成果報告書詳細
管理番号20110000000842
タイトル*平成22年度中間年報 バイオマスエネルギー技術研究開発 戦略的次世代バイオマスエネルギー利用技術開発事業(次世代技術開発) 遺伝子改良型海産珪藻による有用バイオ燃料生産技術開発
公開日2011/6/7
報告書年度2010 - 2010
委託先名国立大学法人高知大学 国立大学法人京都大学 国立大学法人東京大学大学院農学生命科学研究科 株式会社ユーグレナ
プロジェクト番号P10010
部署名新エネルギー部
和文要約和文要約等以下本編抜粋:1. 研究開発の内容及び成果等
無尽蔵とも言える海水を利用して増殖可能な海産珪藻Rhizosolenia属等を研究開発対象とし、新たに開発した遺伝子操作技術を用いることにより、液体燃料として有用な炭化水素を高度に生産する能力を備えた形質転換海産微細藻類株の創生を目指す。同時に、これらの海産珪藻を用いて、太陽光を有効に利用しながら高効率に培養可能な、バイオリアクタならびにその最適運転条件を解明・確立することにより、バイオ燃料の生産性を向上させる。また、コストのかからない炭化水素抽出法を開発するために、炭化水素生産株を溶藻するウイルス分離を目指す。同時に、バイオ燃料抽出後の藻体残渣から、副産物としてバイオエタノールなどを回収することに加えて、珪藻土等の高付加価値を有する物質を獲得することにより、2030年の実用化に向けて、コストに見合うバイオ燃料生産の実現を目指す。以下に、平成22年度に得られた研究成果について纏めて示す。
英文要約Title: Development of a Biofuel Production System using Genetic Engineered Marine Diatoms (FY2010-FY2011) FY2010 Annual Report
The objective for this project is to develop a biofuel production system using the genetic engineered marine diatoms which produce hydrocarbons. The project consists of the four subthemes. One of the purposes of the first subtheme is to isolate marine diatom strains which contain large amounts of lipids, especially hydrocarbons. For this, we have isolated about one hundred strains of Rhizosolenia spp. from Japanese coastal waters. In order to develop a method for rapid screening of cellular lipid content in diatoms, a Nile red in situ staining method was investigated. We clarified that 5% DMSO (dimethyl sulfoxide) treatment increased Nile red permeability of the cell membranes without cell disruption. Another purpose of the first subtheme is to develop a stable transformation system of the marine diatoms essential for their genetic engineering. For this, we have succeeded in construction of a new transformation vector containing a new viral promoter isolated from a virus infecting a marine diatom. Antibiotics sensitivities of Rhizosolenia setigera CCMP1694 which produces hydrocarbons were also clarified. The purpose of the second subtheme is to characterize and quantify useful compounds and to isolate genes concerned with hydrocarbon production in marine diatoms. Analyses of R. setigera CCMP1694 hydrocarbons by GC-MS and 1H-NMR indicated these hydrocarbons were rhizenes, triterpenoids with molecular formula of C30H52 or C30H50. RNA was isolated from R. setigera to create a cDNA library. R. setigera CCMP1694 also contains other useful compounds such as ~-carotene as its major carotenoid. The purpose of the third subtheme is an improvement of growth rate, and content ratio of neutral lipids and hydrocarbons in the marine diatom Rhizosolenia. Culture experiments for elucidation of the optimum growth conditions clarified that the addition of sodium bicarbonate to SWM3 medium enhanced growth of R. setigera. The original cells of R. setigera were not able to grow above 30°C using SWM3 medium. After passage culture, the adapted cells of R. setigera were able to grow at 32°C. Moreover, bioreactor systems suitable for R. setigera were discussed. In the fourth subtheme, we studied the necessary parameters to compare the productivity of hydrocarbons by microalgae. In addition, assuming the amounts of hydrocarbons from the previous studies, we also examined the processing methods. We compiled the parameters necessary for evaluation of lipids as fuel but the methods of refinement are still in progress. In the coming fiscal year, we are aiming to evaluate the cost of culture methods including the extraction and refinement processes of the lipids for industrial use.
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