本文選題：原始小球藻　切入點：生物量　出處：《湘潭大學》2016年碩士論文　論文類型：學位論文

【摘要】：本研究考察了不同氮磷耦合誘導模式(即富氮-富磷、貧氮-富磷、富氮-貧磷、貧氮-貧磷、貧氮-磷限制)對小球藻(Chlorella protothecoides)生物量及油脂積累的影響,并考察了最佳氮磷耦合誘導模式下小球藻細胞油脂高效富集的調(diào)控機制。結(jié)果表明,富氮-富磷耦合誘導模式下小球藻最大生物量為5.92 g/L,高于貧氮-富磷(2.99 g/L)、富氮-貧磷(2.38 g/L)、貧氮-貧磷(0.76 g/L)及貧氮-磷限制(2.81 g/L)耦合誘導模式的生物量;貧氮-貧磷模式下,小球藻最大油脂含量為55.8%,優(yōu)于富氮-富磷(22.17%)、貧氮-富磷(52.5%)、富氮-貧磷(32.77%)及貧氮-磷限制(53.7%)耦合誘導模式的油脂含量;貧氮-富磷模式下,最大油脂產(chǎn)率為224.14 mg/L/d,這一油脂產(chǎn)率分別是對照組、富氮-貧磷、貧氮-貧磷、貧氮-磷限制耦合誘導模式的1.19~3.70倍。這證實貧氮-富磷耦合模式是促進小球藻(C.Protothecoides)細胞油脂積累的潛力誘導模式。進一步地,采用二維熒光差異凝膠電泳定量蛋白質(zhì)組學手段揭示貧氮-富磷耦合誘導模式下小球藻細胞油脂積累相關(guān)調(diào)控機制。采用貧氮-富磷耦合模式誘導的藻細胞中,鑒定出27個差異表達蛋白質(zhì),包括8個下調(diào)蛋白和19個上調(diào)蛋白。根據(jù)代謝通路分析得出,糖酵解途徑中尿苷二磷酸-葡萄糖-6-脫氫酶和葡萄糖-1-磷酸腺苷轉(zhuǎn)移酶是貧氮-富磷耦合誘導模式下小球藻細胞油脂富集的關(guān)鍵調(diào)控因子,這兩種酶可催化細胞中的淀粉轉(zhuǎn)化為葡萄糖,為糖酵解途徑的開始提供切入點;磷酸戊糖途徑(PPP)中表達上調(diào)的轉(zhuǎn)酮醇酶將五碳酮糖和醛糖轉(zhuǎn)化為七碳酮糖和3-磷酸甘油醛(G3P);二磷酸核酮糖鹽羧化酶(RuBis CO)是提高卡爾文循環(huán)的關(guān)鍵酶,這種酶表達上調(diào)可以促進3-磷酸甘油醛(G3P)進入糖酵解途徑;蘋果酸脫氫酶(ME)表達上調(diào)能有效提高小球藻細胞油脂產(chǎn)率,可據(jù)此推測關(guān)鍵節(jié)點調(diào)控的脂質(zhì)合成積累路徑;氮饑餓信號調(diào)控因子—硝酸還原酶(NR)是小球藻細胞油脂產(chǎn)率提高的潛在調(diào)控因子;谷氨酸鹽脫氫酶(GLDH)表達上調(diào)可有效抵御培養(yǎng)條件失衡。這些研究發(fā)現(xiàn)為人工干預微藻油脂積累、推進微藻生物柴油規(guī)�；l(fā)展提供基礎,同時也為其它藻類油脂代謝研究提供參考。綜上所述,貧氮-富磷耦合模式是促進小球藻細胞油脂積累的有效誘導方式,這一模式為實現(xiàn)微藻高油脂產(chǎn)量提供了可能。而這些調(diào)控小球藻油脂積累的關(guān)鍵酶的發(fā)現(xiàn)為建立高效產(chǎn)油機制和基于微藻制備生物柴油的工業(yè)化生產(chǎn)奠定了基礎。
[Abstract]:In this study, the effects of different nitrogen and phosphorus coupling induction models (i.e., rich in nitrogen and phosphorus, low in nitrogen and phosphorus, rich in nitrogen and phosphorus, low in nitrogen and phosphorus, and low in nitrogen and phosphorus) on the biomass and oil accumulation of Chlorella protothecoides were investigated. The regulation mechanism of oil enrichment in Chlorella vulgaris cells under the optimal nitrogen and phosphorus coupling induction model was investigated. The maximum biomass of Chlorella vulgaris was 5.92 g / L, which was higher than 2.99 g / L of nitrogen and phosphorus rich, 2.38 g / L, 0.76 g / L and 2.81 g / L of N / P and 2.81 g / L, respectively. The maximum oil content of Chlorella vulgaris was 55.8, which was better than that of nitrogen-rich phosphate-rich chlorella 22.17, nitrogen-rich phosphorus-rich 52.5, nitrogen-poor phosphorus-rich (32.77) and nitrogen-phosphorus limiting (53.775)) -induced oil content, while in the nitrogen-phosphorus-rich model, The maximum oil yield is 224.14 mg / L / d, which is the control group, which is rich in nitrogen and poor in phosphorus, low in nitrogen and poor in phosphorus. The results show that the coupling model is a potential model for promoting the accumulation of lipid in C. Protothecoides cells. The regulation mechanism of lipid accumulation in Chlorella vulgaris cells induced by N-rich coupling was revealed by two-dimensional fluorescence differential gel electrophoresis (FDGE) quantitative proteomics, and the effect of nitrogen-phosphorus-rich coupling model on lipid accumulation in Chlorella vulgaris cells was investigated. Twenty-seven differentially expressed proteins were identified, including 8 down-regulated proteins and 19 up-regulated proteins. In glycolysis pathway, uridine diphosphate glucose-6-dehydrogenase and glucose-1-phosphate transferase are the key regulating factors of lipid enrichment in Chlorella vulgaris cells under the coupling induction of nitrogen and phosphorus deficiency. These two enzymes can catalyze the conversion of starch in cells to glucose, which provides a starting point for the beginning of glycolysis pathway. The up-regulated transketol enzyme expressed in the pentose phosphate pathway (PPP) converts pentacarbonose and aldose to heptose and glyceraldehyde 3-phosphate (G3PN), and ribonose diphosphate carboxylase (RuBis COS) is the key enzyme to improve the Calvin cycle. The up-regulation of the enzyme expression can promote the glycolysis pathway of glyceraldehyde-3-phosphate (G3P3), and the increase of malate dehydrogenase (MEE) expression can effectively increase the oil production rate of Chlorella vulgaris cells, which can be used to speculate the accumulation pathway of lipid synthesis regulated by the key nodes. Nitrogen starvation signal regulator, nitrate reductase (NR), is a potential regulator for the increase of oil production in Chlorella vulgaris cells. The up-regulated expression of glutamate dehydrogenase (GLDH) can effectively resist the imbalance of culture conditions. These findings provide a basis for the artificial intervention of microalgae oil accumulation and the promotion of large-scale development of microalgae biodiesel. In conclusion, the coupling model of nitrogen-rich phosphorus is an effective inductive way to promote the accumulation of lipid in Chlorella vulgaris cells. The discovery of these key enzymes to regulate the accumulation of Chlorella microalgae oil may provide a basis for the establishment of efficient oil production mechanism and the industrialized production of biodiesel based on microalgae.
【學位授予單位】：湘潭大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：TE667

