• Title/Summary/Keyword: phosphate starvation

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Systemic Statistical Optimization of Astaxanthin Inducing Methods in Haematococcus pluvialis cells -Statistical Optimization of Astaxanthin Production in Haematococcus

  • Kim, Sun-Hyoung;Jeong, Sung Eun;Hong, Seong-Joo;Lee, Choul-Gyun
    • Journal of Marine Bioscience and Biotechnology
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    • v.6 no.1
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    • pp.31-40
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    • 2014
  • The production of astaxanthin in the microalga Haematococcus pluvialis has been investigated using a sequential methodology based on the application of two types of statistical designs. The employed preliminary experiment was a fractional factorial design $2^6$ in which the factors studied were: excessive irradiance and nitrate starvation, phosphate deficiency, acetate supplementation, salt stress, and elevated temperature. The experimental results indicate that the amount of astaxanthin accumulation in the cells can be enhanced by excessive irradiance and nitrate starvation whereas the other factors tested did not yield any enhancement. In the subsequent experiment, a central composite design was applied with four variables, light intensity, nitrate, phosphate, and acetate, at five levels each. The optimal conditions for the highest astaxanthin production were found to be $1040{\mu}E/(m^2{\cdot}s)$ light intensity, 0.04 g/L nitrate, 0.31 g/L phosphate, 0.05 g/L acetate concentration.

Phosphate and Carbon Source Regulation of Alkaline Phosphatase and Phospholipase in Vibrio vulnificus

  • Oh, Wan-Seok;Im, Young-Sun;Yeon, Kyu-Yong;Yoon, Young-Jun;Kim, Jung-Wan
    • Journal of Microbiology
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    • v.45 no.4
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    • pp.311-317
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    • 2007
  • In this study, the effects of phosphate concentration and carbon source on the patterns of alkaline phosphatase (APase) and phospholipase (PLase) expression in Vibrio vulnificus ATCC 29307 were assessed under various conditions. The activities of these enzymes were repressed by excess phosphate (4 mM) in the culture medium, but this repression was reversed upon the onset of phosphate starvation in low phosphate defined medium (LPDM) containing 0.2 mM of phosphate at approximately the end of the exponential growth phase. The expressions of the two enzymes were also influenced by different carbon sources, including glucose, fructose, maltose, glycerol, and sodium acetate at different levels. The APase activity was derepressed most profoundly in LPDM containing fructose as a sole carbon source. However, the repression/derepression of the enzyme by phosphate was not observed in media containing glycerol or sodium acetate. In LPDM-glycerol or sodium acetate, the growth rate was quite low. The highest levels of PLase activity were detected in LPDM-sodium acetate, followed by LPDM-fructose. PLase was not fully repressed by high phosphate concentrations when sodium acetate was utilized as the sole carbon source. These results showed that multiple regulatory systems, including the phosphate regulon, may perform a function in the expression of both or either APase and PLC, in the broader context of the survival of V. vulnificus.

Studies on the activities of ALPase, ACPase, ATPase and accumulation of volutin granules upon growth phase in saccharomyces uvarum (Saccharomyces uvarum의 배양시기에 따른 ALPase, ACPase, ATPase 활성도와 volutin과립 축적량)

  • 이기성;최영길
    • Korean Journal of Microbiology
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    • v.23 no.2
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    • pp.90-100
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    • 1985
  • The present study was designed to investigate cellular regulation of phosphate metabolism between catabolically repressed and derepressed states in yeast (Saccharomyces uvarum). The activities of various phospatases and the contents of phosphate compounds were detected according to the culture phase and various phosphate concentrations. As the results, Saccharomyces uvarum derepressed many phosphate metabolizing enzymes such as alkaline phosphatase, acid phosphatase and ATPase more than ten fold simultaneously during catabolic repression (phospgate and sugar starvation). At the same state, the amounts of orthophosphate, nucleotidic labile phosphate and acid soluble polypgosphate were increased, compared to basal levels of normally cultivated cells. $Mg^{++}-stimulated$ type among all phospatases was appeared to have most of the enzyme activity. It could be postulated that $K^+ -stimulated$ alkaline phosphatase was directly or indirectly correlated with the synthesis of acid insoluble polyphosphate $Mg^{++}-stimulated$ phosphatase with the degradation of polyphosphates. In case of cultivation in the medium supplemented with sugar and phosphate (catabolic derepression), phospgatase activities except for alkaline phosphatase were decreased rapidly through the progressive batch culture, After 12 hrs culture, at early exponential phase, the cellular accumulation of acid insoluble polyphosphate increased about 5 fold, compared to those of the starved cells. Under catabolic repression, it could be postulated that intracellular phosphate metabolism was regulated by derepressions of phosphatases. The function of polyphosphate system was shown to compensate the ATP/ADP system as phosphate donor and energy source especially during catabolic repression.

