A novel programmable lysozyme-based lysis system in Pseudomonas putida for biopolymer production

Abstract Cell lysis is crucial for the microbial production of industrial fatty acids, proteins, biofuels, and biopolymers. In this work, we developed a novel programmable lysis system based on the heterologous expression of lysozyme. The inducible lytic system was tested in two Gram-negative bacter...

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Autores principales:	José Manuel Borrero-de Acuña, Cristian Hidalgo-Dumont, Nicolás Pacheco, Alex Cabrera, Ignacio Poblete-Castro
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1be251829cf2424999c089d62f752eda
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spelling	oai:doaj.org-article:1be251829cf2424999c089d62f752eda2021-12-02T11:52:58ZA novel programmable lysozyme-based lysis system in Pseudomonas putida for biopolymer production10.1038/s41598-017-04741-22045-2322https://doaj.org/article/1be251829cf2424999c089d62f752eda2017-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-04741-2https://doaj.org/toc/2045-2322Abstract Cell lysis is crucial for the microbial production of industrial fatty acids, proteins, biofuels, and biopolymers. In this work, we developed a novel programmable lysis system based on the heterologous expression of lysozyme. The inducible lytic system was tested in two Gram-negative bacterial strains, namely Escherichia coli and Pseudomonas putida KT2440. Before induction, the lytic system did not significantly arrest essential physiological parameters in the recombinant E. coli (ECPi) and P. putida (JBOi) strain such as specific growth rate and biomass yield under standard growth conditions. A different scenario was observed in the recombinant JBOi strain when subjected to PHA-producing conditions, where biomass production was reduced by 25% but the mcl-PHA content was maintained at about 30% of the cell dry weight. Importantly, the genetic construct worked well under PHA-producing conditions (nitrogen-limiting phase), where more than 95% of the cell population presented membrane disruption 16 h post induction, with 75% of the total synthesized biopolymer recovered at the end of the fermentation period. In conclusion, this new lysis system circumvents traditional, costly mechanical and enzymatic cell-disrupting procedures.José Manuel Borrero-de AcuñaCristian Hidalgo-DumontNicolás PachecoAlex CabreraIgnacio Poblete-CastroNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q José Manuel Borrero-de Acuña Cristian Hidalgo-Dumont Nicolás Pacheco Alex Cabrera Ignacio Poblete-Castro A novel programmable lysozyme-based lysis system in Pseudomonas putida for biopolymer production
description	Abstract Cell lysis is crucial for the microbial production of industrial fatty acids, proteins, biofuels, and biopolymers. In this work, we developed a novel programmable lysis system based on the heterologous expression of lysozyme. The inducible lytic system was tested in two Gram-negative bacterial strains, namely Escherichia coli and Pseudomonas putida KT2440. Before induction, the lytic system did not significantly arrest essential physiological parameters in the recombinant E. coli (ECPi) and P. putida (JBOi) strain such as specific growth rate and biomass yield under standard growth conditions. A different scenario was observed in the recombinant JBOi strain when subjected to PHA-producing conditions, where biomass production was reduced by 25% but the mcl-PHA content was maintained at about 30% of the cell dry weight. Importantly, the genetic construct worked well under PHA-producing conditions (nitrogen-limiting phase), where more than 95% of the cell population presented membrane disruption 16 h post induction, with 75% of the total synthesized biopolymer recovered at the end of the fermentation period. In conclusion, this new lysis system circumvents traditional, costly mechanical and enzymatic cell-disrupting procedures.
format	article
author	José Manuel Borrero-de Acuña Cristian Hidalgo-Dumont Nicolás Pacheco Alex Cabrera Ignacio Poblete-Castro
author_facet	José Manuel Borrero-de Acuña Cristian Hidalgo-Dumont Nicolás Pacheco Alex Cabrera Ignacio Poblete-Castro
author_sort	José Manuel Borrero-de Acuña
title	A novel programmable lysozyme-based lysis system in Pseudomonas putida for biopolymer production
title_short	A novel programmable lysozyme-based lysis system in Pseudomonas putida for biopolymer production
title_full	A novel programmable lysozyme-based lysis system in Pseudomonas putida for biopolymer production
title_fullStr	A novel programmable lysozyme-based lysis system in Pseudomonas putida for biopolymer production
title_full_unstemmed	A novel programmable lysozyme-based lysis system in Pseudomonas putida for biopolymer production
title_sort	novel programmable lysozyme-based lysis system in pseudomonas putida for biopolymer production
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/1be251829cf2424999c089d62f752eda
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A novel programmable lysozyme-based lysis system in Pseudomonas putida for biopolymer production

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