Isolation and characterization of a stress-inducible Dunaliella salina Lcy-β gene encoding a functional lycopene β-cyclase

Ramos, A.; Coesel, Sacha; Marques, Ana Rita; Rodrigues, Marta V.; Baumgartner, Alexandra; Noronha, João Paulo; Rauter, Amélia P.; Brenig, Bertram; Varela, João

doi:10.1007/s00253-008-1492-4

Cited by 70 publications

(53 citation statements)

References 44 publications

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“…-carotene content mainly followed the same tendency [COESEL et al, 2008]. Our data on -carotene accumulation in the free of nutrients culture media confirmed that pattern.This molecular data brought the authors to the conclusion that nutrient depletion is necessary and sufficient for induction of -carotene accumulation in D. salina cells at transcriptional level [COESEL et al, 2008]. Our data add to this finding that nutrient…”

supporting

confidence: 86%

“…This molecular data brought the authors to the conclusion that nutrient depletion is necessary and sufficient for induction of -carotene accumulation in D. salina cells at transcriptional level [COESEL et al, 2008]. Our data add to this finding that nutrient…”

Section: Discussionsupporting

confidence: 63%

“…Some researchers attach much importance to the impact of salinity and irradiance [LOEBLICH, 1982; BOROWITZKA, 2013], some other -to nutrient depletion [LERCHE, 1936[LERCHE, /1937COESEL et al, 2008]. Not all experiments confirm the effect of phosphorus deficiency [BEN-AMOTZ et al, 1982].…”

Section: Evaluation Of Contribution Of Salinity Irradiance and Nutrmentioning

confidence: 99%

“…Cited studies [COESEL et al, 2008;RAMOS et al, 2008;2009] showed that factorsconditions interacted with factors-resources at transcriptional level. Signal transduction network might be supposed, providing interactions of the signals and modulation of the effect.…”

Section: Evaluation Of Contribution Of Salinity Irradiance and Nutrmentioning

confidence: 99%

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Evaluation of contribution of salinity, irradiance, and nutrient deficiency into the yield of cells and b-carotene accumulation in the culture of Dunaliella salina (Chlorophyta)

Victoria¹,

Katerina²,

Oleksandr³

2018

Chornomors’k. bot. z.

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Evaluation of contribution of salinity, irradiance, and nutrient deficiency into the yield of cells and -caroteneThe purpose of the study was to evaluate the contribution of salinity, irradiance, nitrate and phosphate, and their interactions into the yield of cell number and -carotene accumulation in Dunaliella salina. To avoid confounding of the effects of factors-conditions by the depletion of factors-resources, the alga was grown in fed-batch culture. In the level ranges of the experimental factors (irradiance 2-8 klx, salinity 1-4 M NaCl, KNO3 0-80 mg L-1, K2HPO4 0-10 mg L-1), nitrate and phosphate influenced the productivity of culture by cell number and -carotene accumulation more strongly than salinity and irradiance. Effects of salinity and irradiance depended on nutrients and their pre-supply to the inoculum. Total effect size  2 of nutrients on cell yield comprised 0,59 for non-starved and 0,43 for starved inoculum, whereas total effect size of factors-conditions -0,10 and 0,12 correspondingly. As to cellular -carotene content, total effect size of nutrients on the cells grown from nonstarved and starved inoculum was 0,71 and 0,58, and of factors conditions -0,8 and 0,5 correspondingly. Remained variances of cell yield and -carotene content were attributed to the interactions of salinity and irradiance with the nutrients. The combination of high values of salinity and irradiance exerted its own, unconfounded by depletion of nutrients, but lower influence on -carotene accumulation. The highest -carotene content of 53 pg per cell was observed in the culture grown from the starved inoculum at the deficiency of phosphorus. Combination of high salinity and irradiance values yielded 17 pg of -carotene per cell compared to about 5 pg under the optimal culture conditions. Controll nutrient supply would be the most powerful tool for biosynthesis control in D. salina culture.Key words: nitrate, phosphate, microalga, cultivation methodology КОМАРИСТА В.П., БІЛОУСОВА К.М., РУДАСЬ О.М. (2018). Кількісне визначення внеску солоності, освітленості та дефіциту біогенів у вихід клітин і накопичення -каротину в культурі Dunaliella salina (Chlorophyta). Чорноморськ. бот. ж., 14 (1): 43-55. doi: 10.14255/2308-9628/18.141/4 Метою дослідження було кількісне визначення внеску солоності, освітленості, нітрату, фосфату та їхної взаємодії до врожаю Dunaliella salina за кількістю клітин та накопиченням -каротину. Щоб запобігти змішенню ефектів факторів-умов з вичерпанням факторів-ресурсів, водорість вирощували в періодичній культурі з підживленням. В дослідженому діапазоні факторів (освітленість 2-8 клк, солоність 1-4 M NaCl, KNO3 0-80 мг/л, K2HPO4 0-10 мг/л), нітрат і фосфат сильніше впливали на продуктивність культури за кількістю клітин та накопиченням -каротину, ніж солоність та освітленість. Вплив солоності та освітленості залежав від біогенів та  © Komaristaya V.P., Bilousova K.M., Rudas O.M. Чорноморськ. бот. ж., 14 (1): 43-55.Чорноморський ботанічний журнал -том 14, № 1 (2018) Komaristaya V.P., Bilousova...