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Mineral- and Tissue-Specific Metabolic Changes in Tomato (Lycopersicon esculentum L.) Plants Grown under NPK-Starved Conditions

  • Sung, Jwakyung;Lee, Yejin;Lee, Seulbi;Lim, Jungeun;Lee, Deogbae
    • Korean Journal of Soil Science and Fertilizer
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    • v.49 no.6
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    • pp.689-698
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    • 2016
  • Specific metabolic network responses to mineral starvation are not well-defined. We examined a detailed broad-scale identification of metabolic responses of tomato leaf and root to N, P or K starvation. Tomato plants were grown hydroponically under optimal (5 mM N, 0.5 mM P, or 5 mM K) and starved (0.5 mM N, 0.05 mM P, or 0.5 mM K) conditions and metabolites were measured by LC-MS and GC-MS. Overall, the levels of metabolites (lipids, nucleotides, peptides and secondary metabolites) presented in this paper largely showed mineral- and tissue-specific responses. Most strikingly, G3P (glycerol-3-P), GPC (glycerol-P-choline) and choline phosphate responded differently to a type of mineral; an increase in N or K starvation and a decrease in P starvation. A dramatic increase in the levels of secondary metabolites, in particular, rutin and chlorogenate in both tomato tissues during N starvation were observed. Based on these data, it is necessary to clearly elucidate an unknown event taking place in a variety of abiotic impacts, and we are now studying to expand our knowledge on metabolic- and proteomic-responses using GS-MS and LC-MS.

Effect of Nutrient Limitation on Lipid Content and Fatty Acid Composition of Mutant Chlamydomonas reinhardtii (돌연변이 Chlamydomonas reinhardtii의 영양분 제한에 따른 지질 생산 및 지방산 조성 변화 연구)

  • Baek, Jaewon;Choi, Jong-il
    • KSBB Journal
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    • v.30 no.2
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    • pp.91-95
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    • 2015
  • Production of biodiesel from microalgae is dependent on the microalgal lipid content and free fatty acid composition. Both lipid and free fatty acid are regulated by nutrient sources. In this study, newly developed mutant Chlamydomonas reinhardtii with higher lipid content was investigated for the effect of nutrient limitation. Nitrogen $NO_3{^{-}}$ and phosphate $PO_4{^{3-}}$ were limited for nutrient starvation during the cultivation. Under nutrient starvation, total lipid content level was increased to 27~33% and C16:0 fatty acid content constituted over 31~43% of total fatty acid. Interestingly, we also found that the expression of fatty acid desaturase (FAD7) was decreased when nutrients were starved.

A Study on The Content of Liver Protein, Nucleic Acids, and Guanine Deaminase Activity of Mouse During Acute Starvation (급성(急性) 기아(饑餓)마우스의 간단백질(肝蛋白質), 핵산(核酸) 및 Guanine Deaminase 활성(活性)에 관(關)한 연구(硏究))

  • Park, Seung-Hee;Kim, Seung-Won
    • Journal of Nutrition and Health
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    • v.1 no.2
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    • pp.107-115
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    • 1968
  • Number of aspects, not only nutritional but social as well as political involved in human starvation pose nowadays global problems. In order to help establish the minimum nutritional requirements in the daily life of a man and to free people as well from either undernourishment, malnutrition or even starvation many workers have devoted themselves so far on the research programs to know what and how number of metabolic events take place in animals in vivo. It is the purpose of the present paper to examine in effect to what extent both of the protein and nucleic acids (DNA & RNA) together with an enzyme, guanine deaminase, which converts guanine into xanthine and in turn ends up to uric acid as an end product, undergo changes, quantitatively during acute starvation, using the mouse as an experimental animal. The mouse was strictly inhibited from taking foods except drinking water ad libitum and was sacriflced 24, 48, and 72 hours following starvation thus acutely induced. The animals consisted of two experimental groups, one control and another starvation groups, each being consisted of 6-24 mice of whose body weights ranged in the vicinity of 10 g. The animals were sacriflced by a blow on the head, followed by immediate excision of their livers into ice-cold distilled water, washing adherent blood and other contaminant tissues. The liver was minced foramin, by an all-glass homogenizer immersing it in an ice-bath, followed by subsequent fractionatin of the homogenate (10% W/V in 0.25M sucrose solution made up with 0.05M phosphate buffer of pH 7.4). For the liver protein and guanine deaminase assay, the 10% homogenate was centrifuged at 600 x g for 10 minutes to eliminate the nuclear fraction; and for the estimation of DNA and RNA, the homogenate was prepared by the addition of 10% trichloroacetic acid in order to free the homogenate from the acid-soluble fraction, the remaining residue being delipidate by the addition of alcohol and dried in vacuo for later KOH (IN) hydrolysis. The changes in body and liver wegihts during acute starvation were checked gravimetrically. Protein contents in the liver were monitored by the method of Lowry et al; and guanine deaminase activities were followed by the assay of liberated ammonia from the substrate utilizing the Caraway's colorimetry. The extraction of both DNA and RNA was performed by the Schmidt-Thannhauser's method, which was followed by Marmur's method of purification for DNA and by Chargaff's method of purification for RNA. The determinations of both DNA and RNA were carried out by the diphenylamine reaction for the former and by the orcinol reaction for the latter. The following resume was the results of the present work. 1. It was observed that the body as well as liver weights fall abruptly during starvation, and that the loss of body weight showed no statistical correlation with the decreases in the content of liver protein. 2. The content of liver protein and activity of liver guanine deaminase activity as well decline dramatically, and the specific activities of the enzyme (activity/protein), however, decreased gradually as starvation proceeded. 3. Both of the nucleic acids, DNA and RNA, showed decrements in the liver of mouse during acute starvation; the latter, however, being more striking in the decline as compared to the former. 4. The decreases in the liver protein content as resulted from the acute starvation had no statistically significant correlation with the decrements of DNA in the same tissue, but had regressed with a significant statistical correlation with the fall of RNA in the tissue. 5. The decrease in the activity of guanine deaminase in the liver of mouse during acute starvation was functionally more proportional to the decrease in RNA than DNA, and moreover correlated with the changes in the content of the liver protein. 6. The possible mechanisms involved during in this acute starvation as bring the decreases in the contents of DNA, protein, and guanine deaminase were discussed briefly.

