Study on molecular mechanisms of uptake, conversion, and excretion of lipids in yeast.
酵母脂质摄取、转化和排泄的分子机制研究。
基本信息
- 批准号:14206010
- 负责人:
- 金额:$ 34.36万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (A)
- 财政年份:2002
- 资助国家:日本
- 起止时间:2002 至 2004
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
To develop yeast systems for production of useful chemicals from alkanes and fatty acids, it is necessary to study how these natural resources are utilized by yeasts. In this project, we analyzed two yeast species that can utilize n-alkanes and fatty acids as carbon sources, Yarrowia lipolytica and Candida maltosa. Y.lipolytica has a haploid genome of which sequence analysis was completed recently. C.maltosa has a diploid genome and has been used for production of long chain dicarboxylic acids (DCA). We obtained following new results.1.Y.lipolytica takes up n-hexadecane in an energy dependent manner. This uptake is induced by alkane and repressed by glycerol and by mutations in peroxin genes.2.The production of long chain DCA by C.maltosa was improved by the increased expression of CmCDR1 gene that encodes an ABC transporter.3.We identified an alkane-responsive element ARE1 on the promoter of alkane-inducible genes. ARE1 dependent lacZ reporter expression system was constructed and integrated into the Y.lipolytica genome. By mutagenizing the resultant recombinant strain, we obtained one mutant defective in YAS1 gene. The yas1 mutant and Δyas1 did not grow on n-decane but grow on oleic acid. Yas1p is localized in nucleus and has a basic helix loop helix (bHLH) motif. CHIP assay proved that Yas1p-HA bound the ALK1 promoter region. From these results, we concluded that Yas1p is a transcription factor for n-alkane inducible gene expression. Since Yas1p bHLH region has homology to Ino4p bHLH of S.cerevisiae and Ino4p forms heterodimer with Ino2p, another protein with bHLH motif, we identified a gene YAS2 by searching Y.lipolytica genome sequence data and proved that Yas2p binds to Yas1p and is responsible for the alkane-responsible gene expression.
为了开发酵母系统,从烷烃和脂肪酸中生产有用的化学物质,有必要研究酵母如何利用这些自然资源。在本项目中,我们分析了两种能够利用正构烷烃和脂肪酸作为碳源的酵母菌:解脂亚罗维菌和麦芽假丝酵母。溶脂耶尔森菌具有单倍体基因组,其序列分析已于近期完成。麦芽单胞菌具有二倍体基因组,已被用于生产长链二元酸(DCA)。我们得到了以下新的结果:1.解脂酵母以能量依赖的方式摄取正十六烷。这种摄取是由烷烃诱导的,并被甘油和过氧化物素基因突变所抑制。2.通过增加编码ABC转运蛋白的CmCDR1基因的表达,麦芽假单胞菌合成长链DCA的能力得到了提高。3.我们在烷烃诱导基因的启动子上发现了一个烷烃反应元件ARE 1。构建了依赖are1的LacZ报告基因表达系统,并整合到溶脂耶尔森菌基因组中。对得到的重组菌株进行诱变,获得一株YAS1基因缺陷突变株。YAS1突变体和ΔYAS1不能生长在正庚烷上,而能生长在油酸上。Yas1p定位于细胞核,具有一个基本的螺旋环状螺旋(BHLH)基序。芯片实验证明Yas1p-HA结合了ALK1启动子区域。这些结果表明,Yas1p是一个正构烷烃诱导基因表达的转录因子。由于Yas1p的bHLH区与酿酒酵母的Ino4p有同源性,而Ino4p与另一种含有bHLH基序的蛋白质Ino2p形成异源二聚体,因此我们通过对解脂耶尔森氏菌基因组序列的检索,确定了Yas2基因,并证明Yas2p与Yas1p结合,负责烷烃相关基因的表达。
项目成果
期刊论文数量(38)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Peroxisome deficiency represses the expression of n-alkane-inducible YIALK1 encoding cytochrome P450ALK1 in Yarrowia lipolytica
- DOI:10.1016/s0378-1097(02)00838-8
- 发表时间:2002-08-27
- 期刊:
- 影响因子:2.1
- 作者:Sumita, T;Iida, T;Ohta, A
- 通讯作者:Ohta, A
Pseudohyphal growth in a dimorphic yeast, Candida maltosa, after disruption of C-GCN4 Gene, a homolog of Saccharomyces cerevisiae GCN4.
