Stable Isotope-Assisted Metabolic Profiling Reveals Growth Mode Dependent Differential Metabolism and Multiple Catabolic Pathways of l-Phenylalanine in Rubrivivax benzoatilyticus JA2,Journal of Proteome Research

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Stable Isotope-Assisted Metabolic Profiling Reveals Growth Mode Dependent Differential Metabolism and Multiple Catabolic Pathways of l-Phenylalanine in Rubrivivax benzoatilyticus JA2
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2017-11-07 00:00:00 , DOI: 10.1021/acs.jproteome.7b00500
Lakshmi Prasuna Mekala ₁ , Mujahid Mohammed ₁ , Sasikala Chintalapati ₂ , Venkata Ramana Chintalapati ₁

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Anoxygenic phototrophic bacteria are metabolically versatile and survive under different growth modes using diverse organic compounds, yet their metabolic diversity is largely unexplored. In the present study, we employed stable-isotope-assisted metabolic profiling to unravel the l-phenylalanine catabolism in Rubrivivax benzoatilyticus JA2 under varying growth modes. Strain JA2 grows under anaerobic and aerobic conditions by utilizing l-phenylalanine as a nitrogen source. Furthermore, ring-labeled ¹³C₆-phenylalanine feeding followed by liquid chromatography–mass spectrometry exometabolite profiling revealed 60 labeled metabolic features (M + 6, M + 12, and M + 18) derived solely from l-phenylalanine, of which 11 were identified, 7 putatively identified, and 42 unidentified under anaerobic and aerobic conditions. However, labeled metabolites were significantly higher in aerobic compared to anaerobic conditions. Furthermore, detected metabolites and enzyme activities indicated multiple l-phenylalanine catabolic routes mainly Ehrlich, homogentisate-dependent melanin, benzenoid, and unidentified pathways operating under anaerobic and aerobic conditions in strain JA2. Interestingly, the study indicated l-phenylalanine-dependent and independent benzenoid biosynthesis in strain JA2 and a differential flux of l-phenylalanine to Ehrlich and benzenoid pathways under anaerobic and aerobic conditions. Additionally, unidentified labeled metabolites strongly suggest the presence of unknown phenylalanine catabolic routes in strain JA2. Overall, the study uncovered the l-phenylalanine catabolic diversity in strain JA2 and demonstrated the potential of stable isotope-assisted metabolomics in unraveling the hidden metabolic repertoire.

中文翻译：

稳定同位素辅助代谢谱揭示增长模式相关的差异代谢和L-苯丙氨酸的多个代谢途径Rubrivivax benzoatilyticus JA2

产氧的光养细菌具有多种代谢功能，可以使用多种有机化合物在不同的生长模式下生存，但其代谢多样性尚未得到充分开发。在本研究中，我们采用了稳定同位素辅助的代谢谱分析方法，揭示了在不同生长模式下Rubrivivax benzoatilyticus JA2中的l-苯丙氨酸分解代谢。通过利用1-苯丙氨酸作为氮源，菌株JA2在厌氧和有氧条件下生长。此外，通过环标记的¹³ C _6-苯丙氨酸进料，然后进行液相色谱-质谱联生代谢产物谱分析，发现60种标记的代谢特征（M + 6，M + 12和M + 18）仅来源于l-苯丙氨酸，在厌氧和有氧条件下被鉴定出11个，推定被鉴定出7个，未鉴定出42个。但是，与无氧条件相比，有氧代谢中标记的代谢物明显更高。此外，检测到的代谢物和酶活性表明，JA2菌株在厌氧和好氧条件下有多种1-苯丙氨酸分解代谢途径，主要是Ehrlich，依赖尿黑酸的黑色素，Benzenoid和未知途径。有趣的是，该研究表明菌株JA2中l-苯丙氨酸依赖性和非依赖性的苯甲酰生物合成以及l的差异通量。厌氧和好氧条件下，β-苯丙氨酸到Ehrlich和Benzenoid的途径。另外，未鉴定的标记代谢物强烈提示菌株JA2中存在未知的苯丙氨酸分解代谢途径。总体而言，该研究揭示了菌株JA2中的l-苯丙氨酸分解代谢多样性，并证明了稳定的同位素辅助代谢组学在揭示隐藏的代谢库方面的潜力。

更新日期：2017-11-07

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