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环境条件对淡水硅藻美丽星杆藻(硅藻纲)培养物中各种酶活性和三酰甘油含量的影响

Effect of environmental conditions on various enzyme activities and triacylglycerol contents in cultures of the freshwater diatom, Asterionella formosa (Bacillariophyceae).

作者信息

Mekhalfi Malika, Amara Sawsan, Robert Sylvie, Carrière Frédéric, Gontero Brigitte

机构信息

Aix Marseille Université, CNRS, UMR 7281 Laboratoire de Bioénergétique et Ingénierie des Protéines, 31 Chemin Joseph Aiguier, 13402 Marseille Cedex 20 France.

UMR 7282 Enzymology at Interfaces and Physiology of Lipolysis, 31 Chemin Joseph Aiguier, 13402 Marseille Cedex 20 France.

出版信息

Biochimie. 2014 Jun;101:21-30. doi: 10.1016/j.biochi.2013.12.004. Epub 2013 Dec 17.

Abstract

A detailed analysis of triacylglycerols (TAGs) contents, fatty acid patterns and key enzyme activities in the freshwater diatom Asterionella formosa was performed under various conditions, including nitrate, iron and silicon limitation (stress conditions), or bicarbonate and phytohormones supplementation (stimulation conditions). Of all the conditions tested, the addition of bicarbonate produced the greatest increase (5-fold) in TAGs contents compared to the control while the biomass increased. The addition of phytohormones also allowed a significant increase in TAGs of about 3-fold while the biomass increased. Silicon, unlike iron and nitrate limitation, also triggered a significant increase in TAGs contents of 3.5-fold but negatively affected the biomass. Analysis of fatty acid profiles showed that the mono-unsaturated C16:1 fatty acid was the most abundant in A. formosa, followed by C16:0, C14:0 and eicosapentaenoic acid (EPA; C20:5 n-3). EPA levels were found to increase under nitrate and iron limitation. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), phosphoribulokinase (PRK), phosphofructokinase (PFK), glucose-6-phosphate dehydrogenase (G6PDH) and malate dehydrogenase (MDH) activities differed with growth conditions. Most enzymes were up-regulated in stimulated cells while in the case of stressed cells, the pattern of activities was more variable. Detailed analysis of all enzyme activities showed that the most important enzyme among those tested was GAPDH which could be a good candidate for genetic engineering of high lipid-producing algae. This study provides a better understanding of key enzymes and biochemical pathways involved in lipid accumulation processes in diatoms.

摘要

在各种条件下,包括硝酸盐、铁和硅限制(胁迫条件),或补充碳酸氢盐和植物激素(刺激条件),对淡水硅藻美丽星杆藻中的三酰甘油(TAGs)含量、脂肪酸模式和关键酶活性进行了详细分析。在所有测试条件中,与对照相比,添加碳酸氢盐使TAGs含量增加最多(5倍),同时生物量增加。添加植物激素也使TAGs显著增加约3倍,同时生物量增加。与铁和硝酸盐限制不同,硅也引发了TAGs含量3.5倍的显著增加,但对生物量有负面影响。脂肪酸谱分析表明,单不饱和C16:1脂肪酸在美丽星杆藻中含量最高,其次是C16:0、C14:0和二十碳五烯酸(EPA;C20:5 n-3)。发现EPA水平在硝酸盐和铁限制下会增加。甘油醛-3-磷酸脱氢酶(GAPDH)、磷酸核酮糖激酶(PRK)、磷酸果糖激酶(PFK)、葡萄糖-6-磷酸脱氢酶(G6PDH)和苹果酸脱氢酶(MDH)的活性随生长条件而不同。大多数酶在受刺激的细胞中上调,而在胁迫细胞中,活性模式变化更大。对所有酶活性的详细分析表明,所测试的酶中最重要的是GAPDH,它可能是高产脂藻类基因工程的良好候选者。这项研究有助于更好地理解硅藻脂质积累过程中涉及的关键酶和生化途径。

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