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Development of B12 analysis method for vitamins in foods using enzymatic quantification

Research Project

Project/Area Number 18K14431
Research Category

Grant-in-Aid for Early-Career Scientists

Allocation TypeMulti-year Fund
Review Section Basic Section 38050:Food sciences-related
Research InstitutionNational Institutes of Biomedical Innovation, Health and Nutrition

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)

Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2019: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2018: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
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 / 酵素精製 / RNR / ビタミン12 / VB12 / 分析法の開発
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.

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。

Report

(4 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Research-status Report
  • 2018 Research-status Report

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Published: 2018-04-23   Modified: 2022-01-27  

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