Kota, S.*; Rajpurohit, Y. S.*; Charaka, V. K.*; 佐藤 勝也; 鳴海 一成*; Misra, H, S.*
Extremophiles, 20(2), p.195 - 205, 2016/03
The multipartite genome of forms toroidal structure. It encodes topoisomerase IB and both the subunits of DNA gyrase (DrGyr) while lacks other bacterial topoisomerases. Recently, PprA a pleiotropic protein involved in radiation resistance in has been suggested for having roles in cell division and genome maintenance. In vivo interaction of PprA with topoisomerases has also been shown. DrGyr constituted from recombinant gyrase A and gyrase B subunits showed decatenation, relaxation and supercoiling activities. Wild type PprA stimulated DNA relaxation activity while inhibited supercoiling activity of DrGyr. Thus, we showed that DrGyr confers all three activities of bacterial type IIA family DNA topoisomerases, which are differentially regulated by PprA, highlighting the significant role of PprA in DrGyr activity regulation and genome maintenance in .
佐藤 勝也; 上田 涼史郎*; 長谷 純宏; 鳴海 一成*; 大野 豊
JAEA-Review 2015-022, JAEA Takasaki Annual Report 2014, P. 100, 2016/02
Bioremediation uses biological organisms to solve an environmental problem. However, microorganisms are sometimes not enough effective to achieve efficient recovery. Therefore, enhancing the functionality of microorganisms is needed to promote the active use of bioremediation. This study began with the purpose of development of Cs-accumulating bacteria by ion beam breeding technology. Previously, we suggested that is the most suitable species for the development of high Cs-accumulating bacteria by ion beam breeding technology. Candidates of Cs-accumulating mutant were screened from the mutagenized cells by ion beam breeding technology on the basis of the luminescent intensity and the cell area as indicators of intracellular Cs level and cell growth, respectively. As a result, we obtained 33 candidates, whose Cs-accumulating abilities were over 2-fold higher than that of the wild-type, from the population of mutagenized cells.
丸山 雄大*; 武田 喜代子*; 友岡 憲彦*; 佐藤 勝也; 大野 豊; 横山 正*
JAEA-Review 2015-022, JAEA Takasaki Annual Report 2014, P. 99, 2016/02
is one of wild legumes, which called as Hamaazuki, are distributed at the vicinity of the coast from tropical to sub-tropical regions and have high salt tolerance among terrestrial plants. can grow at 400 mM NaCl condition. Furthermore, the root nodules on have a symbiotic relationship with rhizobia that related to the genus having high stress tolerance (500 mM NaCl, 45 C, pH 10.5). Above both symbiotic partners showed extremely high salt tolerance, however, cannot develop root nodules with at over 80 mM NaCl conditions. To improve symbiotic performance of with under high salt conditions, bacterial cells were irradiated by carbon ion-beam at TIARA. However, so far, we could not obtain promising mutants having good symbiotic performance with Vigna marina under high salt conditions, yet.
荻野 千秋*; Kahar, P.*; Lee, J. M.*; 佐藤 勝也; 大野 豊; 近藤 昭彦*
JAEA-Review 2015-022, JAEA Takasaki Annual Report 2014, P. 105, 2016/02
Ethanol fermentation from hydrolyzed lignocellulosic biomass with yeast is very important. However, in common sense, there are several chemical inhibitors for yeast from hydrolyzed biomass. Therefore, screening of yeast strain, possessing the tolerance properties against these chemical inhibitors, should be found from nature resources or mutant libraries. To overcome the problem, we have screened yeast strains tolerant to many chemical inhibitors contained in bagasse sugar lysate (BSL) at high temperature (35 C) from 1500 strains out of national bio-resource center yeast collection. We found that F118 strain could grow and ferment BSL and produce ethanol. For increasing of the yield and ethanol fermentation efficiency of F118 strain, the adaptation of F118 strain to high temperature was conducted, followed by carbon ion beam irradiation. Consequently, we have successfully obtained F118S88 strain as a good candidate for ethanol production from BSL at high temperature.
上田 涼史郎*; 佐藤 勝也; 林 秀謙*; 鳴海 一成*; 大野 豊
JAEA-Review 2015-022, JAEA Takasaki Annual Report 2014, P. 101, 2016/02
It is suggesting a possibility that the polyphosphate granules in cells could accumulate harmful metal ions such as radiocesium to protect the cells from cytotoxicity. In this study, we investigated the accumulation of cesium and its association with intracellular polyphosphate in using the gene disruptant and overexpressing strains. The cesium accumulation level in the disruptant and overexpressing strains was compared with that of the wild-type strain. The cesium accumulations in the disruptant and overexpressing strains were slightly decreased and almost same, respectively. Like the intracellular polyphosphate level, the cesium accumulation level was significantly increased in the disruptant strain. These results suggested that the intracellular polyphosphate level was positively correlated with the cesium accumulation level in .
