Quantitative Metabolomics and Its Application in Metabolic Engineering of Microbial Cell Factories Exemplified by the Baker’s Yeast

Abstract

Metabolomics 20 interesting precursors for pharmaceuticals (Csuk 1991). It can grow as a diploid as well as a haploid which highly facilitates genetic manipulation and permits high-throughput genetic engineering. Considering the enormous early interest in studying and understanding the physiology of S. cerevisiae long before modern omics techniques have been developed, it is not very surprising that it was the baker's yeasts genome that was the first within the domain of eukaryotes that was completely sequenced. Genomic and biological information about S. cerevisiae molecular biology is comprehensively collected at the Saccharomyces Genome Database (SGD, http://www.yeastgenome.org/). Driven by the knowledge of the complete genomic sequence and by the steadily increasing availability of tools developed for genetic engineering, S. cerevisiae became a key work horse and the representative eukaryotic model organism in every modern discipline within the biosciences such as molecular and cell biology, functional genomics, systems biology or metabolic and synthetic engineering. Today's genetic work with S. cerevisiae cells is highly alleviated by the presence of a wide spectrum of established yeast molecular biology tool kits and availability of many wild-type and mutant strain (e.g. knockout strains) collections as well as plasmid collections containing S. cerevisiae ORFs, gene deletion markers or promoter sets and many more, offered by commercial sources such as EUROSCARF

Targeting the H3K4 Demethylase KDM5B Reprograms the Metabolome and Phenotype of Melanoma Cells

“…A total of 75,000 cells were seeded onto lumox membranes (Sarstedt AG & Co). Medium was changed after 24 hours, and 48 hours after changing medium, cells and supernatants were harvested in 6 dish batches as described and quenched in boiling ethanol (Klimacek, 2012). From the supernatant, 500 ml was taken and centrifuged for 1 minute at 14,000g and 4 C. An aliquot of 100 ml was transferred into 2 ml 80%/20% ethanol/ammonium acetate (85 C-preheated).…”