2020 Fiscal Year Final Research Report
Development of B12 analysis method for vitamins in foods using enzymatic quantification
| Project/Area Number |
18K14431
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| Research Category |
Grant-in-Aid for Early-Career Scientists
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| Allocation Type | Multi-year Fund |
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| Review Section |
Basic Section 38050:Food sciences-related
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| Research Institution | National Institutes of Biomedical Innovation, Health and Nutrition |
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Principal Investigator |
Suzuki Ippei 国立研究開発法人医薬基盤・健康・栄養研究所, 国立健康・栄養研究所 食品保健機能研究部, 研究員 (00812439)
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| Project Period (FY) |
2018-04-01 – 2021-03-31
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| Keywords | ビタミンB12 / リボヌクレオチドリダクターゼ / Lactobacillus / 微生物学的定量法 / 酵素学的定量法 / 乳酸菌 |
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| Outline of Final Research Achievements |
Research was conducted with the aim of developing a method for easily quantifying vitamin B12 (VB12) in foods by using enzymatic quantification. We investigated Ribonucleoside triphosphate reductase (RNR), which requires VB12 as a coenzyme. Recombinant RNR (reHis-RNR) with His-tag was purified. Using LC-MS, the production amount of dNTP, which is a reaction product, was used as an activity index. As a result, although it is necessary to further optimize the reHis-RNR reaction conditions in order to easily measure VB12 in foods, it is considered that there is some validity in using reHis-RNR activity as an index for measuring VB12 concentration.
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| Free Research Field |
食品分析
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| Academic Significance and Societal Importance of the Research Achievements |
乳酸菌由来ビタミンB12依存性リボヌクレオチドリダクターゼの大量発現系を構築するとともに、DNAシークエンス解析によって、変異個所を明らかにした。LC-MSを用いてリボヌクレオチドリダクターゼの反応産物であるdNTPの産生量を活性指標とした測定法を検討した結果、VB12を簡便に測定するためにはreHis-RNR反応条件のさらなる最適化が必要であるものの、より簡便なVB12測定法の開発に繋がると期待された。
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