増渕 隆*; 渡口 和樹*; 林 秀謙*; 池永 裕*; 佐藤 勝也; 大野 豊
JAEA-Review 2015-022, JAEA Takasaki Annual Report 2014, P. 106, 2016/02
We conducted the gene function analysis of the high ethyl caproate producing sake yeast mutant (No.227) generated by ion beam breeding in order to develop a new method for sake yeast screening and find factors contributing the reduced ability to alcohol fermentation. The genome sequences of the No.227 and Kyokai 901 (its parental strain) were determined by a whole-genome shotgun strategy using pyrosequencing method. And the determined sequences were compared with that of the sake yeast strain Kyokai 7, which is characterized by the fermentation properties. Consequently, the No.227 carried mutations in the and genes as heterozygous state. It was suggested that these mutations in the and genes in the No.227 might be attributed to the high ethyl caproate production and the low alcohol fermentation ability, respectively.
佐藤 勝也; 小野寺 威文*; 面曽 宏太*; 武田-矢野 喜代子*; 片山 豪*; 大野 豊; 鳴海 一成*
Genome Announcements (Internet), 4(1), p.e01631-15_1 - e01631-15_2, 2016/01
was isolated as a gram-negative, red-pigmented, radioresistant, rod-shaped bacterium from freshwater fish. The draft genome sequence of ATCC 43672 was 4,092,497 bp, with an average G+C content of 67.5% and comprised of 4 circular contigs and 3 linear contigs that was deposited at DDBJ/EMBL/Genbank under the accession number BCMS00000000. The draft genome sequence indicates that possesses a DNA damage response regulator (encoded by homolog) and radiation-desiccation response regulons (, , , and homologs etc.), which are involved in the unique radiation-desiccation response system in . As it is for , seems to employ the same radioresistant mechanisms. In future, the draft genome sequence of will be useful for elucidating the common principles of the radioresistance based on the extremely efficient DNA repair mechanisms in Deinococcus species by the comparative analysis of genomic sequences.
藤浪 俊*; 武田 喜代子*; 小野寺 威文*; 佐藤 勝也; 清水 哲*; 若林 佑*; 鳴海 一成*; 中村 顕*; 伊藤 政博*
Genome Announcements (Internet), 3(5), p.e01005-15_1 - e01005-15_2, 2015/09
sp. ME121 was isolated from soil as a mixed single colony with sp. 32K during our screening of L-glucose-utilizing microorganisms, and its growth was enhanced by coculture. It was expected that genomic analysis of this bacterium would provide novel information on coculture-dependent growth enhancement. The genomic information of symbiotic bacteria could be of use for studying the molecular mechanisms underlying microbial symbiosis. The draft genome sequence of sp. ME121 is 7,096,979 bp in total length and comprises 197 large contigs ( 500 bp) that was deposited at DDBJ/EMBL/GenBank under the accession number BBUX00000000. The draft genome sequence shows that sp. ME121 has some genes that encode putative methanol/ethanol family PQQ-dependent dehydrogenases involved in methylotrophy. Some unknown factor provided by the coculture may contribute to increase the growth of sp. ME121.
大野 豊; 長谷 純宏; 佐藤 勝也; 野澤 樹; 鳴海 一成*
放射線と産業, (138), p.17 - 20, 2015/06
佐藤 勝也; 上田 涼史郎; 長谷 純宏; 鳴海 一成*; 大野 豊
JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 117, 2015/03
Bioremediation uses biological organisms to solve an environmental problem. However, microorganisms are sometimes not enough effective to achieve efficient recovery. Therefore, enhancing the functionality of microorganisms is needed to promote the active use of bioremediation. This study began with the purpose of development of Cs-accumulating bacteria by ion beam breeding technology. We investigated which members are better suited for Cs-accumulation in the genus (, , , , and ) exhibit extraordinary radioresistant. The Cs concentration in the cells exhibited over 4-fold compared with that of the by atomic absorption spectrometer analysis. This result suggested that and are best two suitable species for the development of high Cs-accumulating bacteria by ion beam breeding technology.
上田 涼史郎; 佐藤 勝也; 林 秀謙*; 鳴海 一成*; 大野 豊
JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 118, 2015/03
It is suggesting a possibility that the polyphosphate granules in cells could accumulate harmful metal ions such as radiocesium to protect the cells from cytotoxicity. has been considered as a microorganism for bioremediation under highly radioactive contaminated environments. We generated the polyphosphate biosynthesis-related genes ( and ) disruptant strains and characterized their disruption effect to clarify the role of polyphosphate for the accumulation of cesium in . The intracellular levels of polyphosphates in the disruptant and overexpressing strains were almost same and slightly increased, respectively. On other hand, the intracellular level of polyphosphates significantly increased in the disruptant strains, suggesting that the gene plays an important role in the accumulation of polyphosphate in .
