2014 Fiscal Year Annual Research Report
| Project/Area Number |
25292156
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| Research Institution | Kobe University |
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Principal Investigator |
ルミアナ ツエンコヴア 神戸大学, (連合)農学研究科(研究院), 教授 (30294200)
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| Co-Investigator(Kenkyū-buntansha) |
近江戸 伸子 神戸大学, 人間発達環境学研究科, 教授 (30343263)
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| Project Period (FY) |
2013-04-01 – 2016-03-31
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| Keywords | aquaphotomics / NIRS / cells / probiotics / water |
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| Outline of Annual Research Achievements |
This year, beside some fundamental experiments such as evaluation of the water structure in low concentration solutes including aqueous solution of sugar, salt, acid, DNA and mineral water samples, our focus has been on the analyses of Lactobacillus bacteria strains.
VIS-NIR spectra of 25 different Lactobacillus strains were acquired at, both, dynamic and static conditions. Monitoring of the Lactobacillus strains during their 24 hours cultivation time in MRS broth at 37 oC was performed to observe the water structural changes in their natural environment. Experiments were also conducted to determine the water structure evolved by the bacteria cells and their metabolites in the stationer phase of the growing curve.
Our results showed that the optimal cultivation time for the evaluation of the differences between the strains having different functionalities is in the exponential growth phase. Sophisticated chemometrics methods including OPLS-DA, SIMCA, PLSR provided highly accurate models for classification (100%) of probiotic and non-probiotic Lactobacillus strains and to predict their main characteristics such as MIC of Bile and Yield of biomass after 3 h stay at pH 1.80 and 9000 U/ml pepsin (R square: 0.79, 0,84 and RMSEP: 0.243mg/ml, 0.018, respectively). The results also present the importance of water bands in the developed models. The assigned bands showed that the differences among the strains having different probiotic strength are mainly due to the presence of free water molecules, water solvation shells and protonated water.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
The last two years of the project we developed spectral data base for somatic and callus rice (first year) and (probiotic, moderate and non-probiotic) Lactobacillus bacterial strains (the second year)that consist of spectra acquired in the process of cell development. Applying multivariate spectral data analysis, we were able to find specific common water bands, WAMACS, for the cells and to identify non-invasively the developmental stages for all types of cells based on newly identified common WAMACS. It was possible to discriminate probiotic, moderate and non-probiotic Lactobacillus bacterial cells by analyzing their spectral data acquired in the process of cell development and find their specific spectral patterns. We found that spectra acquired at the 10th hour of cell development are the best spectral data for Lacto Bacillus bacteria identification. We have found the most specific water molecular system conformations related to bacteria strain identification, i.e. to the functionality of bacterial ssystem. The group of probiotic bacteria characterises with higher number of small protonated water clusters, more free water molecules and water clusters with weak hydrogen bonds when compared with the other two groups. The group of probiotic bacteria also show big absorption in the region of water-protein interactions followed by the moderate and non-probiotic strains. In the last year, we have submitted one book chapter, 8 papers and 14 presentations at national and international conferences.
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| Strategy for Future Research Activity |
In the last year of the project we will concentrate on expanding the existing data base with new experiments and spectral data acquisition of spectra of somatic and callus rice and bacteria adding new bacteria types. Having all these results will allow us to build up general cell’s WAMACS data base and to find connections between specific spectral patterns and developmental stages, species and strains difference. We will compare the obtained data with the spectral data base that we have previously acquired (2011 kaken project on DNA and single protein aqueous solutions) in order to understand the commonality between water specific bands and spectral patterns of simple aqueous systems and cell systems. Our final goal is to relate water spectral patterns and even single water bands to biological functions.
We are planing to collaborate woth other laboratories in Japan and in the world in order to repeat our experiments and investigate similar systems. Different climate and environment conditions miight influence differently the water matrix. Therefore, we need to investigate if we can still find the same WAMCS and water spectral patterns related to the same functions, strain characteristics.
Another direction is to work on the development of new sensors that can measure directly water spectral patterns of the investigated system and directly relate it to its function or specific characteristics.It will allow automation and non-destructive measurement of cell viability measurement and cell identification.
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| Causes of Carryover |
購入物品が予定より安価にて購入できたため
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| Expenditure Plan for Carryover Budget |
物品購入
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[Presentation] Research of early flowering transgenic Jatropha plant2015
Author(s)
Ayaka Hata,Suguru Tsuchimoto, Kiichi Fukui, Hisashi Tomemori, Hisashi Tsujimoto, Nobuko Ohmido
Organizer
8th International Symposium Exploring the global sustainability-Advances in Plant Biotechnology for Agriculture in Semi-ariland
Place of Presentation
大阪大学
Year and Date
2015-03-03 – 2015-03-04
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