Takayoshi Sekiguchi, Takako Sato, M. Enoki, H. Kanehiro, C. Kato
A procedure for the isolation of aliphatic polyester-degrading bacteria from deep-sea environments was developed using three step high-pressure cultivation. First, the sediments collected from the deep-sea floor, the Kurile and Japan Trenches at a depth of 5000-7000 m, were cultivated with poly e-caprolactone (PCL) films under low-temperature (4°C) and high-pressure (50MPa) conditions. Secondly, high-pressure continuous cultivation using the deep-sea baro-piezophile and thermophile isolation and cultivation (DEEPBATH) system was performed three times to accumulate piezophilic bacteria. Finally, piezophilic PCL degrading bacteria were isolated using the pressure bag method involving media with PCL granules. Using this procedure, we succeeded in isolating several piezophilic aliphatic polyester-degrading bacteria from the sediment samples of deep-trenches.
{"title":"Procedure for isolation of the plastic degrading piezophilic bacteria from deep-sea environments","authors":"Takayoshi Sekiguchi, Takako Sato, M. Enoki, H. Kanehiro, C. Kato","doi":"10.3118/JJSE.9.25","DOIUrl":"https://doi.org/10.3118/JJSE.9.25","url":null,"abstract":"A procedure for the isolation of aliphatic polyester-degrading bacteria from deep-sea environments was developed using three step high-pressure cultivation. First, the sediments collected from the deep-sea floor, the Kurile and Japan Trenches at a depth of 5000-7000 m, were cultivated with poly e-caprolactone (PCL) films under low-temperature (4°C) and high-pressure (50MPa) conditions. Secondly, high-pressure continuous cultivation using the deep-sea baro-piezophile and thermophile isolation and cultivation (DEEPBATH) system was performed three times to accumulate piezophilic bacteria. Finally, piezophilic PCL degrading bacteria were isolated using the pressure bag method involving media with PCL granules. Using this procedure, we succeeded in isolating several piezophilic aliphatic polyester-degrading bacteria from the sediment samples of deep-trenches.","PeriodicalId":204480,"journal":{"name":"Journal of Japanese Society for Extremophiles","volume":"99 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"115832879","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
H. Tamegai, T. Furukawa, J. Nitta, Sayaka Chikuma, M. Miyazaki, Y. Nogi, Shizuka Arakawa, C. Kato, K. Horikoshi
Moderately halophilic strain DT-W was isolated from the mud of the Mariana Trench. Cells of the organism were rod-shaped (1.5-2 m x 0.5-0.8 µm) with some flagella extruding from the cells. Growth occurred in an NaCl concentration of about 0.1-15% (optimal: 3-5%), at pH of 6-9, and at temperatures ranging from 4-51 °C (optimal: 30-37 °C). The results of 16S rDNA analysis and DNA-DNA hybridization analyses showed that DT-W was closely related to Halomonas aquamarina, Halomonas axialensis and Halomonas meridiana. Furthermore, additional physiological properties and the cytochrome contents of DT-W were also analyzed.
