# Engineering Antimicrobial Polypeptides against Pathogenic Bacteria

> **NIH NIH R01** · UNIVERSITY OF MINNESOTA · 2020 · $286,945

## Abstract

Project Summary
Antibiotic-resistant microbial infections are a critical healthcare problem in the U.S. and the world. Improved
antimicrobial strategies – and an efficient mechanism for their ongoing generation to keep pace with microbial
evolution – are sorely needed. Vancomycin-resistant Enterococcus faecium is particularly problematic and is in
the CDC's “Serious Threat” category. Antimicrobial polypeptides (AMPs) offer a compelling class of
therapeutics to provide potency, specificity, and hindrance to resistance. Engineered probiotic lactic acid
bacteria can provide local delivery of AMPs to overcome barriers to traditional therapeutic delivery. The
objective of the proposed studies is to engineer AMPs enterocin A and endolysin ORF9, via an innovative
directed evolution platform, for enhanced potency and specificity to combat E. faecium infections. The AMPs
will be engineered and evaluated via secretion from two probiotics: Lactococcus lactis and Lactobacillus
johnsonii. The objective will be achieved via three aims. (1) Develop a platform for high throughput (~107/day),
quantitative selection of potent AMPs as secreted from probiotics. This will be achieved via microdroplet co-
encapsulation of probiotic and pathogen along with fluorescence-activated cell sorting. (2) Elucidate efficient
evolutionary pathways for AMPs ORF9 lysin and entA to enhance potency and specificity, from a probiotic
host, against E. faecium. Innovative combinatorial library designs, including some informed by bioinformatics,
will be used and compared. (3) Evaluate engineered probiotic/AMP efficacy against E. faecium in a murine
model while elucidating the impacts of potency, host, secretion efficiency, dose, and AMP diversity on E.
faecium inhibition and resistance reduction.

## Key facts

- **NIH application ID:** 9840488
- **Project number:** 5R01GM121777-04
- **Recipient organization:** UNIVERSITY OF MINNESOTA
- **Principal Investigator:** Benjamin Hackel
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $286,945
- **Award type:** 5
- **Project period:** 2017-01-01 → 2021-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9840488

## Citation

> US National Institutes of Health, RePORTER application 9840488, Engineering Antimicrobial Polypeptides against Pathogenic Bacteria (5R01GM121777-04). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/9840488. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*