成果報告書詳細
管理番号20130000000192
タイトル*平成24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)微細藻類の改良による高速培養と藻体濃縮の一体化方法の研究開発
公開日2013/6/22
報告書年度2012 - 2012
委託先名株式会社IHI 国立大学法人神戸大学 株式会社ネオ・モルガン研究所
プロジェクト番号P10010
部署名新エネルギー部
和文要約ラボスケールにおいて、光合成のみで、安価に大量培養可能なことがわかっている「高速増殖型ボツリオコッカス」を使用して、人工光や糖源等のエネルギーを使わず、純粋な太陽エネルギーのみで、かつ専用リアクターなども使わない低コスト・低エネルギーの培養条件において、藻体生産性、藻油含有率、藻油回収率に優れた培養・回収プロセスを最終的に1000L規模の培養スケールにおいて実現することを目標とする。さらに、屋外での事業化に資する大規模培養の実現やオイル生産性のさらなる向上には、育種による改良が必須である。そこで、不均衡変異導入法による高速増殖型ボツリオコッカスのさらなる育種・改良により、コロニーサイズの拡大した改良株、浮上特性が向上した改良株、多糖分泌が低減された改良株を個別に獲得し、さらにこれらの特性を1つの株に集約すること(多重変異株の獲得)を最終目的とする。加えて、将来の遺伝子組換え技術による育種のために、遺伝子銃を用いたボツリオコッカス核ゲノムへの高効率で安定的な遺伝子導入技術と単細胞からの藻体再生技術を含む一連の遺伝子組換えによるボツリオコッカス形質転換系の開発を継続して行っている。
英文要約 Botryococcus braunii is known as a fast growing and high oil producing strain. The hyper-growth Botryococcus strain that can be cultivated massively by only photosynthetic energy had been found in our lab experiments. The eventual objective of this project is to establish the integrated oil production process, consisting of outdoor cultivation in 1000L scale using only sunlight as light source, harvesting, concentrating, and oil extraction in low cost using the hyper-growth Botryococcus. To achieve this objective following developments has been carried on. (i) To verify outdoor growth abilities of the hyper-growth Botryococcus, several tens to hundreds litter-scale outdoor cultivations using only sunlight were examined. Under weak sunlight during winter, only a little growth of the hyper-growth Botryococcus was observed. Preliminary, growth experiments were performed in an area with stronger sunlight, which results in prospect of target growth rate. Under any sunlight conditions, crude-oil contents of the hyper-growth Botryococcus maintain more than 40%. Further expansion of cultivation scale is going to be tried in the later stage. (ii) To establish an integrated process of harvesting and cultivation, the hyper-growth Botryococcus is needed to be improved. The natures such as larger colony size, quick buoyant ability after stopping agitation, and low secretion of bi-product, are considered to be advantageous for harvesting and cultivation. To acquire phenotypes possessing these natures, mutation was induced using disparity mutagenesis technology, and 200,000 of mutant library were established. After selective cultivation of mutants from the library, each phenotype was acquired. Acquisition of mutant having combined three natures described above is going to be examined in the later stage. (iii) For further improvement of the hyper-growth Botryococcus in the future, genetic recombination tools that will achieve stable gene expression has been developed. A plasmid vector including modified gene, antibiotic resistant gene and reporter gene was constructed. Gene introduction method with a particle gun has been examined using this plasmid. The introduction of the gene is being examined by selective cultivation. Stable gene expression is going to be examined by PCR detection of the target gene and microscopic observation of fluorescent protein expression in the hyper-growth Botryococcus’ cells. In the coming fiscal year the outdoor growth ability of improved Botryococcus will be examined in 1000 litter scale.
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