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supporting

confidence: 86%

Section: Discussionsupporting

confidence: 63%

Section: Evaluation Of Contribution Of Salinity Irradiance and Nutrmentioning

confidence: 99%

Section: Evaluation Of Contribution Of Salinity Irradiance and Nutrmentioning

confidence: 99%

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Evaluation of contribution of salinity, irradiance, and nutrient deficiency into the yield of cells and b-carotene accumulation in the culture of Dunaliella salina (Chlorophyta)

Victoria¹,

Katerina²,

Oleksandr³

2018

Chornomors’k. bot. z.

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“…NRC-1 (19), Haloarcula marismortui (18), Halobacterium salinarum (22), uncultured marine bacterium 66A03 (16), and uncultured marine bacterium HF10 49E08 (21). ␤-Carotene biosynthetic genes crtE, crtB, crtI, crtY, ispA, and idi encode the enzymes necessary for the synthesis of ␤-carotene from isopentenyl diphosphate, and the Idi, IspA, CrtE, CrtB, CrtI, and CrtY proteins have been characterized in vitro (23)(24)(25)(26)(27)(28). Blh protein has been proposed to catalyze or regulate the conversion of ␤-carotene to retinal (29, 30), but there is no direct proof of the enzymatic activity.…”

mentioning

confidence: 99%

In Vitro Characterization of a Recombinant Blh Protein from an Uncultured Marine Bacterium as a β-Carotene 15,15′-Dioxygenase