C-GCN4 基因(酿酒酵母 GCN4 的同源物)被破坏后,二态性酵母(麦芽糖假丝酵母)中的假菌丝生长。
- DOI:
- 发表时间:2002
- 期刊:
- 影响因子:0
- 作者:H.Takaku;H.Horiuchi;M.Takagi;A.Ohta
- 通讯作者:A.Ohta
The dual functions of biphenyl-degrading ability of Pseudomonas sp.KKS102 : energy acquisition and substrate detoxification.
假单胞菌KKS102的联苯降解能力的双重功能:能量获取和底物解毒。
- DOI:
- 发表时间:2003
- 期刊:
- 影响因子:0
- 作者:M.Delawary;Y.Ohtsubo;A.Ohta
- 通讯作者:A.Ohta
H.-S.Lee, K.Miyauchi, et al.: "Employment of human estrogen receptor β ligand-binding domain and co-activator SRC1 nuclear receptor binding domain for the construction of a yeast two hybrid detection system for endocrine disrupters"J.Biochem.. 131. 399-40
H.-S.Lee、K.Miyauchi 等人:“利用人雌激素受体 β 配体结合域和共激活剂 SRC1 核受体结合域构建酵母两种内分泌干扰物混合检测系统”J .生物化学.. 131. 399-40
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
A.Makino, T.Baba, et al.: "Cinnamycin (Ro 09-0198) Promotes Cell Binding and Toxicity by Inducing Transbilayer Lipid Movement"J.Biol.Chem.. 277(in press). (2002)
A.Makino、T.Baba 等人:“肉桂霉素 (Ro 09-0198) 通过诱导跨双层脂质运动促进细胞结合和毒性”J.Biol.Chem.. 277(印刷中)。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
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OHTA Akinori其他文献
OHTA Akinori的其他文献
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{{ truncateString('OHTA Akinori', 18)}}的其他基金
Study on lipid metabolism in the industrial yeast Yarrowia lipolytica
工业酵母解脂耶氏酵母脂质代谢研究
- 批准号:
24380045 - 财政年份:2012
- 资助金额:
$ 34.36万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
System biologic approach to lipid traffic and conversion in eukaryotic microorganisms and its application
真核微生物脂质运输和转化的系统生物学方法及其应用
- 批准号:
17208008 - 财政年份:2005
- 资助金额:
$ 34.36万 - 项目类别:
Grant-in-Aid for Scientific Research (A)
Improvement of long chain dicarboxylic acid production by fortifying omega-oxidation, excretion of products and lipase production in yeast
通过强化酵母中的欧米伽氧化、产物排泄和脂肪酶生产来改善长链二羧酸的生产
- 批准号:
12556011 - 财政年份:2000
- 资助金额:
$ 34.36万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Mechanism-Based Biodegradability Prediction of Dehalogenase
基于机制的脱卤酶生物降解性预测
- 批准号:
10044197 - 财政年份:1998
- 资助金额:
$ 34.36万 - 项目类别:
Grant-in-Aid for Scientific Research (B).
Analysis on the mechanism of ER membrane proliferation upon ER protein overproduction in yeast
酵母内质网蛋白过量产生内质网膜增殖机制分析
- 批准号:
10460034 - 财政年份:1998
- 资助金额:
$ 34.36万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Construction of terminal oxidation systems for hydrocabon chain A use of a variety of yeast cytochrome P450 genes
多种酵母细胞色素P450基因的烃链末端氧化系统的构建
- 批准号:
10556015 - 财政年份:1998
- 资助金额:
$ 34.36万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Breeding of a flocculent, highly alcohol-producing yeast
絮状高产酒精酵母的育种
- 批准号:
08556012 - 财政年份:1996
- 资助金额:
$ 34.36万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Study on the development and differentiation of endoplasmic reticulum of yeast from the view point of lipid synthesis.
从脂质合成的角度研究酵母内质网的发育和分化。
- 批准号:
07456045 - 财政年份:1995
- 资助金额:
$ 34.36万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Genetic Engineering of Long Chain Dicarboxylic Acid Production in Yeast
酵母生产长链二元酸的基因工程
- 批准号:
06556013 - 财政年份:1994
- 资助金额:
$ 34.36万 - 项目类别:
Grant-in-Aid for Developmental Scientific Research (B)
Role of Amino-phospholipid Synthesis in Biogenesis of Endoplasmic Reticulum of Yeast
氨基磷脂合成在酵母内质网生物合成中的作用
- 批准号:
04660080 - 财政年份:1992
- 资助金额:
$ 34.36万 - 项目类别:
Grant-in-Aid for General Scientific Research (C)
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