Publication: Desarrollo de herramientas moleculares para la producción de policétidos y péptidos no ribosomales
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2017-05-12
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Universidad Complutense de Madrid
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Abstract
Los ambientes marinos son una gran fuente de compuestos naturales. Muchas de estas moléculas son sintetizadas como mecanismo de defensa por microorganismos simbiontes de organismos marinos tales como esponjas, tunicados o poliquetos. Entre la gran variedad de productos naturales, cabe destacar la presencia de moléculas producidas por clústeres biosintéticos bacterianos que codifican policétido sintasas (PKS) y/o sintasas de péptidos no ribosomales (NRPS). La organización modular de este tipo de clústeres génicos, que en ocasiones pueden abarcar grandes regiones del genoma, permite la síntesis de compuestos con una gran variabilidad química. Existen diferencias en las herramientas utilizadas para identificar clústeres génicos que codifican PKSs y NRPSs, dependiendo de que se trate de un microorganismo cultivable o de un microbioma completo que incluye el microorganismo productor. En el caso de que se haya conseguido aislar y cultivar el microorganismo productor se pueden utilizar herramientas genómicas, como procedimientos de secuenciación masiva y su posterior análisis bioinformático, o bien la construcción de librerías genómicas en vectores de clonación de gran capacidad. Sin embargo, si se pretende identificar la secuencia de DNA de un clúster productor que pertenece a un microorganismo no cultivable, se pueden utilizar diferentes herramientas metagenómicas. Entre estas técnicas se encuentran la generación de librerías metagenómicas a partir de DNA metagenómico obtenido del microbioma y procesos de secuenciación masiva para su posterior análisis mediante herramientas bioinformáticas especializadas...
Marine environments are a huge source of natural compounds. Many of these molecules are synthesized as a defense mechanism by symbiotic microorganisms found in marine organisms such as sponges, tunicates and polychaetes. Among the wide diversity of natural products, the presence of molecules produced by bacterial biosynthetic clusters that encode polyketide synthases (PKS) and/or non-ribosomal peptide synthases (NRPS) is remarkable. The modular organization of these gene clusters, which sometimes cover large genome regions, allows the synthesis of compounds that can present a huge chemical variability. Depending on whether it concerns a culturable microorganism or a full microbiome, there are some differences in the tools used to identify PKSs and/or NPRSs gene clusters. If the producer microorganism has been isolated and cultured, some genomic tools such as massive sequencing procedures (including subsequent bioinformatics analysis) or genomic libraries cloned in high capacity vectors can be used. However, if the DNA sequence of interest belongs to a non-culturable microorganism, different metagenomics tools can be used. Among these techniques it is possible to find the generation of metagenomic libraries from environmental DNA and processes using massive sequencing that include analysis performed with specialized bioinformatics tools...
Marine environments are a huge source of natural compounds. Many of these molecules are synthesized as a defense mechanism by symbiotic microorganisms found in marine organisms such as sponges, tunicates and polychaetes. Among the wide diversity of natural products, the presence of molecules produced by bacterial biosynthetic clusters that encode polyketide synthases (PKS) and/or non-ribosomal peptide synthases (NRPS) is remarkable. The modular organization of these gene clusters, which sometimes cover large genome regions, allows the synthesis of compounds that can present a huge chemical variability. Depending on whether it concerns a culturable microorganism or a full microbiome, there are some differences in the tools used to identify PKSs and/or NPRSs gene clusters. If the producer microorganism has been isolated and cultured, some genomic tools such as massive sequencing procedures (including subsequent bioinformatics analysis) or genomic libraries cloned in high capacity vectors can be used. However, if the DNA sequence of interest belongs to a non-culturable microorganism, different metagenomics tools can be used. Among these techniques it is possible to find the generation of metagenomic libraries from environmental DNA and processes using massive sequencing that include analysis performed with specialized bioinformatics tools...
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Tesis inédita de la Universidad Complutense de Madrid, Facultad de Ciencias Químicas, Departamento de Bioquímica y Biología Molecular, leída el 17-12-2015