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Characterizing fatty acid uptake, metabolism, and storage by gut bacteria from the genus Turicibacter

Award Number: F31AI200223
ORGANIZATION: NATIONAL INSTITUTE OF ALLERGY & INFECTIOUS DISEASES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: FELLOWSHIP/SCHOLARSHIP/STUDENT LOANS
PERIOD OF PERFORMANCE START DATE: 06/15/2026
PERIOD OF PERFORMANCE END DATE: 06/14/2029

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Characterizing fatty acid uptake, metabolism, and storage by gut bacteria from the genus Turicibacter - PROJECT SUMMARY/ABSTRACT The gastrointestinal microbiota is a potent mediator of host lipid interactions and metabolism. Gram positive bacteria from the understudied Turicibacter genus live in the guts of mammals, birds, and fish and are predicted to interact with lipids in human and animal hosts. Monocolonization of mice with different strains of Turicibacter alter host serum lipids with a notable decrease in serum triglycerides, particularly those containing saturated and monounsaturated fatty acids (FAs). To understand how Turicibacters directly interact with FAs, I performed RNAseq on Turicibacter cultures preincubated with FAs and found that saturated, monounsaturated, and polyunsaturated FAs elicit distinct transcriptional responses in these cells. These results led me to identify putative Turicibacter gene homologs of a recently discovered Gram positive FA uptake system called the fatty acid kinase (Fak) system. My RNAseq data indicated that different FA substrates prompted a different regulation pattern for the four FA-binding (FakB) homologs possessed by Turicibacters, and later amino acid sequence analysis revealed that three of these homologs are distinct from all previously studied Fak proteins. Aim 1 of my proposal is to define the mechanism of FA uptake by Turicibacters and characterize FA binding and specificity of novel FakB proteins. To do this, I will purify Fak kinase and FakB homologs identified in Turicibacters, then probe their ability to phosphorylate FAs (a crucial first step during metabolism) and conduct FA binding affinity and competition assays to determine their binding specificity. This characterization of the Turicibacter Fak system will improve our understanding of FA uptake across Gram positive bacteria. Aim 2 of my proposal will define the consequences of Turicibacter FA uptake by studying lipidomic and metabolomic changes in Turicibacter cells and subcellular lipid droplet (LD)-like structures during growth with FAs. Initial experiments using fluorescent labeled FAs indicate that Turicibacters incorporate exogenous FAs into polar and neutral lipids, and imaging analysis revealed the presence of lipid-labeled structures within Turicibacter cells that are not well studied in gut bacteria. I will first study changes in whole cells when Turicibacters are incubated with different FAs, then isolate LD-like structures and identify their specific lipid and protein components. Finally, I will use fluorescent labeling and immunofluorescence to study the co-occurrence of Fak proteins with lipid accumulation and cell growth. These experiments will allow us to identify specific products of FAs taken up by Turicibacters and provide clues to the function of lipid storage structures in environments where nutrients are abundant. Together, these aims will characterize how Turicibacters take up, metabolize, and store exogenous FAs. The results of these studies will lay a foundation for future research into how FA metabolism by Turicibacters and related bacteria influences host lipid metabolism and inform production of microbiota-based therapeutics for metabolic disorders.


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2026 ( Subtotal = $50,114 )
2026	2026	THE JOHNS HOPKINS UNIVERSITY	3400 N CHARLES ST	BALTIMORE	MD	21218	BALTIMORE CITY	USA	Allergy and Infectious Diseases Research	000	1	5/7/2026	NEW	$50,114
2026	2026	THE JOHNS HOPKINS UNIVERSITY	3400 N CHARLES ST	BALTIMORE	MD	21218	BALTIMORE CITY	USA	Allergy and Infectious Diseases Research	001	1	5/7/2026	NEW	$0
														Subtotal = $50,114

Grand Total All Awards = $50,114

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Characterizing fatty acid uptake, metabolism, and storage by gut bacteria from the genus Turicibacter

Award Number: F31AI200223

ORGANIZATION: NATIONAL INSTITUTE OF ALLERGY & INFECTIOUS DISEASES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: FELLOWSHIP/SCHOLARSHIP/STUDENT LOANS

PERIOD OF PERFORMANCE START DATE: 06/15/2026

PERIOD OF PERFORMANCE END DATE: 06/14/2029

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