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Implementation of an autolysis systems as a tool for functional metagenomics analysis

AuthorsCárcel Márquez, J.; Camacho, Eva María ; Flores, Amando ; Santero, Eduardo
Issue Date2017
CitationFEMS 2017
Abstract[Backgrounds]: Functional metagenomic allows for the identification of novel activities of interest present in uncultured microorganisms. Unfortunately, this strategy is limited, among other factors, by the accessibility of the heterologous enzyme to the substrate. For intracellular activities, the substrate must enter the bacterial cell to allow detection, while for extracellular activities the host must be able to secrete the enzyme to the growth medium. This way the efficiency of the screening could be limited. [Objectives]: The main purpose of this work is to develop an autolysis system that, in response to an inducer, lysates bacteria releasing their content into the growth medium. This system must also permit survival of a proportion of the bacterial population to allow the recovery of positive clones. [Methods]: We have implemented an inducible autolysis system previously developed in our laboratory. It is based in the lysis operon of lambda phage and responds to anhydrotetracycline as an inducer. We have demonstrated that the autolysis system allows detection of intracellular activities and the recovery of positive clones. Subsequently we carried out a screening for cellullase activity in the presence or absence of the autolysis system to compare the efficiency of cellulose activity detection. [Conclusions]: We have demonstrated that the autolysis system is necessary for the detection of intracellular activities. Additionally, the system improves detection of positive clones for cellullase activity. In all cases, the lysis system allows recovery and subsequent confirmation of positive clones. In summary, our system is an effective tool to improve functional metagenomic analysis.
DescriptionResumen del trabajo presentado al 7th Congress of European Microbiologists, celebrado en Valencia (España) del 9 al 13 de julio de 2017.
Appears in Collections:(CABD) Comunicaciones congresos
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