Isotopic tracer for absolute quantification of metabolites of the pentose phosphate pathway in bacteria

Document Type

Article

Publication Date

11-1-2022

Abstract

The pentose phosphate pathway (PPP) plays a key role in many metabolic functions, including the generation of NADPH, biosynthesis of nucleotides, and carbon homeostasis. In particular, the intermediates of PPP have been found to be significantly perturbed in bacterial metabolomic studies. Nonetheless, detailed analysis to gain mechanistic information of PPP metabolism remains limited as most studies are unable to report on the absolute levels of the metabolites. Absolute quantification of metabolites is a prerequisite to study the details of fluxes and its regulations. Isotope tracer or labeling studies are conducted in vivo and in vitro and have significantly improved the analysis and understanding of PPP. Due to the laborious procedure and limitations in the in vivo method, an in vitro approach known as Group Specific Internal Standard Technology (GSIST) has been successfully developed to measure the absolute levels of central carbon metabolism, including PPP. The technique adopts derivatization of an experimental sample and a corresponding internal standard with isotope-coded reagents to provide better precision for accurate identification and absolute quantification. In this review, we highlight bacterial studies that employed isotopic tracers as the tagging agents used for the absolute quantification analysis of PPP metabolites.

Keywords

Metabolomic, Bacterial metabolomics, Pentose phosphate pathway (PPP), Isotope labeling, Absolute quantification

Divisions

clinicalpharmacy

Funders

Ministry of Education, Malaysia under the Fundamental Research Grant Scheme (FRGS),International Islamic University Malaysia (IIUM) (Grant No: FRGS/1/2019/STG03/UIAM/03/1 & FRGS19-119-0728)

Publication Title

Metabolities

Volume

12

Issue

11

Publisher

MDPI

Publisher Location

ST ALBAN-ANLAGE 66, CH-4052 BASEL, SWITZERLAND

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