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Biochemical properties and yields of diverse bacterial laccase-like multicopper oxidases expressed in Escherichia coli
Laccases are multi-copper oxidases that oxidize a broad range of substrates at the expense of molecular oxygen, without any need for co-factor regeneration. These enzymes bear high potential for the sustainable synthesis of fine chemicals and the modification of (bio)polymers. Here we describe cloni...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4464401/ https://www.ncbi.nlm.nih.gov/pubmed/26068013 http://dx.doi.org/10.1038/srep10465 |
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author | Ihssen, Julian Reiss, Renate Luchsinger, Ronny Thöny-Meyer, Linda Richter, Michael |
author_facet | Ihssen, Julian Reiss, Renate Luchsinger, Ronny Thöny-Meyer, Linda Richter, Michael |
author_sort | Ihssen, Julian |
collection | PubMed |
description | Laccases are multi-copper oxidases that oxidize a broad range of substrates at the expense of molecular oxygen, without any need for co-factor regeneration. These enzymes bear high potential for the sustainable synthesis of fine chemicals and the modification of (bio)polymers. Here we describe cloning and expression of five novel bacterial laccase-like multi copper oxidases (LMCOs) of diverse origin which were identified by homology searches in online databases. Activity yields under different expression conditions and temperature stabilities were compared to three previously described enzymes from Bacillus subtilis, Bacillus pumilus and Bacillus clausii. In almost all cases, a switch to oxygen-limited growth conditions after induction increased volumetric activity considerably. For proteins with predicted signal peptides for secretion, recombinant expression with and without signal sequence was investigated. Bacillus CotA-type LMCOs outperformed enzymes from Streptomyces and Gram-negative bacteria with respect to activity yields in Escherichia coli and application relevant biochemical properties. The novel Bacillus coagulans LMCO combined high activity yields in E. coli with unprecedented activity at strong alkaline pH and high storage stability, making it a promising candidate for further development. |
format | Online Article Text |
id | pubmed-4464401 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-44644012015-06-18 Biochemical properties and yields of diverse bacterial laccase-like multicopper oxidases expressed in Escherichia coli Ihssen, Julian Reiss, Renate Luchsinger, Ronny Thöny-Meyer, Linda Richter, Michael Sci Rep Article Laccases are multi-copper oxidases that oxidize a broad range of substrates at the expense of molecular oxygen, without any need for co-factor regeneration. These enzymes bear high potential for the sustainable synthesis of fine chemicals and the modification of (bio)polymers. Here we describe cloning and expression of five novel bacterial laccase-like multi copper oxidases (LMCOs) of diverse origin which were identified by homology searches in online databases. Activity yields under different expression conditions and temperature stabilities were compared to three previously described enzymes from Bacillus subtilis, Bacillus pumilus and Bacillus clausii. In almost all cases, a switch to oxygen-limited growth conditions after induction increased volumetric activity considerably. For proteins with predicted signal peptides for secretion, recombinant expression with and without signal sequence was investigated. Bacillus CotA-type LMCOs outperformed enzymes from Streptomyces and Gram-negative bacteria with respect to activity yields in Escherichia coli and application relevant biochemical properties. The novel Bacillus coagulans LMCO combined high activity yields in E. coli with unprecedented activity at strong alkaline pH and high storage stability, making it a promising candidate for further development. Nature Publishing Group 2015-06-12 /pmc/articles/PMC4464401/ /pubmed/26068013 http://dx.doi.org/10.1038/srep10465 Text en Copyright © 2015, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Ihssen, Julian Reiss, Renate Luchsinger, Ronny Thöny-Meyer, Linda Richter, Michael Biochemical properties and yields of diverse bacterial laccase-like multicopper oxidases expressed in Escherichia coli |
title | Biochemical properties and yields of diverse bacterial laccase-like multicopper oxidases expressed in Escherichia coli |
title_full | Biochemical properties and yields of diverse bacterial laccase-like multicopper oxidases expressed in Escherichia coli |
title_fullStr | Biochemical properties and yields of diverse bacterial laccase-like multicopper oxidases expressed in Escherichia coli |
title_full_unstemmed | Biochemical properties and yields of diverse bacterial laccase-like multicopper oxidases expressed in Escherichia coli |
title_short | Biochemical properties and yields of diverse bacterial laccase-like multicopper oxidases expressed in Escherichia coli |
title_sort | biochemical properties and yields of diverse bacterial laccase-like multicopper oxidases expressed in escherichia coli |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4464401/ https://www.ncbi.nlm.nih.gov/pubmed/26068013 http://dx.doi.org/10.1038/srep10465 |
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