Sugar Shock: Probing Streptococcus pyogenes Metabolism Through Bioluminescence Imaging

• Published in: Frontiers in microbiology Vol. 13; p. 864014
• Main Authors: Davis, 4th, Richard W, Muse, Charlotte G, Eggleston, Heather, Hill, Micaila, Panizzi, Peter
• Format: Journal Article
• Language: English
• Published: Switzerland Frontiers Media S.A 02.06.2022
• Subjects: bioluminescence; biosensor; Microbiology; molecular imaging; pathogens; physiology; Streptococcus pyogenes; physiology; Streptococcus pyogenes; biosensor; pathogens; molecular imaging; bioluminescence
• ISSN: 1664-302X; 1664-302X
• DOI: 10.3389/fmicb.2022.864014

( ) can thrive in its host during an infection, and, as a result, it must be able to respond to external stimuli and available carbon sources. The preclinical use of engineered pathogens capable of constitutive light production may provide real-time information on microbial-specific metabolic processes. In this study, we mapped the central metabolism of a -modified Xen20 ( . Xen20) to its synthesis of luciferase substrates as assessed by the rate of light production in response to different environmental triggers. Previous characterization predicted that the operon was under the myo-inositol promotor. In this study, we revealed that supplementation with myo-inositol generated increased Xen20 luminescence. Surprisingly, when supplemented with infection-relevant carbon sources, such as glucose or glycine, light production was diminished. This was presumably due to the scavenging of pyruvate by -lactate dehydrogenase (LDH). Inhibition of LDH by its inhibitor, oxamate, partially restored luminescent signal in the presence of glucose, presumably by allowing the resulting pyruvate to proceed to acetyl-coenzyme A (CoA). This phenomenon appeared specific to the lactic acid bacterial metabolism as glucose or glycine did not reduce signal in an analogous -modified Gram-positive pathogen, . Xen29. The Xen20 cells produced light in a concentration-dependent manner, inversely related to the amount of glucose present. Taken together, our measures of microbial response could provide new information regarding the responsiveness of metabolism to acute changes in its local environments and cellular health.