketolase gene

Nicotiana glauca is a tobacco species that forms flowers with carotenoid-pigmented petals, sepals, pistil, ovary and nectary tissue. The carotenoids produced are lutein, ss-carotene as well as some violaxanthin and antheraxanthin. This tobacco species was genetically modified for ketocarotenoid biosynthesis by transformation with a cyanobacterial crtO ketolase gene under the 35S CaMV promoter. In the transformants, ketocarotenoids were detected in both leaves and flowers. Although astaxanthin was not detected other ketocarotenoids such as 4'-ketolutein, echinenone, 3'-hydroxyechinenone and 4-ketozeaxanthin were present. Accumulation of ketocarotenoids in leaves decreased their photosynthetic efficiency moderately. Under the green house conditions used no impairment of growth and development compared to the wild type was observed. In the crtO-transformants, an unexpected up-regulation of total carotenoid biosynthesis in leaves and especially in flower petals was observed. This led to a total ketocarotenoid concentration in leaves of 136.6 (young) or 156.1 (older) mug/g dry weight and in petals of 165 mug/g dry weight. In our engineered plants, the ketocarotenoid pathway is one step short of astaxanthin. Strategies are discussed to improve N. glauca flowers as a biological system for astaxanthin.     

Ketocarotenoid Production in Soybean Seeds through Metabolic Engineering

The pink or red ketocarotenoids, canthaxanthin and astaxanthin, are used as feed additives in the poultry and aquaculture industries as a source of egg yolk and flesh pigmentation, as farmed animals do not have access to the carotenoid sources of their wild counterparts. Because soybean is already an important component in animal feed, production of these carotenoids in soybean could be a cost-effective means of delivery. In order to characterize the ability of soybean seed to produce carotenoids, soybean cv. Jack was transformed with the crtB gene from Pantoea ananatis, which codes for phytoene synthase, an enzyme which catalyzes the first committed step in the carotenoid pathway. The crtB gene was engineered together in combinations with ketolase genes (crtW from Brevundimonas sp. strain SD212 and bkt1 from Haematococcus pluvialis) to produce ketocarotenoids; all genes were placed under the control of seed-specific promoters. HPLC results showed that canthaxanthin is present in the transgenic seeds at levels up to 52 μg/g dry weight. Transgenic seeds also accumulated other compounds in the carotenoid pathway, such as astaxanthin, lutein, β-carotene, phytoene, α-carotene, lycopene, and β-cryptoxanthin, whereas lutein was the only one of these detected in non-transgenic seeds. The accumulation of astaxanthin, which requires a β-carotene hydroxylase in addition to a β-carotene ketolase, in the transgenic seeds suggests that an endogenous soybean enzyme is able to work in combination with the ketolase transgene. Soybean seeds that accumulate ketocarotenoids could potentially be used in animal feed to reduce or eliminate the need for the costly addition of these compounds.     

Comparative analysis of astaxanthin and its esters in the mutant E1 of Haematococcus pluvialis and other green algae by HPLC with a C30 column

A gradient reversed-phase high-performance liquid chromatography (HPLC) method using a C30 column was developed for the simultaneous determination of astaxanthin, astaxanthin monoesters and astaxanthin diesters in the green algae Chlorococcum sp., Chlorella zofingiensis, Haematococcus pluvialis and the mutant E1, which was obtained from the mutagenesis of H. pluvialis by exposure to UV-irradiation and ethyl methanesulphonate (EMS) with subsequent screening using nicotine. The results showed that the contents of total astaxanthins including free astaxanthin and astaxanthin esters ranged from 1.4 to 30.9 mg/g dry biomass in these green algae. The lower total astaxanthin levels (< 2 mg/g dry biomass) were detected in the green algae Chlorococcum sp. and C. zofingiensis. The higher total astaxanthin levels (>16 mg/g dry biomass) were found in the green alga H. pluvialis and its mutant E1. It is notable that the mutant E1 is found to have considerably higher amounts of total astaxanthin (30.9 mg/g) as compared to the wild strain of H. pluvialis (16.1 mg/g). This indicates that UV-irradiation and EMS compound mutagenesis with subsequent screening using nicotine is an effective method for breeding of a high-producing astaxanthin strain of H. pluvialis. In addition, the green alga C. zofingiensis had a remarkably higher percentage of astaxanthin diesters (76.3% of total astaxanthins) and a remarkably lower percentage of astaxanthin monoesters (18.0% of total astaxanthins) in comparison with H. pluvialis (35.5% for diesters and 60.9% for monoesters), the mutant E1 (49.1% and 48.1%) and Chlorococcum sp. (18.0% and 58.6%). Supported by the Frontier Research Grant of the SCSIO, the Hundred Talents program of Chinese Academy of Sciences, and National Natural Sciences of China projects (Grant No. 40776087)     