武田 喜代子*; 佐藤 勝也; 鳴海 一成*; 大野 豊; 大津 直子*; 横山 正*
JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 120, 2015/03
Major constraint of biofertilizer utilization in agricultural practice is a viability loss of beneficial microorganisms in biofertilizers by high temperature stress during both storage and transportation. Therefore, we have tried to improve high temperature tolerance of USDA110 using carbon-ion beams accelerated by an AVF cyclotron at TIARA. Consequently, we have obtained a high temperature tolerant mutant that can maintain a high survival rate at 42 C for at least 7 days in yeast-mannitol broth medium, and named it as M14. The genome sequence of M14 was determined by a whole-genome shotgun strategy. The acquirement of high temperature tolerance in M14 could be attributed to 18 base mutations and/or a large-scale inversion (1.27 Mb). In the future, we will conduct further analysis on these mutations in order to decipher the molecular mechanisms of high temperature tolerance of M14.
西東 力*; Fitriana, Y.*; 佐藤 勝也; 大野 豊; 鳴海 一成*
JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 121, 2015/03
Entomopathogenic fungi, such as , are economically important agents for integrated pest management programmes of insect pests. However, negative effects of temperature, such as heat stress that exceeds 35 C, can have serious deleterious effects on conidial germination and persistence, vegetative growth, sporulation, and/or the infection process in these entomopathogenic fungi. In this study, we attempted to enhance thermotolerance in a model entomopathogenic fungus, , by mutagenesis using ion beams or rays and evaluated the relative virulence of the resulting mutants compared with the wild-type isolates. In conclusion, ion beams and rays are useful tools for improving biological characteristics, such as thermotolerance and fungicide-tolerance, in entomopathogenic fungi, but the mutants obtained in this study must be carefully evaluated for unpredictable negative side effects.
荻野 千秋*; 山田 良祐*; 佐藤 勝也; 大野 豊
JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 122, 2015/03
Ethanol fermentation from hydrolyzed lignocellulosic biomass with yeast is very important. However, in common sense, there are several chemical inhibitors for yeast from hydrolyzed biomass. Therefore, screening of yeast strain, possessing the tolerance properties against these chemical inhibitors, should be found from nature resources or mutant libraries. In this research, by carbon ion beam irradiation, the screening of candidate yeast strains was performed. Compared with wild type strain, the ethanol production yield was improved in the case of selected strain. It was assumed that the carbon ion beam irradiation could be influence to yeast cell, and improve the metabolic pathway. In future, by using this selected yeast, the ethanol fermentation from hydrolyzed biomass with high efficiency would be performed.
増渕 隆*; 日向 弘和*; 上田 涼史郎; 林 秀謙*; 池永 裕*; 佐藤 勝也; 大野 豊
JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 123, 2015/03
We analyzed genome sequence of the high ethyl caproate producing sake yeast mutant (No.227) generated by ion beam breeding in order to investigated factors contributing the reduced ability to alcohol fermentation. In the high ethyl caproate producing sake yeast, four pyruvate decarboxylase genes (PDC), six alcohol dehydrogenase genes (ADH), two biotin synthesis genes (BIO) and chitinase gene (CTS1) might be involved in the reduced ability to alcohol fermentation. The genome sequence of the strain No.227 was determined by a whole-genome shotgun strategy using pyrosequencing method and compared with the whole-genome sequence of the sake yeast strain Kyokai 7, which is characterized by the fermentation property, as a reference sequence. For the PDC, ADH, BIO and CTS1 loci, no mutation was found in the strain No.227, suggesting that these genes did not involved in the reduced ability to alcohol fermentation.
布柴 達男*; 山内 理子*; 岩田 梨佳*; 佐藤 勝也; 大野 豊; 鳴海 一成*
JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 124, 2015/03
The LOH (loss of heterozygosity) induction by various ion beam radiations was investigated in diploid . The ion beams C, C, He and Ar at the lowest dose of 37.5 Gy, which had 10% lethality, induced LOH with 590%) resulted from homologous recombination. Pol may be involved in the induction of LOH by ion beam radiations, because in the strain lacking Rev3, only slight induction of LOH was observed, whereas no remarkable effect of deletion for Pol was observed on ion beam induced LOH. In addition, the effects of extracts were examined on iron-beam radiation induced LOH, and ethanol extract of possessed inhibitory effect on LOH induced by C, He and Arion-beam radiation.