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Differential Stringent Responses of Streptomyces coelicolor M600 to Starvation of Specific Nutrients

  • Ryu, Yong-Gu;Kim, Eun-Sook;Kim, Dae-Wi;Kim, Sung-Keun;Lee, Kye-Joon
    • Journal of Microbiology and Biotechnology
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    • v.17 no.2
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    • pp.305-312
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    • 2007
  • This study focused on the involvement of the unusual nucleotide (p)ppGpp, a stringent factor, during the morphological and physiological differentiation of Streptomyces coelicolor. Two genes, relA and rshA, were disrupted to demonstrate the roles of the stringent factor in the differentiation. The intracellular concentration of (p)ppGpp in the wild-type (M600) and disrupted mutants was measured in relation to the intentional starvation of a specific nutrient, such as carbon, nitrogen, and phosphate or the in situ depletion of nutrients in a batch culture. As a result, it was found that the morphological characteristic of the ${\Delta}relA$ mutant was a bld phenotype forming condensed mycelia, whereas the ${\Delta}rshA$ mutant grew fast-forming spores and straightforward mycelia. In both mutants, the production of actinorhodin (Act) was completely abolished, yet the undecylprodigiosin (Red) production was increased. Intracellular (p)ppGpp was detected in the ${\Delta}relA$ mutant in the case of limited phosphate, yet not with limited carbon or nitrogen sources. In contrast, (p)ppGpp was produced in the ${\Delta}rshA$ mutant under limited carbon and nitrogen conditions. Therefore, (p)ppGpp in S. coelicolor was found to be selectively regulated by either the RelA or RshA protein, which was differentially expressed in response to the specific nutrient limitation. These results were also supported by the in situ ppGpp production during a batch culture. Furthermore, it is suggested that RelA and RshA are bifunctional proteins that possess the ability to both synthesize and hydrolyze (p)ppGpp.

Biochemical and Molecular Analysis of OsPAP1: A Phosphate Starvation Induced Purple Acid Phosphatase Gene from Rice

  • Hur, Yeon Jae;Yi, Young Byung;Kim, Tae Ho;Kim, Doh Hoon
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.455-462
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    • 2010
  • Purple acid phosphatase is important for phosphorus remobilization in plants, but its role in plant adaptation to low phosphorus availability is not known. The cDNA encoding O. sativa purple acid phosphatase (OsPAP1) has 1008 bp with an open reading frame of 335 amino acid residues. The amino acid sequence of OsPAP1 cDNA shows of 50-51% identity with other plant purple acid phosphatases. OsPAP1 was expressed in rice plants and in cell cultures in the absence of phosphate ($P_i$). The expression was organ-specific with the strongest expression in $P_i$-deprived roots. Functional expression of the OsPAP1 gene in the transgenic Arabidopsis line was confirmed by northern and western blot analysis. OsPAP1 overexpression lines had higher phosphatase activity than wild-type. Overexpression of OsPAP1 in Arabidopsis plants resulted in increased Pi accumulation under Pi sufficient condition. These results show that the OsPAP1 gene represents more efficient $P_i$ uptake and can be used to develop new transgenic dicotyledonous plants.