GC/MS을 이용한 Yersinia enterocolitica 균체 및 배지의 대사체 분석
GC/MS-based metabolomic analysis of Yersinia enterocolitica and its growth media
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초록
배양시간(6, 12, 24, 48시간)에 따른 Yersinia enterocolitica 균체 및 배지 대사체들의 변화를 이해하기 위하여 GC/MS와 다변량통계분석을 이용하여 분석한 결과 Partial Least Squares-DiscriminantAnalysis(PLS-DA) scores plots 상에서 배양시간에 따라 분석 시료들이 뚜렷하게 분리되었다. 시료들사이의 차이에 관여하는 대사물질들을 동정한 결과 균체에서는 ethylene glycol, valine,ethanolamine, succinic acid, adenine, stearic acid, 배지에서는 glycine, fumaric acid, threonine,aminomalonic acid, malic acid, glutamic acid, citric acid가 배양시간에 따른 차이가 나는 것으로확인되었다. 이들 대사물질들을 이용하여 관련 대사경로를 도출한 결과 배양시기에 따라 에너지 생성에관여하는 대사경로가 주로 변하는 것으로 관찰되었다. Y. enterocolitica의 경우 대수기 초기에는 TCAcycle을 통해 에너지를 공급하다가 정지기에 들어서면서 정상적인 TCA cycle를 수행하지 못하고ethanolamine 및 ethylene glycol 등 다른 탄소원 또는 질소원을 공급하는 것으로 확인되었다. 비록관련된 많은 연구가 필요함에도 불구하고 대사체분석 기술을 활용한 Y. enterocolitica 균체 및 배지분석 연구는 배양시간에 따른 대사과정의 변화를 관찰할 수 있을 뿐만 아니라 향후 균체 연구의 다양한분야에 접목될 수 있을 것으로 사료된다.
Abstract
To better understand microbial metabolism, metabolomic profiles of Yersinia enterocolitica andits growth media at the different growth stages(6, 12, 24, and 48h) were analyzed using gaschromatography-mass spectrometry(GC/MS). The collected data were statistically analyzed andvisualized by Partial Least Squares-Discriminant Analysis(PLS-DA). Significant differences amongsample groups were observed on the PLS-DA scores plot. The differences of thirteenmetabolites including amino acids, organic acids related to the TCA cycle, ethylene glycol, andethanolamine were observed over the growth stage. Based on these metabolites, the metabolic pathway associated with the growth stage of Y. enterocolitica has been proposed. Energy sourcesof Y. enterocolitica might be changed according to the growth stage. This metabolomic studysuggested that energy at the log phase might be produced via the TCA cycle but after thestationary phase, and thus the TCA cycle was dysregulated and other carbon and/or nitrogensources such as ethylene glycol and etanolamine might be required.
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