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Large scale collection and their industrial application of hyperthemophilic archaeal genome from deep sea hydrothermal envirenments

深海热液环境超嗜热古菌基因组的大规模采集及其工业应用

基本信息

批准号：
16208020
负责人：
SAKO Yoshihiko
金额：
$ 30.62万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2007
项目状态：
已结题

项目摘要

In this study, we first investigated the deep sea hydrothermal vent environments and its microbial community. Then we collected the large scale genome information of hyperthermophilic microrganisms distributed in those hot vents and tried to develop their genetic resources to industrial application of hyperthermophilies.(1) To determine whether the geochemical process (boiling/distillation) of subseafloor hydrothermal fluids affects the formation of microbial communities, we examined the microbial communities at three different vent sites located within a few tens meters of one another in the Iheya North field. In addition to chimney structures, colonization devices capturing subseafloor communities entrained by the vent fluids were studied, using culture-dependent and-independent methods. Microbial analyses demonstrated the occurrence of distinctive microbial communities in each of the hydrothermal niches. Within a chimney structure, there was a transition from a mixed community of me … More w/Miles and thermophiles in the exterior parts to thermophiles in the interior In comparison to gas-depleted and normal fluid% gas-enriched fluids harbored more abundant chemolithoautotrophs with gaseous component-dependent energy metabolism, such as hydrogenotrophic methanogenesis. Subseafloor phase-separation and -segregation may play a key role in supplying energy and carbon sources to vent-associated chemolithoautotrophs and subvent microbial communities.(2) In the Iheya North field in the Mid-Okinawa Dough we studied the spatial distribution, diversity and physiological characteristics of the epsilon-Proteobacteria in various microbial habitats in the vicinity of this deep-sea hydrothermal vent The 111 gill samples were predominantly colonized by epsilon-Proteobacteria Energy metabolism of epsilon-Proteobacteria isolates was highly versatile. Tree topology generated from the metabolic traits was significantly different from that of 16S rRNA tree. Culturability of epsilon-Proteobacteria in various microbial habitats differed among the phylogenetic subgroups. Members of Sulfurimonas were characterized by the robust culturability, and the other phylogenetic subgroups appeared to lose culturability in seawater, probably because of the sensitivity to oxygen. These results provide new insight into the ecophysiological characteristics of the deep-sea hydrothermal vent epsilon-Proteobacteria, which has never been assessed by comparative analysis of the 16S rRNA genes.3) Over 50 introns have been reported in archaeal rRNA genes (rDNAs), a subset of which nests putative homing endonuclease genes. We studied the identification and characterization of a novel archaeal LAGLIDADG-type homing endonuclease, I-ApeI, encoded by the ApeK1.S908 intron within the 16S rDNA of Aeropyrum pernix K1. I-ApeI consisits of 222 amino acids and harbors two LAGLIDADG-like sequences. It recognizes the 20 bp non-palindromic sequence 5'-GCAAGGCTGAAAC ↓ TTAAAGG and cleaves target DNA to produce protruding tetranucleotide 3' ends. Either Mn^<2+> or Co^<2+> can be substitude for Mg^<2+> as a cofactor in the cleavage reaction. Of the 20 bases within the minimal recognition site, 7 are essential for cleavage and are located at positions proximal to the cleavage sites. Less

在本研究中，我们首先调查了深海热液喷口环境及其微生物群落。同时，我们还收集了分布于这些热泉中的超嗜热微生物的大规模基因组信息，并试图将其遗传资源开发为超嗜热微生物的工业应用。(1)为了确定海底下热液流体的地球化学过程（沸腾/蒸馏）是否影响微生物群落的形成，我们研究了位于Iheya North油田彼此相距几十米的三个不同喷口处的微生物群落。除了烟囱结构，殖民化装置捕获海底下的喷口流体夹带的社区进行了研究，使用文化依赖和独立的方法。微生物分析表明，在每一个热液壁龛独特的微生物群落的发生。在一个烟囱结构中，我从一个混合的社区过渡到一个混合的社区， ...更多信息与贫气和正常流体相比，富气流体具有更丰富的化能无机自养生物，其具有依赖于气体组分的能量代谢，例如氢营养甲烷生成。海底相分离和分离可能发挥关键作用，在提供能源和碳源喷口相关的化能无机自养生物和subvent微生物群落。(2)在冲绳中部的Iheya北场，我们研究了在这个深海热液喷口附近的各种微生物栖息地的ε-变形菌的空间分布，多样性和生理特性。111个鳃样品主要由ε-变形菌定殖。ε-变形菌分离物的能量代谢是高度通用的。由代谢性状生成的树状图与16 SrRNA树状图有显著差异。ε-变形菌在不同微生物生境中的可培养性在系统发育亚群之间存在差异。硫酸单胞菌属的成员的特点是强大的可培养性，和其他系统发育的亚群似乎失去了在海水中的可培养性，可能是因为对氧气的敏感性。这些结果为深海热液喷口epsilon-Proteobacteria的生理生态学特征提供了新的见解，这是从未通过16 S rRNA基因的比较分析进行评估的。3）在古细菌rRNA基因（rDNA）中已经报道了50多个内含子，其中一个子集嵌套了推定的归巢核酸内切酶基因。我们研究了一种新的古细菌LAGLIDADG型归巢核酸内切酶，I-ApeI，编码的ApeK1.S908内含子内的16 S rDNA的Aeropyrum pernix K1的鉴定和表征。I-ApeI由222个氨基酸组成，具有两个LAGLIDADG样序列。它识别20 bp非回文序列5 '-GCAAGGCTGAAAC ↓ TTAAAGG，并切割靶DNA以产生突出的四核苷酸3'末端。Mn^<2+>或Co^<2+>都可以取代Mg^<2+>作为裂解反应中的辅助因子。在最小识别位点内的20个碱基中，7个是切割所必需的，并且位于切割位点附近的位置。少