Genetic engineering and screening of large number of cells or populations is a crucial bottleneck in today's systems biology and applied (micro)biology. Instead of using standard methods in bottles, flasks or 96-well plates, scientists are increasingly relying on high-throughput strategies that miniaturize their experiments to the nanoliter and picoliter scale and the single-cell level. In this review, we summarize different high-throughput system-wide genome engineering and screening strategies for microbes. More specifically, we will emphasize the use of multiplex automated genome evolution (MAGE) and CRISPR/Cas systems for high-throughput genome engineering and the application of (lab-on-chip) nanoreactors for high-throughput single-cell or population screening.

TZP is a BL/BLI recommended for the empirical treatment of severe infections. The excessive use of TZP has promoted the emergence of TZP-resistant Escherichia coli isolates.

HAGO CONSTAR que la presente memoria titulada "Nitrogen metabolism in wine yeast during alcoholic fermentation: effect on growth, fermentation activity and aroma production" presentada por la Srta. Alicia Gutiérrez Linares para optar al grado de Doctor en Ciencia, Tecnología y Gestión Alimentaria por la Universidad Politécnica de Valencia, ha sido realizada bajo nuestra dirección y supervisión, reuniendo las condiciones necesarias para ser defendida por su autora. Valencia, 20 de Junio de 2013Dr. José Manuel Guillamón Navarro Dra. Rosa Ana Chiva Tomás Instituto de Agroquímica y Tecnología de Alimentos (IATA-CSIC)Departamento de Biotecnología Grupo de Microbiología Molecular de Levaduras IndustrialesNitrogen metabolism in wine yeast during alcoholic fermentation: effect on growth, fermentation activity and aroma production ABSTRACT Nitrogen deficiency has been associated with major problems encountered in contemporary winemaking, especially those related to slow and incomplete fermentations. Under wine-making conditions, initial low levels of nitrogen act by limiting growth and biomass, resulting in a reduced fermentation rate. Nitrogen compounds present in the must have also influence on the production of volatile and non-volatile metabolites which regulate sensorial profile and quality of wine.Currently, the most common method for dealing with nitrogen-deficient fermentations is adding supplementary nitrogen. However, the effect of these additions is regulated by specific nitrogen requirements of each yeast strain and the conditions of fermentation.This doctoral thesis studies the individual nitrogen metabolism of four commercial wine strains, widely used in Spanish wine industry, especially regarding cell growth and fermentation activity, as well as metabolites production. This study has focused on the importance of nitrogen quantity and quality presented in the must, in order to achieve optimal fermentation performance. Through this work the effectiveness of different markers has been tested to find a biosensor that can be used to detect nitrogen limitation conditions under fermentation process. This work has been performed using biochemical, analytical and molecular approaches. Thus, some genetic variations involved in nitrogen utilization have been detected, allowing in the future the possibility to enhance fermentation performance of these strains.Metabolismo del nitrógeno en levaduras vínicas durante la fermentación alcohólica: efecto en el crecimiento, actividad fermentativa y producción de aromas RESUMENLa carencia de nitrógeno es uno de los principales problemas encontrados en la elaboración del vino, especialmente relacionados con fermentaciones lentas e incompletas. En condiciones de vinificación, bajos niveles iniciales de nitrógeno limitan el crecimiento y la producción de biomasa, provocando una baja tasa de fermentación. Los compuestos nitrogenados presentes en el mosto también influyen en la producción de metabolitos volátiles y no volátiles que son los que regulan el perfil sensorial y la calid...