中度嗜盐菌株DT-W是从马里亚纳海沟的泥中分离出来的。细胞呈杆状(1.5-2m x 0.5-0.8µm)，细胞外伸出鞭毛。NaCl浓度为0.1-15%(最适为3-5%)，pH为6-9，温度为4-51°C(最适为30-37°C)。16S rDNA分析和DNA-DNA杂交分析结果表明，DT-W与海藻单胞菌、轴藻单胞菌和子午藻单胞菌亲缘关系密切。此外，还分析了DT-W的其他生理特性和细胞色素含量。
{"title":"Halomonas sp. strain DT-W, a halophile from the 11,000 m-depth of the Mariana Trench","authors":"H. Tamegai, T. Furukawa, J. Nitta, Sayaka Chikuma, M. Miyazaki, Y. Nogi, Shizuka Arakawa, C. Kato, K. Horikoshi","doi":"10.3118/JJSE.5.27","DOIUrl":"https://doi.org/10.3118/JJSE.5.27","url":null,"abstract":"Moderately halophilic strain DT-W was isolated from the mud of the Mariana Trench. Cells of the organism were rod-shaped (1.5-2 m x 0.5-0.8 µm) with some flagella extruding from the cells. Growth occurred in an NaCl concentration of about 0.1-15% (optimal: 3-5%), at pH of 6-9, and at temperatures ranging from 4-51 °C (optimal: 30-37 °C). The results of 16S rDNA analysis and DNA-DNA hybridization analyses showed that DT-W was closely related to Halomonas aquamarina, Halomonas axialensis and Halomonas meridiana. Furthermore, additional physiological properties and the cytochrome contents of DT-W were also analyzed.","PeriodicalId":204480,"journal":{"name":"Journal of Japanese Society for Extremophiles","volume":"4 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"131014490","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
To remediate the worldwide-spreading acid sulphate soil area and utilize it for human beings, acid tolerant leguminous plants are expected to be useful as crops and pioneer plants. The purpose of this study is to produce symbiotic nitrogen fixing systems tolerant to acid sulfate soils, through the identification of the genes that confer acid tolerance on plants and the isolation of acid-tolerant rhizobia. We identified acid tolerant genes by two approaches. One approach is functional screening of a soybean cDNA library and their characterization in planta. E. coli cells were transformed with a cDNA library of soybean and cultured on a LB medium adjusted to pH 4 or pH 5. Selected six tolerant clones were introduced into Arabidopsis thaliana using a plant expression vector. The overexpressors grew better than the wild type under the acid and aluminium stresses. The other one is molecular genetic analysis of the candidate genes identified by previous studies: a gene for a key enzyme of sulfur assimilation, cysteine synthase. The candidate gene was ectopically overexpressed in A. thaliana under the control of the cauliflower mosaic virus 35S promoter. All the genes tested were shown to confer acid tolerance on the model plants.
{"title":"Search for genes that confer acid tolerance to plants","authors":"T. Aoki, M. Shimamura, Y. Sawada, R. Imaizumi","doi":"10.3118/JJSE.7.2.20","DOIUrl":"https://doi.org/10.3118/JJSE.7.2.20","url":null,"abstract":"To remediate the worldwide-spreading acid sulphate soil area and utilize it for human beings, acid tolerant leguminous plants are expected to be useful as crops and pioneer plants. The purpose of this study is to produce symbiotic nitrogen fixing systems tolerant to acid sulfate soils, through the identification of the genes that confer acid tolerance on plants and the isolation of acid-tolerant rhizobia. We identified acid tolerant genes by two approaches. One approach is functional screening of a soybean cDNA library and their characterization in planta. E. coli cells were transformed with a cDNA library of soybean and cultured on a LB medium adjusted to pH 4 or pH 5. Selected six tolerant clones were introduced into Arabidopsis thaliana using a plant expression vector. The overexpressors grew better than the wild type under the acid and aluminium stresses. The other one is molecular genetic analysis of the candidate genes identified by previous studies: a gene for a key enzyme of sulfur assimilation, cysteine synthase. The candidate gene was ectopically overexpressed in A. thaliana under the control of the cauliflower mosaic virus 35S promoter. All the genes tested were shown to confer acid tolerance on the model plants.","PeriodicalId":204480,"journal":{"name":"Journal of Japanese Society for Extremophiles","volume":"3 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"131273655","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
M. Tanigawa, S. Seki, M. Mohri, A. Harigae, Y. Nagata
A NADPH dehydrogenase was purified from the hyperthermophilic archaeon, Pyrobaculum islandicum. The membrane protein was solubilized by treatment with 1% Tween 20, and purified 258-fold from the cell-free extract to homogeneity as judged by SDS-PAGE analysis. The molecular mass of the enzyme was revealed to be 41 kDa both by SDS-PAGE and gel filtration analyses. The optimal pH and temperature were 7.0 and above 80°C, respectively. The Km and Vmax values were 0.69 mM and 0.25 nmol/min, respectively. It was suggested that NADPH is an electron donor in the P. islandicum electron transfer system.
