Recipient Organization
HOOFPRINT BIOME, INC.
522 BERWICK VALLEY LN
CARY,NC 27513
Performing Department
(N/A)
Non Technical Summary
Methanogenic microbes in cattle rumen are responsible for nearly 10% of global greenhouse gas emissions. This methane production isnot vital to the cow's health, and can consume up to 12% of an animal'sfeed energy. Existing methane reduction products in development either lack scalability or are prohibitively expensive. In contrast, Hoofprint Biome is innovating a solution that increases cattle profitability and eliminates methane.Our groundbreaking discovery is a library ofnatural, bioactive peptides that putatively inhibit methane emissions from the microbes in a cow'srumen. We propose to ultimately secrete these peptides in the rumen using a probiotic yeast, Saccharomyces boulardii, which has already been proven to increase milk yield and feed efficiency by over 5%. This proposal aims to screen and identify the most potent and consistentpeptides from our library and engineer their secretion fromS. boulardii.The successful completion of this phase I proposal will yield a scalable methane-reducingprobiotic ready for field trials. The phase II proposal will support milk yield, feed efficiency, and methane reduction trials from cattle. If successful, our product could enhance dairy and beef profitability by over 25%, and and play an essential role in the US hitting 1.5 °C emissions reduction targets.
Animal Health Component
40%
Research Effort Categories
Basic
10%
Applied
40%
Developmental
50%
Goals / Objectives
In this USDA SBIR Phase I effort, Hoofprint proposes to develop bioactive peptides to eliminate methane emissions from cattle and to secrete them from the probiotic yeast, S. boulardii. We have computationally identified a library of novel bioactive peptides, and the goals of this proposal are to use this library to develop acost-effective probiotic feed additiveproduct. Our technical objectives are:Objective A. Screen and selectanti-methanepeptides in vitro.Objective B. Demonstrate methane reduction using anti-methane peptide secretion by S. boulardiiHoofprint plans to answer to following technical questions to establish the technical feasibility of the proposed approach:Which of our identified peptides has the strongest reduction of methane production? Is secretion of multiple peptides required for complete methane elimination? What is the final concentration of peptides secreted from S. boulardii? What will be the expected effective dose in vivo?Project success requires: (1) identification of bioactive peptides that can reduce methane by >80% using our in vitro articial rumen culturesystem; (2) secretion of peptidesfrom Hoofprint's S. boulardii platform in sufficient quantities for a biologically relevant effectand reducing methane from in vitro cultures by at least 80%. To carry out these objectives, we will leverage Hoofprint's prior experience with cell-free peptide synthesis,artificial rumen (in vitro) fermentation, and probiotic yeast engineering. The successful completion of these objectives will yield a product that will decrease methane from the cattle rumen microbiome by at minimum 80% in vitro, while simultaneously improving the health and productivity of cattle. The objectives of the Phase II proposal will be to translate these findings toin vivo trials for methane reduction, feed efficiency, and animal health.
Project Methods
Hoofprint has established a high throughput platform for testing rumen fluid. Microbes and rumen fluid are directly removed from cannulated cattle and mixed with buffer and feed substrate. The team has previously used this model for testing compounds for methane inhibition, assessing the digestive efficiency of various feeds, and assessing microbial populations. Methane inhibition within these in-vitro systems is highly correlated with in-vivo behaviors. In order to test the efficacy of our bioactive peptides, we will express the peptides using cell-free protein synthesis. We will then add peptides to our in vitro rumen fluid systems to assess impact of peptides on microbial dynamics and methane production. Once we have identified peptide(s) that significantly and consistently reduce rumen methane in vitro, we will then engineer the secretion of these peptides using probiotics. The goal is to express high levels of these bioactive peptides for a low cost by using probiotic yeast. Expression vectors will be used to express the enzymes, enhance expression,andquantify the amount of peptide generated before adding the yeast-expressed peptides to the rumen fluid culture system to quantify methane emissions. Methane will be measured by headspace in the rumen fluid culture in addition to microbial gene expression and VFA profiles from the culture.