A new generation of recombinant polypeptides combines multiple protein domains for effective antimicrobial activity
Abstract Background Although most of antimicrobial peptides (AMPs), being relatively short, are produced by chemical synthesis, several AMPs have been produced using recombinant technology. However, AMPs could be cytotoxic to the producer cell, and if small they can be easily degraded. The objective...
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doaj-272593b8b738450aaf644320727ef24f2020-11-25T03:26:19ZengBMCMicrobial Cell Factories1475-28592020-06-011911710.1186/s12934-020-01380-7A new generation of recombinant polypeptides combines multiple protein domains for effective antimicrobial activityRamon Roca-Pinilla0Adrià López-Cano1Cristina Saubi2Elena Garcia-Fruitós3Anna Arís4Department of Ruminant Production, Institute of Agriculture and Food Research (IRTA)Department of Ruminant Production, Institute of Agriculture and Food Research (IRTA)Department of Ruminant Production, Institute of Agriculture and Food Research (IRTA)Department of Ruminant Production, Institute of Agriculture and Food Research (IRTA)Department of Ruminant Production, Institute of Agriculture and Food Research (IRTA)Abstract Background Although most of antimicrobial peptides (AMPs), being relatively short, are produced by chemical synthesis, several AMPs have been produced using recombinant technology. However, AMPs could be cytotoxic to the producer cell, and if small they can be easily degraded. The objective of this study was to produce a multidomain antimicrobial protein based on recombinant protein nanoclusters to increase the yield, stability and effectivity. Results A single antimicrobial polypeptide JAMF1 that combines three functional domains based on human α-defensin-5, human XII-A secreted phospholipase A2 (sPLA2), and a gelsolin-based bacterial-binding domain along with two aggregation-seeding domains based on leucine zippers was successfully produced with no toxic effects for the producer cell and mainly in a nanocluster structure. Both, the nanocluster and solubilized format of the protein showed a clear antimicrobial effect against a broad spectrum of Gram-negative and Gram-positive bacteria, including multi-resistant strains, with an optimal concentration between 1 and 10 µM. Conclusions Our findings demonstrated that multidomain antimicrobial proteins forming nanoclusters can be efficiently produced in recombinant bacteria, being a novel and valuable strategy to create a versatile, highly stable and easily editable multidomain constructs with a broad-spectrum antimicrobial activity in both soluble and nanostructured format.http://link.springer.com/article/10.1186/s12934-020-01380-7Antimicrobial peptidesAntimicrobial resistanceInclusion bodiesMultidomain proteinSolubilizationRecombinant production |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Ramon Roca-Pinilla Adrià López-Cano Cristina Saubi Elena Garcia-Fruitós Anna Arís |
spellingShingle |
Ramon Roca-Pinilla Adrià López-Cano Cristina Saubi Elena Garcia-Fruitós Anna Arís A new generation of recombinant polypeptides combines multiple protein domains for effective antimicrobial activity Microbial Cell Factories Antimicrobial peptides Antimicrobial resistance Inclusion bodies Multidomain protein Solubilization Recombinant production |
author_facet |
Ramon Roca-Pinilla Adrià López-Cano Cristina Saubi Elena Garcia-Fruitós Anna Arís |
author_sort |
Ramon Roca-Pinilla |
title |
A new generation of recombinant polypeptides combines multiple protein domains for effective antimicrobial activity |
title_short |
A new generation of recombinant polypeptides combines multiple protein domains for effective antimicrobial activity |
title_full |
A new generation of recombinant polypeptides combines multiple protein domains for effective antimicrobial activity |
title_fullStr |
A new generation of recombinant polypeptides combines multiple protein domains for effective antimicrobial activity |
title_full_unstemmed |
A new generation of recombinant polypeptides combines multiple protein domains for effective antimicrobial activity |
title_sort |
new generation of recombinant polypeptides combines multiple protein domains for effective antimicrobial activity |
publisher |
BMC |
series |
Microbial Cell Factories |
issn |
1475-2859 |
publishDate |
2020-06-01 |
description |
Abstract Background Although most of antimicrobial peptides (AMPs), being relatively short, are produced by chemical synthesis, several AMPs have been produced using recombinant technology. However, AMPs could be cytotoxic to the producer cell, and if small they can be easily degraded. The objective of this study was to produce a multidomain antimicrobial protein based on recombinant protein nanoclusters to increase the yield, stability and effectivity. Results A single antimicrobial polypeptide JAMF1 that combines three functional domains based on human α-defensin-5, human XII-A secreted phospholipase A2 (sPLA2), and a gelsolin-based bacterial-binding domain along with two aggregation-seeding domains based on leucine zippers was successfully produced with no toxic effects for the producer cell and mainly in a nanocluster structure. Both, the nanocluster and solubilized format of the protein showed a clear antimicrobial effect against a broad spectrum of Gram-negative and Gram-positive bacteria, including multi-resistant strains, with an optimal concentration between 1 and 10 µM. Conclusions Our findings demonstrated that multidomain antimicrobial proteins forming nanoclusters can be efficiently produced in recombinant bacteria, being a novel and valuable strategy to create a versatile, highly stable and easily editable multidomain constructs with a broad-spectrum antimicrobial activity in both soluble and nanostructured format. |
topic |
Antimicrobial peptides Antimicrobial resistance Inclusion bodies Multidomain protein Solubilization Recombinant production |
url |
http://link.springer.com/article/10.1186/s12934-020-01380-7 |
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