Secondary ketocarotenoid astaxanthin biosynthesis in algae: a multifunctional response to stress

Under stressful environments, many green algae such as Haematococcus pluvialis accumulate secondary ketocarotenoids such as canthaxanthin and astaxanthin. The carotenogenesis, responsible for natural phenomena such as red snows, generally accompanies larger metabolic changes as well as morphological modifications, i.e., the conversion of the green flagellated macrozoids into large red cysts. Astaxanthin accumulation constitutes a convenient way to store energy and carbon, which will be used for further synthesis under less stressful conditions. Besides this, the presence of high amount of astaxanthin enhances the cell resistance to oxidative stress generated by unfavorable environmental conditions including excess light, UV-B irradiation, and nutrition stress and, therefore, confers a higher survival capacity to the cells. This better resistance results from the quenching of oxygen atoms for the synthesis itself as well as from the antioxidant properties of the astaxanthin molecules. Therefore, astaxanthin synthesis corresponds to a multifunctional response to stress. In this contribution, the various biochemical, genetic, and molecular data related to the biosynthesis of ketocarotenoids by Haematococcus pluvialis and other taxa are reviewed and compared. A tentative regulatory model of the biochemical network driving astaxanthin production is proposed.     

The Effect of Salt Stress on the Production of Canthaxanthin and Astaxanthin by Chlorella zofingiensis Grown Under Limited Light Intensity

The fresh water green microalga Chlorella zofingiensisis known to accumulate ketocarotenoids – primarily astaxanthin but also canthaxanthin – when grown under stress conditions of high light irradiance and low nitrogen. We found that salt stress can replace light stress with respect to inducing carotenoid production: cells of C. zofingiensis grown under low light irradiance and subjected to salt and low nitrogen stress accumulated higher amounts of total secondary carotenoids than those growing under high light and low nitrogen stress. Furthermore, C. zofingiensis growing under conditions of salt stress and low light accumulated higher amounts of canthaxanthin than astaxanthin. It is suggested that for canthaxanthin accumulation under salt stress, light is not a limiting factor, but for astaxanthin accumulation high light irradiance is mandatory. These results may be applied in the future for the commercial production of canthaxanthin by C. zofingiensis in systems in which light availability is poor.     

Optimization of astaxanthin production by Phaffia rhodozyma through factorial design and response surface methodology.

Sequential methodology based on the application of three types of experimental designs was used to optimize the astaxanthin production of the mutant strain 25-2 of Phaffia rhodozyma in shake flask cultures. The first design employed was a factorial design 2(5), where the factors studied were: pH, temperature, percent of inoculum, carbon and nitrogen concentrations, each one at two levels. This design was performed in two medium types: rich YM medium and minimal medium, based on date juice (Yucca medium). With this first design the most important factors were determined (carbon concentration and temperature) that were used in the second experimental strategy: the method of steepest ascent was applied in order to rapidly approach the optimum. Finally, a second-order response surface design was applied using temperature and carbon concentration as factors. The optimal conditions stimulating the highest astaxanthin production were: 19.7 degrees C temperature; 11.25 g l(-1) carbon concentration; 6.0 pH; 5% inoculum and 0.5 g l(-1) nitrogen concentration. Under these conditions the astaxanthin production was 8100 microg l(-1), 92% higher than the production under the initial conditions.     

