成果報告書詳細
管理番号20140000000218
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) 油糧微生物ラビリンチュラを利用したジェット燃料・船舶燃料生産の研究開発
公開日2014/5/17
報告書年度2013 - 2013
委託先名株式会社Biomaterial in Tokyo 国立大学法人宮崎大学
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 Bio-Jet Fuel and Marine Fuel Production System using thraustochytrids (FY2013-FY2014) FY2013 Annual Report

Details and Results of the Research
1.Creation of Labyrinthula Strains that Utilize Xylose and Arabinose
i. Selection of Labyrinthula Strains Capable of Utilizing for Saccharified Biomass Solution (done by University of Miyazaki)
Through a screening process, the following strains of Labyrinthulas that are capable of utilizing the C5 sugars (ie: Arabinose and Xylose) in saccharified biomass solutions were isolated.
-20 strains that utilize xylose when it is the sole carbon source in the chemically defined medium.
-21 strains that utilize arabinose when it is the sole carbon source in the chemically defined medium.
From the above-mentioned 41 strains, 7 strains were tested for antibiotic resistance to Zeocin, Blasticizin, Hygromycin, and G418. All strains showed sensitivity to Blasticizin and G418, so their resistance will be used as a selectable marker for the genetic transformation process. The mh1918 strain also showed sensitivity to Zeocin, allowing for a third marker to be used.
ii.Labyrinthula Strains Optimized for Growth on Saccharified Biomass
(done by BITS)
In preparation for genetic transformation of the selected strains, codons responsible for C5 sugar uptake and metabolism (xylose reductase (XR), xylitol dehydrogenase (XDH), xylose kinase (XKS1)) were optimized to ensure maximum expression by the modified strains. Preparation of expression cassettes with the target genes and the codons are now underway.

2.Evaluation of Cultivation Conditions to Maximize Lipid Production
i.Development of High-Density Cultivation Techniques Using Saccharified Biomass (done by University of Miyazaki)
Small-scale cultivation was done using the above-mentioned 41 selected strains. Each strain was cultivated in 24-well microplates containing chemically defined media with either xylose or arabinose as the sole carbon source. A procedure for analyzing C5 sugar concentration using ion exchange chromatography has been developed, and will be used in trials in the upcoming year to determine C5 sugar consumption.

3.Evaluation of Labyrinthula Oils for Jet Fuel Use (done by Cosmo Oil)
Oils produced by Labyrinthula will be extracted for analysis to determine its composition and compatibility for jet fuel. Extraction and refining methods such as “cracking” will also be evaluated on an industrial scale to determine its feasibility.
-Composition Analysis of Labyrinthula Oil
An analysis of Labyrinthula oil was done using gas chromatography and mass spectroscopy to determine its distillation curve. The same tests were done with palm oil, fatty acid methyl esters and triglycerides of stearic acid and oleic acid (both found in Labyrinthula oils) and a comparison is underway with the results. Once the Labyrinthula strain for large-scale cultivation has been decided, the process will be repeated for comparison.
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