- 著者
- Akihiko Yamagishi Shin-ichi Yokobori Yoshitaka Yoshimura Masamichi Yamashita Hirofumi Hashimoto Takashi Kubota Hajime Yano Junichi Haruyama Makoto Tabata Kensei Kobayashi Hajime Honda Yuichi Utsumi Tsunemasa Saiki Takashi Itoh Atsuo Miyakawa Kenji Hamase Takeshi Naganuma Hajime Mita Kenichi Tonokura Sho Sasaki Hideaki Miyamoto
- 出版者
- 日本宇宙生物科学会
- 雑誌
- Biological Sciences in Space (ISSN:09149201)
- 巻号頁・発行日
- vol.24, no.2, pp.67-82, 2010 (Released:2012-06-26)
- 参考文献数
- 114
- 被引用文献数
- 1 12
Liquid water is considered to be critical for life. Gibbs free energy is another factor that is important to sustain life for long durations. Gibbs free energy is obtained by reactions between reductants and oxidants, or from any other non-equilibrium state of matter. As an example, aerobic organisms use carbohydrates and oxygen to obtain energy. Many types of chemoautotrophic mechanisms are known for this process as well. On the surface of Mars, methane and oxidative compounds such as ferric oxide, sulfate and perchloride, which could provide redox-derived Gibbs free energy, have been detected. Iron-dependent methane oxidizing bacteria have been found in marine environments on Earth. This finding suggests the possible presence of methane-oxidizing bacteria on the Mars surface, if the local thermal environment and other resources permit proliferation and metabolism of bacteria. Our project aims to search for methane-oxidizing microbes on the Mars surface. Martian soil will be sampled from a depth of about 5 - 10 cm below the surface, where organisms are expected to be protected from the harsh hyper-oxidative environment of the Mars surface. Small particles less than 0.1 mm or 1 mm will be sieved from the sample, before being transferred to the analysis section by a micro-actuator. The particles will be stained with a cocktail of fluorescent reagents, and examined by fluorescence microscopy. A combination of fluorescent dyes has been selected to identify life forms in samples. A membrane-specific dye or a combination of dyes will be used to detect membranes surrounding the "cell". An intercalating fluorescent dye such as SYBR Green will be used to detect genetic compounds such as DNA. A substrate dye that emits fluorescence upon cleavage by a catalytic reaction will be used to detect the catalytic activity of the "cell". A combination of staining reagents has been chosen based on the definition of life. A membrane separating a cell from the ambient environment may lead to identification of an "individual". DNA or genetic material is required for "replication" of the life form. Catalytic reactions carried out by enzymes drive "metabolism". This combination of strategies will also be useful for detecting pre-biotic organic material as well as remnants of ancient life. Hydrolysis of the polymers in the "cell" followed by HPLC or soft ionization MS for amino acid analysis will be effective for examining whether Martian life is identical to or different from terrestrial life. The number and type of the amino acids as well as their chirality will be analyzed to distinguish whether the polymers are contaminants from Earth.
- 著者
- Chiharu ISHII Tetsuya MIYAMOTO Shoto ISHIGO Yurika MIYOSHI Masashi MITA Hiroshi HOMMA Tadashi UEDA Kenji HAMASE
- 出版者
- クロマトグラフィー科学会
- 雑誌
- CHROMATOGRAPHY (ISSN:13428284)
- 巻号頁・発行日
- pp.2017.009, (Released:2017-06-04)
- 参考文献数
- 46
- 被引用文献数
- 15
The formation of D-amino acid residues in proteins is considered as one of the deterioration processes, and the determination of these D-amino acid residues is highly expected for the screening of new biomarkers under various disease conditions. In the present study, a two-dimensional (2D) HPLC-MS/MS system following the hydrolysis with deuterium chloride (2HCl/2H2O) and derivatization of amino acids with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) has been designed/developed and applied to the analysis of proteins stored under various conditions. As the target, 5 major D-amino acid residues (Ala, Asp, Glu, Pro and Ser) were selected. The analytical procedure was validated using a model peptide, NH2-Gly-Pro-Glu-Ala-Asp-Ser-Gly-OH, and the obtained calibration lines of %D for the 5 target amino acids were linear with correlation coefficients greater than 0.998. The RSD values for the intra-day precision and inter-day precision were lower than 5%. In most of the proteins tested, the amounts of the D-Ser and D-Asp residues increased during storage, and the highest value (14%, D-Ser) was observed in ovalbumin (OVA) after storage at pH 9.5 for 4 weeks.
- 著者
- Kenji HAMASE Yusuke NAKAUCHI Yurika MIYOSHI Reiko KOGA Nao KUSANO Hirohisa ONIGAHARA Hiroshi NARAOKA Hajime MITA Yasuhiko KADOTA Yasuhiro NISHIO Masashi MITA Wolfgang LINDNER
- 出版者
- クロマトグラフィー科学会
- 雑誌
- CHROMATOGRAPHY (ISSN:13428284)
- 巻号頁・発行日
- vol.35, no.2, pp.103-110, 2014-08-10 (Released:2014-08-26)
- 参考文献数
- 39
- 被引用文献数
- 4 33
A two-dimensional chiral high-performance liquid chromatographic (2D-HPLC) system has been established for the analysis of extraterrestrial amino acids. As the targets, 8 chiral amino acids (alanine (Ala), valine (Val), 2-aminobutyric acid (2AB), norvaline (nVal), N-methylalanine (N-MeAla), isovaline (iVal), 3AB and 3-aminoisobutyric acid (3AIB)) and 5 non-chiral amino acids (glycine (Gly), β-Ala, γ-aminobutyric acid (GABA), sarcosine (Sar) and 2AIB) were selected. These amino acids were tagged with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), and non-enantioselectively separated by a capillary monolithic ODS column in the first dimension. The target fractions were automatically introduced into the second dimension and further separated by Pirkle-type enantioselective columns. By using this system, the 2D-HPLC separation of 21 components in small particles of a carbonaceous chondrite (Yamato 791191, Antarctic CM2 meteorite) could be successfully performed, and all of the target amino acids were observed. The D/L ratios of the chiral molecules are almost 50/50 for all of the tested proteinogenic and non-proteinogenic amino acids.