Progress 06/01/23 to 05/31/24
Outputs
Target Audience:The results were disseminated through peer-reviewed publications and meeting presentations, including the Gordon Research Conference and CoFE. In addition, the PI also shared the results with industry members of companies through the campus venture catalyst group and events such as Natural Product Expo. Changes/Problems:There are delays due to the recruitment of postdoctoral scholars to support this project. We plan to submita no-cost extension request. What opportunities for training and professional development has the project provided?The project provided training opportunities to a graduate student, twopostdoctoral fellows, and one project scientist. In addition 4 undergraduate researchers have been engaged in research activities during this period. How have the results been disseminated to communities of interest?The research results were shared through scientific conferences, peer-reviewed publications, and engagement with industries through public events such as Natural product expo presentations by the PI. What do you plan to do during the next reporting period to accomplish the goals?The goals of the next phase are: 1. Complete the pending manuscripts from the year two studies 2. Assessment of the ability of 3D scaffolds and soluble oligosaccharides derived from black beans spent fractions to promote growth and mucoadhesion of probiotics in a simulated colonic environment 3. Optimization of the probiotic biofilm formation on plant-based scaffolds and evaluation of in-vivo persistence
Impacts
What was accomplished under these goals?
Novel food products rich in probiotics and prebiotics have significant potential to impact human health. However, the limited viability of probiotics during delivery to the gut and the lack of persistence of these bacteria in the gut significantly limits the success of these novel products. The harsh gastrointestinal conditions reduce the delivery of viable probiotics in food products, and the lack of persistence and growth is constrained by the poor mucoadhesion and availability of selective prebiotics, respectively. To address these challenges our interdisciplinary team is developing plant derived 3D scaffolds for the delivery of probiotics and enhance their persistence in-vivo. In this project period, our interdisciplinary team has made progress in the following goals: (a) Compositional analysis of probiotic biofilm in apple tissue: In our previous report we reported on our discovery of the unique potential of 3D scaffolds with probiotic biofilm to significantly enhance both the survivability and persistence of probiotic cells in-vitro and in-vivo. In this period, we have evaluated the composition of these probiotic biofilms using FTIR spectroscopy and LC-QQQ MS. The results of this analyses reveal significant increase in microbial biomass and associated extracellular biopolymer. The biopolymer fraction was significantly enriched in polysaccharide fraction and the protein fractions based on the results of IR measurement. For the LC-QQQ MS, the team has been optimizing the extraction and characterization process, and developing the analytical standards and the derivatization steps with 1-pheny-3-methyl-5-pyrazolone (PMP) methanolic solution for analysis of the carbohydrate composition of these biofilms. Fourteen monosaccharide standards were successfully resolved chromatographically in a 25 min binary gradient after derivatization, covering the full range of potential building blocks for the biopolymer. Mass spectrometry analysis revealed a varied composition, dominated by glucose, followed by galactose and xylose in comparable quantities, alongside with arabinose and mannose, and lastly, rhamnose and fucose found in lower abundances. (b) Mechanistic understanding of probiotic delivery and the role of structural properties: Many of the probiotic delivery approaches have been evaluated using empirical approaches. To develop a more fundamental understanding, our team has initiated mechanistic modeling of transport, persistence, and attachment of bacteria in the gut. The current model simulates the coupled movement of chyme and the mass transport of bacteria in the gut, including factors such as bulk mobility due to peristalsis and binding interactions with mucin and epithelial tissues. Imaging measurements were conducted to characterize the residual fractions or structure of the 3D scaffold in the gut. For this characterization, the samples of intestinal and cecal isolates from an animal study were imaged. The results reveal a distinct fiber-like morphology in the 3D scaffold in the small intestine, with distributed bacteria along the fiber length. In our next steps, we plan to evaluate the role of these fiber-like morphologies in influencing the distribution of the bacteria in the gut. (c) Optimization of extraction processes from beans and characterization of the spent fractions: One of the goals of this project is to utilize other by-products from the plant protein extraction industries to develop scaffolds for the delivery of probiotics. In this regard our team