Effects of different fungal elicitors on growth, total carotenoids and astaxanthin formation by Xanthophyllomyces dendrorhous

Six fungal elicitors prepared from Rhodotorula rubra, Rhodotorula glutinis, Panus conchatus, Coriolus versicolor, Mucor mucedo, Mortieralla alpina M-23 were examined to determine their effects on the growth, total carotenoids and astaxanthin formation by Xanthophyllomyces dendrorhous. The results showed that different fungal elicitor could cause diversely stimulating effects. Among the fungal elicitors tested, the M. mucedo elicitor concentration of 30 mg l(-1) promoted the biomass and total carotenoids yield most remarkably, resulting in 69.81+/-6.00% and 78.87+/-4.15% higher than the control, respectively. At the concentration of 30 mg l(-1), R. glutinis elicitor stimulated the highest astaxanthin yield with a 90.60+/-5.98% increase compared to the control. The R. rubra elicitor concentration of 30 mg l(-1) resulted in the optimal total carotenoids and astaxanthin content to be 42.24+/-0.49% and 69.02+/-0.72% higher than the control, respectively. At the concentration of 30 mg l(-1), R. rubra elicitor gave the highest increase in the ratio of astaxanthin in total carotenoids by 18.85+/-0.11% of the control.     

Pb2+胁迫对绿球藻(Chlorococcum sp.)的影响研究

研究了不同Pb²⁺浓度(0.1、1、10、50、100、200、400 mg/L)处理对绿球藻(Chlorococcum sp.)生长、形态结构及生理特性的影响。与对照BG11培养的绿球藻比较,Pb²⁺浓度≤50 mg/L条件下培养的绿球藻细胞壁无明显增厚,色素变化不大;而暴露到Pb²⁺浓度>50 mg/L条件下培养,绿球藻细胞壁明显增厚,蛋白核消失。低浓度Pb²⁺(0.1~10 mg/L)对绿球藻生长基本没有影响;浓度在50 mg/L时,绿球藻仍能维持一定的生长速率;但当Pb²⁺浓度≥100 mg/L时,绿球藻的生长受到显著抑制。绿球藻的Chl a+Chl b以及Chl a含量均随培养基中Pb²⁺浓度的升高而逐渐减少。绿球藻净光合作用强度随培养基中Pb²⁺浓度的增大而逐渐降低,Pb²⁺浓度≥100 mg/L时净光合作用强度检测不到;当Pb²⁺浓度<50 mg/L时,绿球藻呼吸作用强度逐渐升高,之后呼吸作用强度逐渐降低。在实验的浓度下,绿球藻的丙二醛(MDA)含量、超氧化物歧化酶(SOD)和过氧化物酶(POD)活性都随培养基中Pb²⁺浓度的升高而逐渐增强;过氧化氢酶(CAT)则随Pb²⁺浓度的增大酶活性先升高后降低。当Pb²⁺浓度≤100 mg/L时,绿球藻对Pb²⁺的去除率都在95%以上;之后逐渐降低,浓度到400 mg/L时仍然达56.7%。结果显示,绿球藻是一种耐受Pb²⁺胁迫的藻类,对铅的去除率也高,可以应用于含铅污水的处理。     

Enhanced production of free trans-astaxanthin by oxidative stress in the cultures of the green microalga Chlorococcum sp

Free trans-astaxanthin accumulated in the alga Chlorococcum sp. was markedly enhanced from 3.664 mg g⁻¹ cell dry weight to 5.724 mg g⁻¹ cell dry weight when the culture was supplemented with hydrogen peroxide (0.1 mM) under mixotrophic conditions of growth. After saponification, a total of 7.086 mg astaxanthin per g cell dry weight was achieved. Similarly, in heterotrophic cultures, the total astaxanthin content was increased from 1.034 mg g⁻¹ cell dry weight without H₂O₂ to 1.782 mg g⁻¹ cell dry weight with 0.1mM H₂O₂. Results indicate that hydrogen peroxide effectively induces the formation of free trans-astaxanthin in Chlorococcum sp.     

Production of ketocarotenoids by microalgae

Among the highly valued ketocarotenoids employed for food coloration, astaxanthin is probably the most important. This carotenoid may be produced biotechnologically by a number of microorganisms, and the most promising seems to be the freshwater flagellate Haematococcus pluvialis (Chlorophyceae), which accumulate astaxanthin in their aplanospores. Many physiological aspects of the transition of the flagellate into aplanospores have been described. Mixotrophic cultivation and suitable irradiance may result in fairly good yields (up to 40 mg/l; 43 mg/g cell dry weight) within a reasonable time, under laboratory conditions. In order to compete with synthetic astaxanthin, suitable scaling-up is required. However, large-scale production in open ponds has proved unsatisfactory because of severe contamination problems. A selective medium might overcome this difficulty. Further research for the development of suitable strains is thus warranted. Received: 8 July 1998 / Received revision: 12 November 1998 / Accepted: 14 November 1998