Source: UNIVERSITY OF ILLINOIS submitted to
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
Funding Source
Reporting Frequency
Accession No.
Grant No.
Project No.
Proposal No.
Multistate No.
Program Code
Project Start Date
Feb 1, 2023
Project End Date
Jan 31, 2026
Grant Year
Project Director
Allen, J.
Recipient Organization
2001 S. Lincoln Ave.
Performing Department
Non Technical Summary
Fermented food intake is associated with improvements in human immune function. These benefits may lie in bioactivity of microbe-derived metabolites (i.e. postbiotics) found in fermented foods. It is our vision that 1) Identifying beneficial postbiotics and then 2) Adapting strategies to optimize their production in a food matrix will have significant impact on immune health and disease prevention. Our preliminary data indicates that select fermented foods contain high concentrations of phenyllactic acid (PLA), 4-hydroxyphenyllactic acid (4-HPLA), and indole-3-lactic acid (ILA); biochemically related metabolites downstream of microbial aromatic amino acid (ArAA) metabolism that we have identified as mediators of innate immune cell (monocyte) inflammatory activity. These data have led us to hypothesize that there is a unique class of microbial products within fermented food that confers anti-inflammatory activity. However, how fermented food postbiotic properties can be optimized to improve immune health of humans has never been tested. To fill this gap in knowledge, we will: 1) Further investigate the immune and metabolic tuning properties of microbial-ArAA metabolites 2) Identify food matrix conditions that optimize microbial ArAA metabolism and 3) Determine if whole fermented food diets optimized for high microbial-derived ArAA metabolites promote anti-inflammatory activity in humans.
Animal Health Component
Research Effort Categories

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
Knowledge Area
502 - New and Improved Food Products;

Subject Of Investigation
5010 - Food;

Field Of Science
1060 - Biology (whole systems);
Goals / Objectives
The primary goal of this projectis to determine if specific microbial-derived metabolites derived from aromatic amino acids (ArAALact) can be optimized in a whole food matrix to promote anti-inflammatory activity in humans. We will test this hypothesis with the following specific aims:Aim 1. Determine how microbial ArAALact metabolites directly mediate inflammatory and metatabolic profiles of primary human monocytes.Aim 2. Establish whole fermented food matrix conditions that maximize ArAALact metabolite production.Aim 3. Define effects of consuming whole fermented foods rich in ArAALact metabolites on circulating inflammatory profiles and the gut microbiota of obese humans.
Project Methods
For Aim 1 will we utilize ex vivo culture approaches with human monocytes to determine how microbial metabolites found in fermented food (ArAALact) promote anti-inflammatory activity. Briefly, monocytes will be isolated from healthy donors, cultured and stimulated with ArAALact. We will stimulate cells with bacterial components and then use pharmacological and siRNA-mediated blockades to test which pathways ArAALact may signal though. For Aim 2, we will utilize our expertise in food microbiology to determine how to optimize ArAALact in a whole food matrix. We will add various metabolic co-factors, precursor metabolites and substrates to bacterial cultures and fermented food matrices-followed by RSM based modeling to determine the optimal conditions for ArAALact production- both within individual bacterial cultures and whole foods. We will also expand on our data identifying commercial foods that contain the highest levels of ArAALact. Lastly, in Aim 3 we will define effects of consuming whole fermented foods rich in ArAALact on circulating inflammatory profiles in obese humans. This will be accomplished through a double-blinded fermented food feeding trial for eight weeksin obese humans. We will provide participants with either Diet 1) Lo-ArAALact -- Mixed fermented food meals with negligible levels of ArAALact metabolites or Diet 2) Hi-ArAALact --Mixed fermented food meals with high levels of ArAALact . Participants will receive onemix meal per day and twosnacks per day (112 total meals) across eightweeks. We will collect fasted blood and fecal samples before and after the eight-week period to test outcomes related to the gut microbiome and inflammatory status.