Fitriana, Y.*; 篠原 忍*; 佐藤 勝也; 鳴海 一成*; 西東 力*
Applied Entomology and Zoology, 50(1), p.123 - 129, 2015/02
Fungicides targeted at phytopathogens can be harmful to entomopathogenic fungi. The purpose of this study was to use ion-beam irradiation technology to produce benomyl-resistant mutants of the entomopathogenic fungus . After irradiation of conidia at 150 Gy, two mutant isolates, BB22 and BB24, were selected on media containing the fungicide. In an assay of vegetative growth, BB22 and BB24 were over 500 and 800 times more tolerant to benomyl, respectively, compared with the wild isolate. A mutation was found at position 198 of the -tubulin gene in the mutant isolates, with a substitution of glutamate for alanine (E198A). Ion beams have great potential as a tool to improve the traits of entomopathogenic fungi such as increasing tolerance to fungicides. Fungicide-resistant mutants produced in this way could be useful agents for biological control within IPM programmes where fungicides are also used.
Attie, O.*; Jayaprakash, A.*; Shah, H.*; Paulsen, I. T.*; 守野 正人*; 高橋 優嘉*; 鳴海 一成*; Sachidanandam, R.*; 佐藤 勝也; 伊藤 政博*; et al.
Genome Announcements (Internet), 2(6), p.e01175-14_1 - e01175-14_2, 2014/11
The alkaliphilic bacterium AV1934, isolated from human feces, recent has indicated novel potassium ion coupling to motility in this extremophile. We report draft sequences that will facilitate an examination of whether that coupling is part of a larger cycle of potassium ion-coupled transporters. This draft genome has 4,095 predicted genes and 4,063 predicted proteins. The whole-genome shotgun project has been deposited in Genbank under the accession no. ALPT02000000. In addition to the genes encoding the MotPS channel that uses either potassium or sodium coupling, two loci encoding multisubunit Mrp-type antiporters, were found. The AV1934 genome reveals no tripartite ATP-independent (TRAP-T family) uptake systems,a major sodium-coupled complement in the other two alkaliphiles. The AV1934 genomic data will enable us to test the hypothesis of a major role of potassium in ion coupling in this extremophile.
安達 基泰; 平山 裕士; 清水 瑠美; 佐藤 勝也; 鳴海 一成*; 黒木 良太
Protein Science, 23(10), p.1349 - 1358, 2014/10
DNAの修復に関与する多機能なPprAは、デイノコッカスラジオデュランスの高度な放射線耐性を促進する重要なタンパク質である。PprAによる放射線耐性機構を解明するために、大腸菌で発現した組換え型PprAと2本鎖DNAとの相互作用解析を実施した。ゲルシフトアッセイにより、PprAとスーパーコイル型のpUC19 DNAとの複合体のゲルシフトが2極性であり、それが1mMのMg, Ca, Srイオンで促進されることが示された。シフトしたバンドの相対的な割合からPprAとスーパーコイル型のpUC19 DNAとの複合体形成のヒル係数および解離定数を計算したところ、1mM Mgイオンの存在下で、それぞれ3.3と0.6Mであった。このことは、PprAがスーパーコイル型のpUC19 DNAに少なくとも281分子結合していることを示しており、ゲル濾過での分離後にUV吸収で見積もられた値と一致した。この結果は、PprAが2本鎖DNAに沿って、直鎖状に重合して結合していることを示唆している。一方で、直鎖状の2本鎖DNAとニックがある環状DNAのバンドシフトに関しては、飽和が見られず、1.3M以上のPprA濃度の時、さらに大きな複合体の形成が確認された。この結果は、DNAに結合したPprAが、ダメージを受けたDNAの末端の会合を濃度依存的に促進していることを示している。
藤浪 俊*; 武田 喜代子*; 小野寺 威文*; 佐藤 勝也; 佐野 元彦*; 高橋 優嘉*; 鳴海 一成*; 伊藤 政博*
Genome Announcements (Internet), 2(5), p.e00866-14_1 - e00866-14_2, 2014/09
Calcium-dependent sp. strain TCA20 was isolated from a water sample of a hot spring containing a high concentration of calcium ions. Here, we report the draft genome sequence of this bacterium, which may be the basis for the research of calcium ion homeostasis. The draft genome sequence of sp. strain TCA20 totals 5,631,463 bp in length and is composed of 33 large contigs ( 500 bp) that was deposited at DDBJ/EMBL/Genbank under the accession no. BBIW00000000. The annotation of the draft genome sequence shows that sp. strain TCA20 has a calcium-specific calcium/proton antiporter, ChaA, and a P-type calcium-transporting ATPase, YloB, was identified as calcium transporters. These transporters may also be important for the growth of sp. strain TCA20 under the high concentration of calcium ions.