| Project/Area Number |
13660336
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
生物資源科学
|
|---|
| Research Institution | Fukuyama University |
|---|
Principal Investigator |
HATANO Takushi Fukuyama University, Fac.Life Science and Biotechnology, Dept.Biotechnology, Professor, 生命工学部, 教授 (60198752)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
MATSUZAKI Hiroaki Fukuyama University, Fac.Life Science and Biotechnology, Dept.Biotechnology, Associate Professor, 生命科学部, 助教授 (90222299)
|
|---|
| Project Period (FY) |
2001 – 2002
|
| Project Status |
Completed (Fiscal Year 2002)
|
| Budget Amount *help |
¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 2002: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2001: ¥2,700,000 (Direct Cost: ¥2,700,000)
|
|---|
| Keywords | yeast / triacylglycerol / secretive production / palmitoleic acid / triacylglycerol binding protein / 菌体外生産 / 油脂分泌 / 脂質結合タンパク質 / Trichosporon sp. / Saccharomyces cerevislae |
|---|
| Research Abstract |
This research was performed to realize the secretive production system of useful microbial lipids.
1) Two types of the mutant strains, E228 and L-12, were obtained from the parent yeast Trichosporon sp. strain SH45Y. The mutant accumulated triacylglycerols (TG) in extracellular space, respectively. The strain E228 secreted TG containing palmitoleic acid on fatty acid substrates (2.4g/I/3 days), on the other hand the mutant L-12 excreted over 75% of total lipids on glucose.
2) It was made clear that the strain L-12 possesses the TG-transport system across the cell membrane. This mutant produced several species of TG-binding proteins (45kDa, 30kDa and 20kDa) in intra- and extracellular spaces. We determined N-terminal amino acid sequences of the 45kDa and 20kDa proteins. It was suggested that the gradients of affinities of these TG-binding proteins to TGs generate the motive forces of TG transport.
3) One mutant strain STG1 which secretes TGs (6% of total) was isolated from the laboratory yeast Saccharomyces cerevisiae. On the tetrad analysis, the phenotype of this mutant was complemented by wild type gene and derived from one gene mutation. This mutant strain, therefore, is thought a good tool for investigation of molecular mechanism of TG secretion in yeast. Although molecular cloning of wild type gene has tried vigorously, cell aggregation of this mutant was found a great barrier for gene cloning. The experiments to reduce cell aggregation are in progress.
|