has been optimizing the extraction process from ground black beans and chickpeas. When these fractions were extracted using a conventional alkaline extraction method (AEP) widely used in the industry to obtain protein concentrates and/or isolates. The TPE of the black bean and chickpea extractions were 19.2 ± 1.5% and 25.3 ± 1.7%, demonstrating that a majority of the pulse proteins remained in the insoluble fraction. The poor extractability is likely due to the large particle size of the ground pulses used in this study. Comparatively, when black bean and chickpea flour were used and extraction was performed under the same conditions, protein extraction yields of 75% (black bean) and 63% (chickpea) were achieved. The residual fractions rich in carbohydrates and residual protein will be further evaluated to develop 3D scaffolds.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Nitin N, Plant Biomaterials for Pathogen Control and Promoting Delivery of Probiotics
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2024
Citation:
Biofilm scaffolds for the delivery of probiotics, COFE, 2024
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2024
Citation:
3d scaffolds for the delivery of probiotics, Gordon Research Conference, Lactic acid bacteria,
- Type:
Journal Articles
Status:
Published
Year Published:
2024
Citation:
1. Yang, J. S., Dias, F. F. G., Pham, T. T. K., Barile, D., & de Moura Bell, J. M. L. N. (2024). A sequential fractionation approach to understanding the physicochemical and functional properties of aqueous and enzyme-assisted aqueous extracted black bean proteins. Food Hydrocolloids, 146, 109250. https://doi.org/10.1016/j.foodhyd.2023.109250
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Progress 06/01/22 to 05/31/23
Outputs
Target Audience:The target audience includes food manufacturing andnutritional product companies. In addition, the target audience also includes researchers in academic and government laboratories. In this period, the target audiences reached by our efforts include both industry participants and researchers in academic and government laboratories. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The project provided training opportunities to a graduate student and a postdoctoral fellow. The postdoctoral fellow was promoted to project scientist in this review period also. How have the results been disseminated to communities of interest?The results were disseminated through peer-reviewed publications and presentations at meetings, including COFE and ACS meetings. In addition,the PI also shared the results with industry members of companies, including Kraft Heinz and Nestle, during their visit to the UCD campus. What do you plan to do during the next reporting period to accomplish the goals?Aim 2:Assessment of the ability of 3D scaffoldsand soluble oligosaccharides derived from apples and black beans spent fractions to promote growth and mucoadhesion of probiotics in a simulated colonic environment Aim 3: Evaluation ofin-vivopersistence and growth of probiotics delivered using the optimizedcompositions of 3Dscaffolds containing probiotics and oligosaccharides and their influence on high-fat diet-induced inflammation
Impacts
What was accomplished under these goals?
Assessment of the enhanced survivability of probiotics during simulatedin vitrodigestion using 3D scaffolds derived from apple and black beans extraction byproducts and also promote growth of bacteria in-vitro. In the first year, we focused on evaluating the enhanced survivability of probiotics using 3D scaffolds derived from plant by- products. Our first set of efforts was focused on Apple by-products. This study develops a novel low-cost microbial delivery system by transforming common food materials such as apple tissue into a 3D scaffold. Apple tissue scaffold was constructed by decellularization of intact tissue using a minimal amount of sodium dodecyl sulfate (0.5 % w/v). Vacuum-assisted infusion of model probiotic Lactobacillus cells led to a high encapsulation yield of probiotic cells (1010 CFU/g of scaffold) in 3D scaffolds on a wet basis. The bio-polymer coated 3D scaffolds with infused cells significantly enhanced the survivability of infused probiotic cells during simulated gastric and intestinal digestions. In addition, imaging and plate counting results validate the growth of the infused cells in the 3D scaffold after 1-2 days of fermentation in MRS media, while cells without infusion in the scaffold had limited attachment with the intact apple tissue. Overall, these results highlight the potential of the apple tissue-derived 3D scaffold to deliver probiotic cells and include the biochemical compositions to support the growth of delivered microbial cells in the colon.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2023
Citation:
Apple-derived 3D scaffold for improving gastrointestinal viability and in-situ growth of probiotics
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
Plant derived compositions for improved delivery and persistence of probiotics in the gut, COFE, 2022
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