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標題: 建構無CO2之光生化產H2綠色工場-於綠藻去氧突變株過度表現產氫
Generation of a Zero-CO/sub 2/-Emission Green Factory for Photo-Biological H/sub 2/ Production-Overexpression of Hydrogenase in O/sub 2/-Deprived Green Algal Chlorella
作者: 簡麗鳳
關鍵字: 商品化;Chlorella;能源工程, 生物科學類;小球藻;氫氣;RNA 干擾;光系統II;電子傳遞鏈;放氧複合體;產氫?;hydrogen;RNA interference;photosystem II;electron transport;D1protein;oxygen evolving complex;overexpression;hydrogenase;small-scale bioreactor
A zero-CO2-emission fuel is urgently required to meet the energy demand and to control theeffects of global warming. Solar energy is the most abundant renewable energy source andcapable of powering large scale clean fuel production. Hydrogen (H2) is considered to be oneof the most promising clean fuels for the future. The aim of this work was to generatetransgenic Chlorella strains which are able to produce H2 by solar-driven pathway thatelectrons from photosynthetic electron transport chain can be transferred to hydrogenase.Contemporaneously, foreign hydrogenases will be overexpressed in transgenic Chlorella topromote H2 productionFirst year project: Photostimulation of H2 production in the transgenic Chlorellaupon suppression of its O2-evolving system by applying RNA interference (RNAi). A setof short interference RNAs (siRNAs) will be introduced to Chlorella to interfere the geneproducts of oxygen-evolving complex (OEC) subunits and depleting O2 evolution: OEC ofPSII is the major site for O2 evolution. If the genes encoding psbO, psbP or psbQ proteins ofOEC are silenced, then the O2 evolution should be inhibited. Therefore, the designatedsiRNAs to interfere these proteins will be also applied to Chlorella and to createphysiologically anaerobiosis and induce hydrogenase (Hyd) expression.Second year project: Genetic engineering hydrogenase-overexpressed Chlorella. Itwas known that Hyd's from some bacteria were less sensitive to O2 as compared to those ingreen algae. Hence, cloning of O2-tolerant hydrogenase from bacteria would be carried out.Subsequently, overexpression of O2-tolerant bacterial hydrogenase into O2-depletingtransgenic Chlorella shall greatly promote H2 production.Third year project: Processing small-scale bioreactor for H2 production in geneticengineering Chlorella.We intend to set up a small-scale bioreactor for Chlorella algalcultures. Under different nutrients, illumination, or temperature conditions, an optimalcultivation will be subject to transgenic Chlorella. A facility for continuously collectionmeans of H2 gas will be employed.

現今化石燃料的使用不但造成環境污染,造成溫室氣體CO2 的大量排放,且面臨開採枯竭,因有開發zero-CO2 之其他能源之急切性。而「氫氣(H2)」就是理想的再生能源。藻類可利用太陽能,在特定之代謝模式下產生「H2」。不過,負責製造「H2」的產氫酶(hydrogenase, Hyd)必須在缺硫、無氧的狀態時才有活性,所以只能觀察到短期少量之H2 產生。因此本研究計畫欲建構單細胞綠藻小球藻Chlorella 為zero-CO2 光生化產H2 之綠色工場︰建構綠藻去O2 突變株,抑制放氧活性,同時表現同源Hyd 或轉殖外來的氧耐受性高的Hyd,如此小球藻既可利用太陽能量進行光反應,又能同時產「H2」。第一年計劃是利用RNA interference(RNAi)的方法干擾葉綠體中光系統II(photosystem II, PSII)的之放氧複合體(oxygen-evolving complex, OEC)次單元,例如psbO之蛋白表現。若是沒有OEC,水分子無法分解成O2。在無氧環境下,Hyd 的基因(hyd)會被誘導表現Hyd,可利用由光系統I (PSI)所獲得的電子,將氫質子還原成「H2」。第二個年計劃是轉殖外來氧耐受性高的Hyd。一般綠藻的Hyd 對O2 很敏感,但是有些非紫硫菌的Hyd 對O2 耐受性高,若是將其轉殖於無氧狀態的Chlorella,應能大量提升產氫效率。第三個年計劃是將篩選的Chlorella 轉殖株,以小型反應器培養,連接氣體收集器,測試不同培養基、光照、溫度等條件,找出Chlorella 最佳產生「H2」之培養條件。
其他識別: NSC99-2221-E005-074
Appears in Collections:生命科學系所

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