De-Novo Design of Antimicrobial Peptides for Plant Protection

Abstract

This work describes the de-novo design of peptides that inhibit a broad range of plant pathogens. Four structurally different groups of peptides were developed that differ in size and position of their charged and hydrophobic clusters and were assayed for their ability to inhibit bacterial growth and fungal spore germination. Several peptides are highly active at concentrations between 0,1 and 1 µg/ml against plant pathogenic bacteria, such as Pseudomonas syringae, Pectobacterium carotovorum, and Xanthomonas vesicatoria. Importantly, no hemolytic activity could be detected for these peptides at concentrations up to 200 µg/ml. Moreover, the peptides are also active after spraying on the plant surface demonstrating a possible way of application. In sum, our designed peptides represent new antimicrobial agents and with the increasing demand for antimicrobial compounds for production of “healthy” food, these peptides might serve as templates for novel antibacterial and antifungal agents.

Comparative analysis of internalisation, haemolytic, cytotoxic and antibacterial effect of membrane-active cationic peptides: aspects of experimental setup

“…Important variants are as follows: (i) concentration of RBC (0.5% v/v (Hollmann et al 2016), 1% (Kobayashi et al 2000), 2% (Yang et al 2013), 4% (Lee and Lee 2008;Song et al 2005), 20% (Chongsiriwatana et al 2008), 2 × 10 6 cells/mL (Li et al 2005) or 2.5 × 10 8 cells/mL (Dathe et al 2001;Dathe et al 1996), 1.2 × 10 9 cells/mL (Zeitler et al 2013) etc. ); (ii) incubation media (PBS (Chongsiriwatana et al 2008;Dennison and Phoenix 2014;Hollmann et al 2016;Lee and Lee 2008;Song et al 2005), Tris (Dathe et al 2001;Dathe et al 1996;Zeitler et al 2013), RPMI-1640 or HEPES buffer (Li et al 2005), 0.9% NaCl or normal saline (Wu et al 2014), etc.…”

“…Important variants are as follows: (i) concentration of RBC (0.5% v/v (Hollmann et al 2016), 1% (Kobayashi et al 2000), 2% (Yang et al 2013), 4% (Lee and Lee 2008;Song et al 2005), 20% (Chongsiriwatana et al 2008), 2 × 10 6 cells/mL (Li et al 2005) or 2.5 × 10 8 cells/mL (Dathe et al 2001;Dathe et al 1996), 1.2 × 10 9 cells/mL (Zeitler et al 2013) etc. ); (ii) incubation media (PBS (Chongsiriwatana et al 2008;Dennison and Phoenix 2014;Hollmann et al 2016;Lee and Lee 2008;Song et al 2005), Tris (Dathe et al 2001;Dathe et al 1996;Zeitler et al 2013), RPMI-1640 or HEPES buffer (Li et al 2005), 0.9% NaCl or normal saline (Wu et al 2014), etc. ); (iii) incubation time (30 min (Hollmann et al 2016;Wu et al 2014), 1 hour (Chongsiriwatana et al 2008;Dennison and Phoenix 2014;Yang et al 2013), 4 hrs (Li et al 2005) or 18-24 hrs (Mojsoska et al 2015)); (iii) used positive control (0.1% Triton-X (Dennison and Phoenix 2014;Lee and Lee 2008;Song et al 2005), 1% Triton-X (Mojsoska et al 2015), 10% Triton-X (Wu et al 2014), distilled water (Hollmann et al 2016;Yang et al 2013), 2 % SDS (Zeitler et al 2013), 0.05% saponin (Davanco et al 2014).…”

The PDB database is a rich source of alpha-helical anti-microbial peptides to combat disease causing pathogens

“…Genetic approaches aim at achieving the same goal of enhancing immunity through rational design of peptides 13, 47 , which are then incorporated into the genome 29, 31, 48 . Also, it is important to ensure that these non-endogenous genomic fragments have minimal effect on humans for their commercial viability 32 .…”

“…Not surprisingly, these AHs are the targets for antibody binding 7, 8 and therapeutic agents 9 . These therapies in turn use AH peptides against both viral 10– 12 and bacterial pathogens 13 .…”

Anti-proliferative effect on a colon adenocarcinoma cell line exerted by a membrane disrupting antimicrobial peptide KL15

“…2,14 Since the structure, cationic and hydrophobic characteristics of an AMP may determine its mode of action, direct modification of these features allows the rational design of potent AMPs. 15 Recently, we have identified several novel bacteriocin loci from the genomic sequences of a completely sequenced lactic acid bacteria (LAB) Lactobacillus (L.) casei ATCC 334 (GI number: 116103724) which is otherwise not known as a bacteriocin producer. 16 The DNA sequences of some of these putative AMPs such as m2163 and m2386 were cloned and heterologously expressed.…”