Identification of ε-Poly-L-lysine as an Antimicrobial Product from an Epichloë Endophyte and Isolation of Fungal ε-PL Synthetase Gene

The endophytic fungus Epichloë festucae is known to produce bioactive metabolites, which consequently protect the host plants from biotic and abiotic stresses. We previously found that the overexpression of vibA (a gene for transcription factor) in <...

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Main Authors:	Enkhee Purev, Tatsuhiko Kondo, Daigo Takemoto, Jennifer T. Niones, Makoto Ojika
Format:	Article
Language:	English
Published:	MDPI AG 2020-02-01
Series:	Molecules
Subjects:	ε-polylysine <i>epichloë</i> endophyte fungicide transcription factor
Online Access:	https://www.mdpi.com/1420-3049/25/5/1032

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spelling	doaj-545bb8a9444c4b489a0fdd2f609b925c2020-11-25T00:31:47ZengMDPI AGMolecules1420-30492020-02-01255103210.3390/molecules25051032molecules25051032Identification of ε-Poly-L-lysine as an Antimicrobial Product from an <i>Epichloë</i> Endophyte and Isolation of Fungal ε-PL Synthetase GeneEnkhee Purev0Tatsuhiko Kondo1Daigo Takemoto2Jennifer T. Niones3Makoto Ojika4Department of Applied Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa-ku, Nagoya 464-8601, JapanDepartment of Applied Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa-ku, Nagoya 464-8601, JapanDepartment of Plant Production Sciences, Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa-ku, Nagoya 464-8601, JapanDepartment of Plant Production Sciences, Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa-ku, Nagoya 464-8601, JapanDepartment of Applied Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa-ku, Nagoya 464-8601, JapanThe endophytic fungus <i>Epichloë festucae</i> is known to produce bioactive metabolites, which consequently protect the host plants from biotic and abiotic stresses. We previously found that the overexpression of <i>vibA</i> (a gene for transcription factor) in <i>E. festucae</i> strain E437 resulted in the secretion of an unknown fungicide. In the present study, the active substance was purified and chemically identified as ε-poly-L-lysine (ε-PL), which consisted of 28−34 lysine units. The productivity was 3.7-fold compared with that of the wild type strain E437. The isolated ε-PL showed inhibitory activity against the spore germination of the plant pathogens <i>Drechslera erythrospila</i>, <i>Botrytis cinerea</i>, and <i>Phytophthora infestans</i> at 1−10 μg/mL. We also isolated the fungal gene “<i>epls</i>” encoding ε-PL synthetase Epls. Overexpression of <i>epls</i> in the wild type strain E437 resulted in the enhanced production of ε-PL by 6.7-fold. Interestingly, overexpression of <i>epls</i> in the different strain <i>E. festucae</i> Fl1 resulted in the production of shorter ε-PL with 8−20 lysine, which exhibited a comparable antifungal activity to the longer one. The results demonstrate the first example of ε-PL synthetase gene from the eukaryotic genomes and suggest the potential of enhanced expression of <i>vibA</i> or/and <i>epls</i> genes in the <i>Epichloë</i> endophyte for constructing pest-tolerant plants.https://www.mdpi.com/1420-3049/25/5/1032ε-polylysine<i>epichloë</i>endophytefungicidetranscription factor
collection	DOAJ
language	English
format	Article
sources	DOAJ
author	Enkhee Purev Tatsuhiko Kondo Daigo Takemoto Jennifer T. Niones Makoto Ojika
spellingShingle	Enkhee Purev Tatsuhiko Kondo Daigo Takemoto Jennifer T. Niones Makoto Ojika Identification of ε-Poly-L-lysine as an Antimicrobial Product from an <i>Epichloë</i> Endophyte and Isolation of Fungal ε-PL Synthetase Gene Molecules ε-polylysine <i>epichloë</i> endophyte fungicide transcription factor
author_facet	Enkhee Purev Tatsuhiko Kondo Daigo Takemoto Jennifer T. Niones Makoto Ojika
author_sort	Enkhee Purev
title	Identification of ε-Poly-L-lysine as an Antimicrobial Product from an <i>Epichloë</i> Endophyte and Isolation of Fungal ε-PL Synthetase Gene
title_short	Identification of ε-Poly-L-lysine as an Antimicrobial Product from an <i>Epichloë</i> Endophyte and Isolation of Fungal ε-PL Synthetase Gene
title_full	Identification of ε-Poly-L-lysine as an Antimicrobial Product from an <i>Epichloë</i> Endophyte and Isolation of Fungal ε-PL Synthetase Gene
title_fullStr	Identification of ε-Poly-L-lysine as an Antimicrobial Product from an <i>Epichloë</i> Endophyte and Isolation of Fungal ε-PL Synthetase Gene
title_full_unstemmed	Identification of ε-Poly-L-lysine as an Antimicrobial Product from an <i>Epichloë</i> Endophyte and Isolation of Fungal ε-PL Synthetase Gene
title_sort	identification of ε-poly-l-lysine as an antimicrobial product from an <i>epichloë</i> endophyte and isolation of fungal ε-pl synthetase gene
publisher	MDPI AG
series	Molecules
issn	1420-3049
publishDate	2020-02-01
description	The endophytic fungus <i>Epichloë festucae</i> is known to produce bioactive metabolites, which consequently protect the host plants from biotic and abiotic stresses. We previously found that the overexpression of <i>vibA</i> (a gene for transcription factor) in <i>E. festucae</i> strain E437 resulted in the secretion of an unknown fungicide. In the present study, the active substance was purified and chemically identified as ε-poly-L-lysine (ε-PL), which consisted of 28−34 lysine units. The productivity was 3.7-fold compared with that of the wild type strain E437. The isolated ε-PL showed inhibitory activity against the spore germination of the plant pathogens <i>Drechslera erythrospila</i>, <i>Botrytis cinerea</i>, and <i>Phytophthora infestans</i> at 1−10 μg/mL. We also isolated the fungal gene “<i>epls</i>” encoding ε-PL synthetase Epls. Overexpression of <i>epls</i> in the wild type strain E437 resulted in the enhanced production of ε-PL by 6.7-fold. Interestingly, overexpression of <i>epls</i> in the different strain <i>E. festucae</i> Fl1 resulted in the production of shorter ε-PL with 8−20 lysine, which exhibited a comparable antifungal activity to the longer one. The results demonstrate the first example of ε-PL synthetase gene from the eukaryotic genomes and suggest the potential of enhanced expression of <i>vibA</i> or/and <i>epls</i> genes in the <i>Epichloë</i> endophyte for constructing pest-tolerant plants.
topic	ε-polylysine <i>epichloë</i> endophyte fungicide transcription factor
url	https://www.mdpi.com/1420-3049/25/5/1032
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Identification of ε-Poly-L-lysine as an Antimicrobial Product from an <i>Epichloë</i> Endophyte and Isolation of Fungal ε-PL Synthetase Gene

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