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Functional Expression of GFP-Fused Class I Lanthipeptides in Escherichia coli

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논문

Functional Expression of GFP-Fused Class I Lanthipeptides in Escherichia coli

학술지

ACS Synthetic biology

저자명

Van Staden, Anton Du Preez; Faure, Lindsay M.; Vermeulen, Ross R.; Dicks, Leon M. T.; Smith, Carine

초록

<P>Lanthipeptides are ribosomally synthesized and post-translationally modified peptides, with several having antimicrobial activity. The biosynthetic machinery responsible for modification of the class I lanthipeptide nisin provides a means for modification of a diverse range of lanthipeptides. However, literature regarding expression of class I lanthipeptides in a malleable Gram-negative host such as <I>Escherichia coli</I> is limited. Here, we coexpressed precursor class I lanthipeptides fused to green fluorescent protein (GFP) along with the dehydratase and cyclase from the nisin operon. Fusion to GFP did not interfere with post-translational modifications as antimicrobially active nisin could be proteolytically liberated from the expressed GFP fusion. Additionally, we used this system to express two other class I lanthipeptides precursors fused to GFP (Pep5 and epilancin 15X), although only Pep5 exhibited consistent antimicrobial activity. This is the first report of a GFP-based fusion expression system for the expression of class I lanthipeptides in <I>E. coli</I>. The GFP-based fusion expression system is a robust system with the advantage of directly visualizing expression and purification through GFP fluorescence.</P><BR>[FIG OMISSION]</BR>

발행연도

2019

발행기관

American Chemical Society

ISSN

2161-5063

8

10

페이지

pp.2220-2227

주제어

nisin; class I lanthipeptides; Escherichia coli heterologous expression; GFP-fusion; protein engineering

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1 2023-12-11

논문; 2019-12-31

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