To understand microbial metabolic functionality, it is important to focus on intracellular reaction kinetics. This requires the development of methodologies to measure intracellular reactant concentrations under in vivo conditions. The analysis of intracellular metabolites, nucleotides, and cofactors has been attempted with a wide range of techniques, e.g., enzymatic assays, HPLC, GC-MS, LC-MS, and NMR. Although there are some limitations in using ESI LC-MS detection, it has a number of advantages compared to the other methods. Since glycolytic intermediates are especially difficult to analyze due to their polarity, structural similarity, and noncharacteristic UV absorption. MS detection has not only the advantage of specific detection, but also that has m/z-specific separation. Here we propose a convenient and highly selective and sensitive LCMS/MS method using tributylamine as volatile ion pair reagent. The method allows the combined separation and quantification of 9 metabolites from the most interesting biological classes in central carbon metabolism. This method can be successfully applied to identification and quantification of these intracellular metabolites in various cell extract samples. In this study determination of intracellular metabolites in E. coli was given as an example.
한국생물공학회 [The Korean Society for Biotechnology and Bioengineering]
설립연도
1984
분야
공학>생물공학
소개
이 법인은 생물 공학의 발전과 보급에 이바지하고, 회원 상호 간의 연구 협력과 친목을 도모함을 목적으로 한다
1. 생물공학 분야의 발전을 위한 연구 협력
2. 생물공학의 실용화를 촉진시키기 위한 산학 협동
3. 학술연구 발표회, 강연회, 연수회 등 학술활동의 개최
4. 국,영문 학술지,소식지,학술회의 Proceedings 및 학술도서의 발간
5. 생물공학 발전을 위한 정책 건의
6. 기타 국제 교류 등 본 학회의 목적 달성을 위한 제반 활동