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

作者：Borrero de Acuna Jose Manuel; Hidalgo Dumont Cristian; Pacheco Nicolas; Cabrera Alex; Poblete Castro Ignacio

来源：Scientific Reports, 2017, 7(1): 4373.

DOI：10.1038/s41598-017-04741-2

<jats:title>Abstract</jats:title><jats:p>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 <jats:italic>Escherichia coli</jats:italic> and <jats:italic>Pseudomonas putida</jats:italic> KT2440. Before induction, the lytic system did not significantly arrest essential physiological parameters in the recombinant <jats:italic>E</jats:italic>. <jats:italic>coli</jats:italic> (ECPi) and <jats:italic>P</jats:italic>. <jats:italic>putida</jats:italic> (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.</jats:p>

• 出版日期2017-6-29