Kim

Yeom

et al. 2009

Journal of Biological Chemistry

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Codon optimization was used to synthesize the blh gene from the uncultured marine bacterium 66A03 for expression in Escherichia coli. The expressed enzyme cleaved ␤-carotene at its central double bond (15,15) to yield two molecules of all-transretinal. The molecular mass of the native purified enzyme was ϳ64 kDa as a dimer of 32-kDa subunits. The K m , k cat , and k cat /K m values for ␤-carotene as substrate were 37 M, 3.6 min ؊1 , and 97 mM ؊1 min ؊1 , respectively. The enzyme exhibited the highest activity for ␤-carotene, followed by ␤-cryptoxanthin, ␤-apo-4-carotenal, ␣-carotene, and ␥-carotene in decreasing order, but not for ␤-apo-8-carotenal, ␤-apo-12-carotenal, lutein, zeaxanthin, or lycopene, suggesting that the presence of one unsubstituted ␤-ionone ring in a substrate with a molecular weight greater than C 35 seems to be essential for enzyme activity. The oxygen atom of retinal originated not from water but from molecular oxygen, suggesting that the enzyme was a ␤-carotene 15,15-dioxygenase. Although the Blh protein and ␤-carotene 15,15-monooxygenases catalyzed the same biochemical reaction, the Blh protein was unrelated to the mammalian ␤-carotene 15,15-monooxygenases as assessed by their different properties, including DNA and amino acid sequences, molecular weight, form of association, reaction mechanism, kinetic properties, and substrate specificity. This is the first report of in vitro characterization of a bacterial ␤-carotene-cleaving enzyme.Vitamin A (retinol) is a fat-soluble vitamin and important for human health. In vivo, the cleavage of ␤-carotene to retinal is an important step of vitamin A synthesis. The cleavage can proceed via two different biochemical pathways (1, 2). The major pathway is a central cleavage catalyzed by mammalian ␤-carotene 15,15Ј-monooxygenases (EC 1.14.99.36). ␤-Carotene is cleaved by the enzyme symmetrically into two molecules of alltrans-retinal, and retinal is then converted to vitamin A in vivo (3-5). The second pathway is an eccentric cleavage that occurs at double bonds other than the central 15,15Ј-double bond of ␤-carotene to produce ␤-apo-carotenals with different chain lengths, which are catalyzed by carotenoid oxygenases from mammals, plants, and cyanobacteria (6). These ␤-apo-carotenals are degraded to one molecule of retinal, which is subsequently converted to vitamin A in vivo (2).␤-Carotene 15,15Ј-monooxygenase was first isolated as a cytosolic enzyme by identifying the product of ␤-carotene cleavage as retinal (7). The characterization of the enzyme and the reaction pathway from ␤-carotene to retinal were also investigated (4, 8). The enzyme activity has been found in mammalian intestinal mucosa, jejunum enterocytes, liver, lung, kidney, and brain (5, 9, 10). Molecular cloning, expression, and characterization of ␤-carotene 15,15Ј-monooxygenase have been reported from various species, including chickens (11), fruit flies (12), humans (13), mice (14), and zebra fishes (15).Other proteins thought to convert ␤-carotene to retinal include bacterioopsin-r...

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Carotenoid and fatty acid metabolism in light‐stressed Dunaliella salina

Lamers

Laak

Kaasenbrood

et al. 2010

Biotech & Bioengineering

239

127

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beta-Carotene is overproduced in the alga Dunaliella salina in response to high light intensities. We have studied the effects of a sudden light increase on carotenoid and fatty acid metabolism using a flat panel photobioreactor that was run in turbidostat mode to ensure a constant light regime throughout the experiments. Upon the shift to an increased light intensity, beta-carotene production commenced immediately. The first 4 h after induction were marked by constant intracellular levels of beta-carotene (2.2 g LCV(-1)), which resulted from identical increases in the production rates of cell volume and beta-carotene. Following this initial phase, beta-carotene productivity continued to increase while the cell volume productivity dropped. As a result, the intracellular beta-carotene concentration increased reaching a maximum of 17 g LCV(-1) after 2 days of light stress. Approximately 1 day before that, the maximum beta-carotene productivity of 30 pg cell(-1) day(-1) (equivalent to 37 mg LRV(-1) day(-1)) was obtained, which was about one order of magnitude larger than the average productivity reported for a commercial beta-carotene production facility, indicating a vast potential for improvement. Furthermore, by studying the light-induced changes in both beta-carotene and fatty acid metabolism, it appeared that carotenoid overproduction was associated with oil globule formation and a decrease in the degree of fatty acid unsaturation. Our results indicate that cellular beta-carotene accumulation in D. salina correlates with accumulation of specific fatty acid species (C16:0 and C18:1) rather than with total fatty acid content.

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Isolation and characterization of a stress-inducible Dunaliella salina Lcy-β gene encoding a functional lycopene β-cyclase

Cited by 70 publications

References 44 publications

Evaluation of contribution of salinity, irradiance, and nutrient deficiency into the yield of cells and b-carotene accumulation in the culture of Dunaliella salina (Chlorophyta)

Evaluation of contribution of salinity, irradiance, and nutrient deficiency into the yield of cells and b-carotene accumulation in the culture of Dunaliella salina (Chlorophyta)

In Vitro Characterization of a Recombinant Blh Protein from an Uncultured Marine Bacterium as a β-Carotene 15,15′-Dioxygenase

Carotenoid and fatty acid metabolism in light‐stressed Dunaliella salina

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