成果報告書詳細
管理番号20150000000202
タイトル*平成26年度分中間年報 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2015/6/25
報告書年度2014 - 2014
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)
FY2014 Annual Report

A. Kobe University
(1) Development of oil producing technique
We found inappropriate irradiance inhibited the cell growth and oil production of Chlamydomonas sp. JSC4. The level of 3PGA and the rates of metabolic flux related to oil synthesis were increased under optimal light intensity, indicated appropriate light is vital to enhance oil production.
To consider the temperature shift outdoors, JSC4 was cultivated under 15-40°C. The highest biomass concentration of 3.5 g/L with lipid content of 50% was obtained under 30°C, which is 2-fold higher than that under 15°C and 40°C. To adapt low temperature, JSC4 was subculture under 15°C, resulting in higher growth.
Discovering a novel strain with high salt tolerance was vital to decrease the contamination risk outdoors. The mutated JSC4 with 7% salt tolerance was successfully constructed via heavy ion (12C5+) beam.
(2) Development of high oil producing strain
To improve lipid production, dynamic metabolic flux was performed to monitor the rate-limiting step of lipid synthesis in JSC4. The gene involved in the rate-limiting step was amplified from cDNA and then introduced into the vector of nuclear transgene. The mutant strain harboring the vector was successfully constructed.

B. National Institute of Basic Biology
(1) Improvement of transformation efficiency
We developed a high efficient transformation method in JSC4, which was 20 times higher.
(2) Analysis of factors limiting the photosynthesis of JSC4
We sequentially monitored photosynthetic parameters, light intensity and water temperature outdoors in Earthrise at summer and spring. The drastic reduction of photosynthesis during daytime was observed with a drop of growth in spring (The lowest night temp. = 10°C) compared with summer (The lowest night temp. > 20°C). The decrease of photosynthesis and growth rate in spring was recovered by keeping temperature above 20°C at night, indicated the lowest night temperature highly affects the photosynthetic efficiency during daytime.

C. DIC Corporation
(1) Optimization of outdoor mass culture
Outdoor cultivation of JSC4 was demonstrated by using 1.2, 2.4, and 50 m2 raceway ponds at Earthrise, USA. After investigation, we found appropriate salinity is able to control Chytrid without inhibiting JSC4 growth. Finally, semi-continuous cultivation in a 50 m2 raceway pond was successfully achieved. The biomass productivity was varied seasonally, from 8 to 20 g/m2/day. In addition, JSC4 was cultivated under saline stress in 100L flat-plate PBR, with a oil productivity of 49 g/m3/day. The optimization is under progress.
(2) Preliminary cost estimation
Preliminary oil cost estimation is 292 yen/L. This estimation was calculated under the assumption of 400 hectare of cultivation facility with 40 g/m3/day of oil productivity.
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