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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| Format: | Article |
| Language: | English |
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Nature Portfolio
2017-06-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-017-04741-2 |
| _version_ | 1818422033394958336 |
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| 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 |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-14T13:19:49Z |
| format | Article |
| id | doaj.art-1be251829cf2424999c089d62f752eda |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-14T13:19:49Z |
| publishDate | 2017-06-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-1be251829cf2424999c089d62f752eda2022-12-21T22:59:57ZengNature PortfolioScientific Reports2045-23222017-06-017111110.1038/s41598-017-04741-2A novel programmable lysozyme-based lysis system in Pseudomonas putida for biopolymer productionJosé Manuel Borrero-de Acuña0Cristian Hidalgo-Dumont1Nicolás Pacheco2Alex Cabrera3Ignacio Poblete-Castro4Biosystems Engineering Laboratory, Center for Bioinformatics and Integrative Biology (CBIB), Faculty of Biological Sciences, Universidad Andres BelloBiosystems Engineering Laboratory, Center for Bioinformatics and Integrative Biology (CBIB), Faculty of Biological Sciences, Universidad Andres BelloBiosystems Engineering Laboratory, Center for Bioinformatics and Integrative Biology (CBIB), Faculty of Biological Sciences, Universidad Andres BelloBiosystems Engineering Laboratory, Center for Bioinformatics and Integrative Biology (CBIB), Faculty of Biological Sciences, Universidad Andres BelloBiosystems Engineering Laboratory, Center for Bioinformatics and Integrative Biology (CBIB), Faculty of Biological Sciences, Universidad Andres BelloAbstract 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.https://doi.org/10.1038/s41598-017-04741-2 |
| spellingShingle | 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 Scientific Reports |
| title | 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_short | 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 |
| url | https://doi.org/10.1038/s41598-017-04741-2 |
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