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Molecular and cellular biology and development of useful cellular functions of microorganisms which can utilize future-type natural resources

分子和细胞生物学以及可利用未来型自然资源的微生物有用细胞功能的开发

基本信息

批准号：
13854008
负责人：
SASAKI Yasuyoshi
金额：
$ 78.87万
依托单位：
Graduate School of Agriculture, Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (S)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2005
项目状态：
已结题

项目摘要

Among future-type natural resources, we focused on utilization of natural gas-derived-C1-compounds (eg., methanol) and various type of alkanes (gaseous and isoalkanes), by various microorganisms. We tried to understand cellular functions of such microorganisms from three distinct aspects, i.e., biochemisty, cell biology, and molecular biology. Throughout the research, obtained information and reagents were considered for application to biotechnology. Our biochemical research has revealed novel metabolism for bacterial formadehyde-fixation and formaldehyde detoxification, iso-alkane and gaseous alkane oxidation at gene levels (J Bact, 2004, 2005 : FEMS Microbiol 2002). And also, novel detoxification systems against formaldehyde and reactive oxygen speicies were discovered with methanol-utilizing yeasts (JBC 2001 ; J. Bact 2001 ; Yeast 2004 ; Microbiology 2003). Our cellular and morphological studies on methanol-utilizing yeasts have revealed up to 30 molecular machineries for organelle … More (peroxisome) degradation, and identified a previously-unknown intracellular membrane dynamics, e.g., appearance of a novel membrane structure, micropexophagic membrane apparatus (MIPA), as well as novel vacuolar dynamics. These were further studied from the scope of molecular functions of newly-identified ATG gene products (EMBO J 2003 ; Mol Biol Cell 2004, 2005 ; Dev Cell 2003). These studies and obtained reagents were revealed to be useful for production of heterologous proteins by methylotrophic yeasts. Molecular biological research has focused on the regulatory expression of C1-enzymes in bacteria and yeasts. This study identified the first DNA-binding protein HxlR in Bacillus subtilis, which regulates formaldehyde-induced gene expression of formaldehyde-fixation pathway (Mol Microbiol 2005). And we also developed a novel formaldehyde- or formate-inducible promoter and valuable hosts with methanol-utilizing yeasts (Mol Gen Genet 2003). Using these reagents, we have succeeded in high level production of several useful proteins in an active form (Biosci Biotechnol Biochem 2002, 2003, 2004 ; Appl Microbiol Technol 2002.) Less

在未来型自然资源中，我们关注天然气衍生的C1化合物的利用（例如，甲醇）和各种类型的烷烃（气态和异烷烃）。我们试图从三个不同的方面来理解这些微生物的细胞功能，即，生物化学、细胞生物学和分子生物学。在整个研究过程中，考虑将获得的信息和试剂应用于生物技术。我们的生物化学研究揭示了细菌甲醛固定和甲醛解毒、异烷烃和气态烷烃氧化在基因水平上的新代谢（J Bact，2004，2005：FEMS Microbiol 2002）。此外，还发现了利用甲醇的酵母对甲醛和活性氧的新型解毒系统（JBC 2001 ; J. Bact 2001 ; Yeast 2004 ; Microbiology 2003）。我们对甲醇利用酵母的细胞和形态学研究揭示了多达30种细胞器的分子机制 ...更多信息（过氧化物酶体）降解，并确定了以前未知的细胞内膜动力学，例如，出现了一种新的膜结构，微胞吞膜装置（MIPA），以及新的空泡动力学。从新鉴定的ATG基因产物的分子功能范围进一步研究了这些（EMBO J 2003 ; Mol Biol Cell 2004，2005 ; Dev Cell 2003）。这些研究和获得的试剂被揭示为有用的生产异源蛋白的甲基营养型酵母。分子生物学研究集中在细菌和酵母中C1-酶的调控表达。本研究鉴定了枯草芽孢杆菌中的第一个DNA结合蛋白HxlR，其调节甲酰胺去固定途径的甲酰胺诱导的基因表达（Mol Microbiol 2005）。我们还开发了一种新的甲醛或甲酸诱导型启动子和利用甲醇的酵母的有价值的宿主（Mol Gen Genet 2003）。使用这些试剂，我们已经成功地以活性形式高水平生产了几种有用的蛋白质（Biosci Biotechnol Biochem 2002，2003，2004 ; Appl Microbiol Technol 2002）。少