【相似文獻】

相關(guān)期刊論文 前10條

1 ;富磷集團制定“九五”及2010年發(fā)展目標[J];硫酸工業(yè);1996年03期

2 李寶華,陳尚,朱明遠;圍隔生態(tài)系富磷及油污染對葉綠素a變化的影響[J];黃渤海海洋;2001年04期

3 宋新杰,胡青燕,田恒水;洗滌劑富磷污染及其對策[J];化工環(huán)保;1998年01期

4 左寧;吉芳英;;外排富磷污水的化學固磷與磷回收潛能[J];中國給水排水;2011年19期

5 李佳鳳;呂錫武;徐微;蔣彬;;誘導結(jié)晶反應器回收富磷上清液中磷的研究[J];中國給水排水;2010年01期

6 周寒梅;包燕平;林路;;P_2O_5對CaO-SiO_2-Fe_tO-P_2O_5渣中富磷相的影響[J];中國冶金;2013年01期

7 陸敏博;王世和;;富磷上清液鐵接觸除磷效果預測模型[J];東南大學學報(自然科學版);2009年02期

8 鄒鴻祥;;電爐法生產(chǎn)黃磷的廢渣污染現(xiàn)狀及治理[J];無機鹽工業(yè);1985年06期

9 陸敏博;王世和;;富磷上清液鐵接觸除磷污泥性質(zhì)與資源化利用[J];環(huán)境工程學報;2009年09期

10 陸敏博;王世和;;富磷上清液鐵接觸除磷工藝及影響因素[J];環(huán)境工程學報;2008年09期

相關(guān)重要報紙文章 前4條

1 崔紅秀;富磷礦突出“重圍”[N];中國礦業(yè)報;2002年

2 ;宜昌富磷公司效益創(chuàng)新高[N];中國礦業(yè)報;2003年

3 崔紅秀;富磷礦業(yè)著力培養(yǎng)青年隊伍[N];中國礦業(yè)報;2003年

4 王一凡;磷還夠我們用多少年[N];中國石油報;2004年

相關(guān)博士學位論文 前4條

1 閻凱;滇中富磷區(qū)典型植物群落氮磷化學計量格局與徑流輸出特征[D];云南大學;2015年

2 何鋒;滇池流域富磷區(qū)磷流失特征及控磷植物群落恢復研究[D];云南大學;2015年

3 王新宇;富磷水體中鈾的賦存形態(tài)與分配研究[D];成都理工大學;2014年

4 吉方英;排除厭氧富磷污水ERP-SBR除磷脫氮工藝研究[D];重慶大學;2004年

相關(guān)碩士學位論文 前6條

1 韓方欣;貧氮-富磷耦合誘導小球藻油脂積累及調(diào)控機制[D];湘潭大學;2016年

2 于宗赫;基于投加海水的化學沉淀法在富磷上清液除磷中的應用[D];中國海洋大學;2006年

3 楊琴;富磷污泥釋磷能力及磷的回收潛能研究[D];重慶大學;2002年

4 施政廷;滇池流域山地富磷區(qū)不同植被條件下面源污染及其輸出特征研究[D];云南大學;2015年

5 陳愛華;富磷條件對磷化銦晶體缺陷及Fe激活率的影響[D];河北工業(yè)大學;2014年

6 高海鳳;富磷條件下生長的InP單晶體材料特性研究[D];河北工業(yè)大學;2011年

，

本文編號：1591914