成果報告書詳細
管理番号20140000000270
タイトル*平成25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) 海洋性緑藻による油脂生産技術の研究開発
公開日2014/11/19
報告書年度2013 - 2013
委託先名国立大学法人神戸大学 大学共同利用機関法人自然科学研究機構基礎生物学研究所 DIC株式会社
プロジェクト番号P10010
部署名新エネルギー部
和文要約
英文要約Title: Development of Oil Production Process Using Marine Green Algae (FY2012-FY2015) FY2013 Annual Report

A. Kobe University
(1) Development of lipid producing system
In cultivation of strain JSC4, various media were selected, and then several stresses were explored to optimize the lipid productivity. Under the best condition, the high biomass productivity (539 g/m3/day) were obtained. The lipid content was promoted from 53.5% (2% sea salt) to 64.0% (5% sea salt) with an increase in sea salt; however, the lipid productivity under high salinity was decreased, suggesting that it could inhibit the cell growth but trigger the lipid accumulation. Hence, the newly gradient-salinity operation system was constructed to increase lipid accumulation with the satisfactory biomass productivity. The highest lipid productivity (223 g/m3/day) with high lipid content (59.4%) was obtained.
(2) Metabolic analysis of JSC4
After metabolite extraction and CE/MS analyzing, it showed that there were positive correlations between lipid synthesis and several metabolites, indicated these major metabolites could be used as markers for lipid production. Hereafter, further enhancement of lipid productivity was expected by improving the bottle-neck reaction of metabolic flow via genetic engineering.

B. National Institute of Basic Biology
(1) Genetic transformation of strain JSC4
The strain used in this study was identified as Chlamydomonas sp. JSC4 by 18S rRNA. A vector pRT-GenD-CrmVenus-aphVIII was developed and introduced into strain JSC4 cells by a multipulse electropolator. Fifty colonies per plate were obtained on average. The genomic DNAs prepared from these colonies contained aphVIII gene encoding an enzyme capable of inactivating paromomycin, confirmed by a PCR analysis.
(2) Analysis of factors limiting the photosynthetic activity of strain JSC4
Strain JSC4 was grown continuously in an open pond of DIC Corporation with monitoring several parameters such as light, temperature, photosynthetic quantum yield, and so on, indicated the cells were suffered from an excess light immediately and the photosynthetic quantum yield were low during the day because of the excess light, which limited the overall photosynthetic rate.

C. DIC Corporation
(1) Development of outdoor mass cultivation technology
Outdoor experiments have been demonstrated by using 1.2 and 2.4 m2 raceway ponds set-up at Earthrise Nutritionals LLC, USA. Firstly, outdoor cultivation of strain JSC4 was difficult because of the contamination of parasitic fungus. After investigations, the contamination was controlled by adding sea salt, and a monthly semi-continuous cultivation was achieved. The maximum biomass productivity was 22 g/m2/day.
(2) System evaluation and preliminary cost estimation
Preliminary oil production cost estimation is 292 yen/L, based on Vasudevan et al (2012), and under the assumption that 400 hectare cultivation facility, 40 g/m3/day lipid productivity with 60% oil content.
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