{"title":"Purification of NADPH dehydrogenase from a hyperthemophile, Pyrobaculum islandicum","authors":"M. Tanigawa, S. Seki, M. Mohri, A. Harigae, Y. Nagata","doi":"10.3118/JJSE.8.31","DOIUrl":"https://doi.org/10.3118/JJSE.8.31","url":null,"abstract":"A NADPH dehydrogenase was purified from the hyperthermophilic archaeon, Pyrobaculum islandicum. The membrane protein was solubilized by treatment with 1% Tween 20, and purified 258-fold from the cell-free extract to homogeneity as judged by SDS-PAGE analysis. The molecular mass of the enzyme was revealed to be 41 kDa both by SDS-PAGE and gel filtration analyses. The optimal pH and temperature were 7.0 and above 80°C, respectively. The Km and Vmax values were 0.69 mM and 0.25 nmol/min, respectively. It was suggested that NADPH is an electron donor in the P. islandicum electron transfer system.","PeriodicalId":204480,"journal":{"name":"Journal of Japanese Society for Extremophiles","volume":"61 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"117160288","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
75 - ， Abstract Having reached my retirement age ， I retired from Kagawa Prefectural University of Health Sciences on March 31, 2014 ． The main area of my research interest for 40 years was endocrinology ． In this article ， I look back on my encounters with people who kindly supported my research ． In addition ， I summarize the four main themes of my recent research ：( 1 ） Central mechanisms of appetite regulation ；( 2 ） Clinical research related to obesity ；( 3 ） Role of neuropeptides related to stress ；( 4 ） Methods to evaluate stress and their clinical application ． I sincerely thank Kagawa Prefectural University of Health Sciences for giving me the opportunity to contribute to this journal.
{"title":"Looking back on my research life.","authors":"R. Iizuka","doi":"10.3118/JJSE.8.15","DOIUrl":"https://doi.org/10.3118/JJSE.8.15","url":null,"abstract":"75 - ， Abstract Having reached my retirement age ， I retired from Kagawa Prefectural University of Health Sciences on March 31, 2014 ． The main area of my research interest for 40 years was endocrinology ． In this article ， I look back on my encounters with people who kindly supported my research ． In addition ， I summarize the four main themes of my recent research ：( 1 ） Central mechanisms of appetite regulation ；( 2 ） Clinical research related to obesity ；( 3 ） Role of neuropeptides related to stress ；( 4 ） Methods to evaluate stress and their clinical application ． I sincerely thank Kagawa Prefectural University of Health Sciences for giving me the opportunity to contribute to this journal.","PeriodicalId":204480,"journal":{"name":"Journal of Japanese Society for Extremophiles","volume":"28 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"133963746","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Regulation and environmental response of aromatic degradation gene expression in a PCB degrading bacterium.","authors":"M. Fukuda","doi":"10.3118/JJSE.6.63","DOIUrl":"https://doi.org/10.3118/JJSE.6.63","url":null,"abstract":"","PeriodicalId":204480,"journal":{"name":"Journal of Japanese Society for Extremophiles","volume":"1 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"132526216","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
T. Matsuura, T. Terayama, R. Usami, H. Yoshida, H. Hara, K. Matsumoto
To understand the physiological role of the ykoN gene of Bacillus subtilis, the gene was expressed from the IPTG-inducible spac promoter in B. subtilis cells. When lipid composition of the cells induced for ykoN expression was examined, a new spot of phospholipid was found on the thin layer chromatogram. Induction of ykoN in Escherichia coli cells produced a similar new spot. The new spot was produced in mutant E. coli cells lacking cardiolipin or phosphatidylethanolamine, but not in the cells lacking phosphatidylglycerol. The result suggests an involvement of phosphatidylglycerol in production of the new spot. Examination of the lipid composition during the stages of B. subtilis growth revealed that the new phospholipid was produced four hours after cessation of logarithmic growth, consistent with an increase in the promoter activity of ykoN in the late stage in sporulation. YkoN has the pentapeptide lipase/esterase motif (Gly-X-Ser-X-Gly), and shows a lipolytic activity (Matsuura et al., in press). The mutant YkoN (S205A) in which the central serine residue of the motif is replaced with alanine produced the new phospholipid in the amount as small as only 1/10 of the wild type, suggesting that the lipolytic activity of YkoN is involved in the new phospholipid production.
