« Previous Abstract"Modeling the plant uptake of organic chemicals, including the soil-air-plant pathway"    Next AbstractApplication of a Microfluidic Gas-to-Liquid Interface for Extraction of Target Amphetamines and Precursors from Air Samples »

Biochemistry

Title:		Design of a De Novo Aggregating Antimicrobial Peptide and a Bacterial Conjugation-Based Delivery System
Author(s):		Collins LT; Otoupal PB; Campos JK; Courtney CM; Chatterjee A;
Address:		"Department of Chemical and Biological Engineering , University of Colorado , Boulder , Colorado 80303 , United States"
Journal Title:		Biochemistry
Year:		2019
Volume:		20181109
Issue:		11
Page Number:		1521 - 1526
DOI:		10.1021/acs.biochem.8b00888
ISSN/ISBN:		1520-4995 (Electronic) 0006-2960 (Linking)
Abstract:		"Antibacterial resistance necessitates the development of novel treatment methods for infections. Protein aggregates have recently been applied as antimicrobials to disrupt bacterial homeostasis. Past work on protein aggregates has focused on genome mining for aggregation-prone sequences in bacterial genomes rather than on rational design of aggregating antimicrobial peptides. Here, we use a synthetic biology approach to design an artificial gene encoding a de novo aggregating antimicrobial peptide. This artificial gene, opaL (overexpressed protein aggregator lipophilic), disrupts bacterial homeostasis by expressing extremely hydrophobic peptides. When this hydrophobic sequence is disrupted by acidic residues, consequent aggregation and antimicrobial effect decrease. Further, we developed a probiotic delivery system using the broad-host range conjugative plasmid RK2 to transfer the gene from donor to recipient bacteria. We utilize RK2 to mobilize a shuttle plasmid carrying opaL by adding the RK2 origin of transfer. We show that opaL is nontoxic to the donor, allowing for maintenance and transfer since its expression is under control of a promoter with a recipient-specific T7 RNA polymerase. Upon mating of donor and recipient Escherichia coli, we observe selective growth repression in T7 polymerase-expressing recipients. This technique could be used to target desired pathogens by selecting pathogen-specific promoters to control T7 RNA polymerase expression and provides a basis for the design and delivery of aggregating antimicrobial peptides"
Keywords:		"Anti-Bacterial Agents/*chemical synthesis/*pharmacology Bacteria/metabolism Bacterial Proteins/metabolism Conjugation, Genetic/genetics Drug Resistance, Bacterial/drug effects Escherichia coli/metabolism Escherichia coli Proteins/metabolism Gene Expressio;"
Notes:		"MedlineCollins, Logan T Otoupal, Peter B Campos, Jocelyn K Courtney, Colleen M Chatterjee, Anushree eng Research Support, U.S. Gov't, Non-P.H.S. 2018/11/08 Biochemistry. 2019 Mar 19; 58(11):1521-1526. doi: 10.1021/acs.biochem.8b00888. Epub 2018 Nov 9"