Location: Functional Foods Research
Project Number: 5010-41000-182-004-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Sep 1, 2023
End Date: Dec 31, 2024
Objective:
1. Evaluate the gastrointestinal digestibility and bio-accessibility of alfalfa proteins and phytochemicals.
2. Determine the impact of alfalfa dietary fibers on the human fecal microbiome.
Approach:
Proteins and dietary fibers will be extracted from alfalfa seeds and In vitro digestibility and bioaccessibility, Gastrointestinal toxicity, and In vitro fermentation will be conducted by Cooperator. Specifically,
1) In vitro gastrointestinal (GI) digestibility and bio-accessibility of proteins will be performed following an industry standard digestion protocol. A manual mincer to simulate the chewing process is used, and then the minced samples will be transferred into a stomacher bag, mixed with simulated salivary fluid and a-amylase and processed. Then, the solution will be centrifuged at 10,000g for 30 minutes at 4°C to separate the digested soluble fraction.
2) The GI-digested soluble fraction from in vitro digestion will be used to evaluate the toxicity and in-vitro bioavailability of the short peptides and amino acids.
3) The dietary fiber fractions from alfalfa seeds will be subjected to in vitro digestion and dialysis to remove residual digestible material. Each alfalfa fraction recovered after in vitro digestion will be mixed with fecal bacteria collected from healthy volunteers under strict anaerobic conditions and fermented for up to 96 hours. Following fermentation, changes in the microbiota composition will be assessed using amplicon sequencing of the 16S rRNA gene.