{"title":"Involvement of YkoN in production of a new phospholipid in Bacillus subtilis Marburg","authors":"T. Matsuura, T. Terayama, R. Usami, H. Yoshida, H. Hara, K. Matsumoto","doi":"10.3118/JJSE.6.38","DOIUrl":"https://doi.org/10.3118/JJSE.6.38","url":null,"abstract":"To understand the physiological role of the ykoN gene of Bacillus subtilis, the gene was expressed from the IPTG-inducible spac promoter in B. subtilis cells. When lipid composition of the cells induced for ykoN expression was examined, a new spot of phospholipid was found on the thin layer chromatogram. Induction of ykoN in Escherichia coli cells produced a similar new spot. The new spot was produced in mutant E. coli cells lacking cardiolipin or phosphatidylethanolamine, but not in the cells lacking phosphatidylglycerol. The result suggests an involvement of phosphatidylglycerol in production of the new spot. Examination of the lipid composition during the stages of B. subtilis growth revealed that the new phospholipid was produced four hours after cessation of logarithmic growth, consistent with an increase in the promoter activity of ykoN in the late stage in sporulation. YkoN has the pentapeptide lipase/esterase motif (Gly-X-Ser-X-Gly), and shows a lipolytic activity (Matsuura et al., in press). The mutant YkoN (S205A) in which the central serine residue of the motif is replaced with alanine produced the new phospholipid in the amount as small as only 1/10 of the wild type, suggesting that the lipolytic activity of YkoN is involved in the new phospholipid production.","PeriodicalId":204480,"journal":{"name":"Journal of Japanese Society for Extremophiles","volume":"31 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"115698368","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Tadamasa Fukushima, A. Echigo, K. Fukuda, R. Usami
More than 80% of residual halophilic α-amylase (crude and purified) activity from strain S-1 was observed in the presence of various organic solvents. More than 78% remaining activity was detected after incubation for 120 hr in the presence of styrene, toluene, benzene or chloroform. Enzyme activity and stability with chloroform were higher than that without chloroform at the low or high temperature, and enzyme activity was higher in the presence of chloroform than one without chloroform when EDTA (5-15 mM) was added.
{"title":"Effect of organic solvents, temperature and EDTA on activity and stability of halophilic .ALPHA.-amylase from Haloarcula sp. S-1","authors":"Tadamasa Fukushima, A. Echigo, K. Fukuda, R. Usami","doi":"10.3118/JJSE.4.56","DOIUrl":"https://doi.org/10.3118/JJSE.4.56","url":null,"abstract":"More than 80% of residual halophilic α-amylase (crude and purified) activity from strain S-1 was observed in the presence of various organic solvents. More than 78% remaining activity was detected after incubation for 120 hr in the presence of styrene, toluene, benzene or chloroform. Enzyme activity and stability with chloroform were higher than that without chloroform at the low or high temperature, and enzyme activity was higher in the presence of chloroform than one without chloroform when EDTA (5-15 mM) was added.","PeriodicalId":204480,"journal":{"name":"Journal of Japanese Society for Extremophiles","volume":"76 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"123827480","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Study on nitrogen cycling by using stable isotope techniques","authors":"K. Koba","doi":"10.3118/JJSE.6.52","DOIUrl":"https://doi.org/10.3118/JJSE.6.52","url":null,"abstract":"","PeriodicalId":204480,"journal":{"name":"Journal of Japanese Society for Extremophiles","volume":"21 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"122230062","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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