Progress 02/01/23 to 01/31/24

Target Audience:Our target audience for the USDA NIFA grant on optimizing bioactive metabolites in fermented foods includes both internal and external scientific communities. Internally, our research findings have been presented to scientific audiences at the University of Illinois, fostering collaboration and knowledge dissemination within the institution. Externally, our outreach efforts extended to other peerinstitutions, enhancing the impact and reach of our work. Furthermore, our commitment to knowledge dissemination is evident through the submission of a publication to BioRxiv, a preprint server, showcasing our research to a wider audience in the scientific community. This publication is nowcurrently under review at Nature-Science of Food. In summary, our target audience for this reporting periodcomprised mostlyscientists, researchers, and professionals in the field of Nutrition and related disciplines, both within our institution and beyond. Through our presentations and publications, we aim to share our progress and findings, fostering collaboration, and contributing to the advancement of knowledge of the potential health promoting properties offermented foods. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The project has significantly enriched the training and professional development opportunities for students in the Allen and Miller Lab, providing them with a multifaceted learning experience at the intersection of food science and immunology. Through active participation in the project, students were afforded the chance to receive hands-on training in cutting-edge techniques relevant to both fields, enhancing both their practical skills and theoretical knowledge. Notably, students were given the valuable opportunity to showcase their work and research findings to the broader nutrition science community through both oral and poster presentations. This exposure not only hones their communication and presentation skills but also fosters a deeper understanding of how their research contributes to the larger scientific landscape. Presenting at scientific gatherings, both internal and external, has empowered these students to engage with peers, experts, and professionals, establishing connections and gaining insights into diverse perspectives within nutritional sciences. Overall, the project has not only equipped students with technical expertise but has also instilled in them the ability to effectively communicate their findings, a crucial skill in their future careers. This dual emphasis on practical training and presentation opportunities reflects our commitment to nurturing the next generation of professionals in the fields of food science and immunology. How have the results been disseminated to communities of interest?Internally, within the University of Illinois, the project results have been shared through presentations at scientific forums and seminars. This includes both formal presentations and informal discussions, fostering dialogue and collaboration among researchers within the institution. Additionally, the presentation of multiple abstracts in poster format has provided a platform for interactive engagement and knowledge exchange within the academic community. Externally, our commitment to open science is evident through the publication of a pre-print paper in BioRxiv (December 2023), titled "Microbial aromatic amino acid metabolism is modifiable in fermented food matrices to promote bioactivity" by Kaseperk et al. This pre-print publication serves to disseminate our research findings to a global audience, allowing the broader scientific community to access and evaluate our work. Furthermore, the submission of a publication to Nature-Science of Food underscores our dedication to sharing our results with a prestigious journal, ensuring the dissemination of our findings to a wider and influential readership. Beyond academic circles, efforts have been made to communicate project outcomes to the general public and stakeholders through press releases, and summaries tailored for a non-specialist audience. This multi-faceted approach to dissemination aligns with our commitment to transparency, accessibility, and the widespread sharing of knowledge within communities of interest. What do you plan to do during the next reporting period to accomplish the goals?In the upcoming reporting period, we will channel our efforts towards the completion of Aim 1, focusing on definitively establishing the pathways through which ArAAlacts signal to monocytes. This will involve a comprehensive exploration and analysis to elucidate the intricate mechanisms underlying the interaction between ArAAlacts and monocytes, further enhancing our understanding of the biological processes at play. Simultaneously, as we conclude the remaining aspects of Aim 1, we will initiate the groundwork for Aim 3. The upcoming year will see the commencement of participant recruitment for our studies. Additionally, we will undertake the task of designing food menus tailored to the optimized fermented foods developed in the previous phases of the project. This menu design will lay the foundation for the upcoming cohort in the optimized fermented food feeding trial, scheduled to commence by the fall of 2024. The integration of Aim 1 completion and the initiation of Aim 3 represents a strategic progression in our research objectives. This dual focus ensures a comprehensive approach, combining the exploration of molecular pathways with the practical implementation of optimized fermented foods in human trials. Through these concerted efforts, we aim to deepen our understanding of the bioactive metabolites' impact on human health and inflammatory profiles, contributing valuable insights to the broader scientific community and advancing the goals of the USDA NIFA grant.

What was accomplished under these goals? In pursuit of Aim 1, significant strides have been made in establishing a robust model for culturing primary human monocytes ex vivo with ArAALact, accompanied by a well-defined workflow for subsequent gene expression and protein analysis. Additionally, a model for real-time monocyte metabolism analysis using Seahorse XF Analyzer has been successfully implemented. Although the pathways through antagonist treatments are yet to be defined, the groundwork has been laid for a comprehensive understanding. The outcomes of this research were disseminated through the presentation of multiple abstracts in poster format at both internal events within the University of Illinois and external scientific gatherings. In Aim 2, substantial progress has been achieved in establishing optimal conditions within the whole fermented food matrix to maximize ArAALact metabolite production. Our team meticulously analyzed concentrations of PLA, 4HPLA, ILA, and additional aromatic amino acid metabolites in over 25 commercially sold fermented foods, encompassing 15 dairy ferments, 10 vegetable ferments, and 4 miscellaneous ferments. A detailed examination of 12 fermented foods across two different lots was conducted. Furthermore, 21 lactic acid bacteria strains commonly found in fermented foods were assessed in monoculture, revealing Lactiplantibacillus plantarum as a robust producer of PLA, 4HPLA, and ILA. Techniques were developed to significantly enhance the production of these metabolites in two whole food matrices, yogurt and sauerkraut. The most optimized version of yogurt demonstrated remarkable increases, with over 2,000% in PLA, nearly 600% in 4HPLA, and over 3,000% in ILA compared to the yogurt control. A pre-print paper detailing these accomplishments was published in BioRxiv (December 2023), and a submission to Nature-Science of Food is currently under consideration. As we progress through the grant's first year, work on Aim 3 is pending initiation.The Institutional Review Board (IRB) approval has been successfully obtained. This aim focuses on defining the effects of consuming whole fermented foods rich in ArAALact metabolites on circulating inflammatory profiles and the gut microbiota of obese individuals, marking a critical avenue for future exploration.


  • Type: Journal Articles Status: Submitted Year Published: 2023 Citation: Kasperek, M. C. et al. Microbial aromatic amino acid metabolism is modifiable in fermented food matrices to promote bioactivity. bioRxiv, 2023.2012.2021.572869, doi:10.1101/2023.12.21.572869 (2023).