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      Transcriptomic Analyses during the Transition from Biomass Production to Lipid Accumulation in the Oleaginous Yeast Yarrowia lipolytica

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          Abstract

          We previously developed a fermentation protocol for lipid accumulation in the oleaginous yeast Y. lipolytica. This process was used to perform transcriptomic time-course analyses to explore gene expression in Y. lipolytica during the transition from biomass production to lipid accumulation. In this experiment, a biomass concentration of 54.6 g CDW/l, with 0.18 g/g CDW lipid was obtained in ca. 32 h, with low citric acid production. A transcriptomic profiling was performed on 11 samples throughout the fermentation. Through statistical analyses, 569 genes were highlighted as differentially expressed at one point during the time course of the experiment. These genes were classified into 9 clusters, according to their expression profiles. The combination of macroscopic and transcriptomic profiles highlighted 4 major steps in the culture: (i) a growth phase, (ii) a transition phase, (iii) an early lipid accumulation phase, characterized by an increase in nitrogen metabolism, together with strong repression of protein production and activity; (iv) a late lipid accumulation phase, characterized by the rerouting of carbon fluxes within cells. This study explores the potential of Y. lipolytica as an alternative oil producer, by identifying, at the transcriptomic level, the genes potentially involved in the metabolism of oleaginous species.

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          Gene Ontology: tool for the unification of biology

          Genomic sequencing has made it clear that a large fraction of the genes specifying the core biological functions are shared by all eukaryotes. Knowledge of the biological role of such shared proteins in one organism can often be transferred to other organisms. The goal of the Gene Ontology Consortium is to produce a dynamic, controlled vocabulary that can be applied to all eukaryotes even as knowledge of gene and protein roles in cells is accumulating and changing. To this end, three independent ontologies accessible on the World-Wide Web (http://www.geneontology.org) are being constructed: biological process, molecular function and cellular component.
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            Normalization of cDNA microarray data.

            Normalization means to adjust microarray data for effects which arise from variation in the technology rather than from biological differences between the RNA samples or between the printed probes. This paper describes normalization methods based on the fact that dye balance typically varies with spot intensity and with spatial position on the array. Print-tip loess normalization provides a well-tested general purpose normalization method which has given good results on a wide range of arrays. The method may be refined by using quality weights for individual spots. The method is best combined with diagnostic plots of the data which display the spatial and intensity trends. When diagnostic plots show that biases still remain in the data after normalization, further normalization steps such as plate-order normalization or scale-normalization between the arrays may be undertaken. Composite normalization may be used when control spots are available which are known to be not differentially expressed. Variations on loess normalization include global loess normalization and two-dimensional normalization. Detailed commands are given to implement the normalization techniques using freely available software.
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              Microbial production of fatty-acid-derived fuels and chemicals from plant biomass.

              Increasing energy costs and environmental concerns have emphasized the need to produce sustainable renewable fuels and chemicals. Major efforts to this end are focused on the microbial production of high-energy fuels by cost-effective 'consolidated bioprocesses'. Fatty acids are composed of long alkyl chains and represent nature's 'petroleum', being a primary metabolite used by cells for both chemical and energy storage functions. These energy-rich molecules are today isolated from plant and animal oils for a diverse set of products ranging from fuels to oleochemicals. A more scalable, controllable and economic route to this important class of chemicals would be through the microbial conversion of renewable feedstocks, such as biomass-derived carbohydrates. Here we demonstrate the engineering of Escherichia coli to produce structurally tailored fatty esters (biodiesel), fatty alcohols, and waxes directly from simple sugars. Furthermore, we show engineering of the biodiesel-producing cells to express hemicellulases, a step towards producing these compounds directly from hemicellulose, a major component of plant-derived biomass.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2011
                22 November 2011
                : 6
                : 11
                : e27966
                Affiliations
                [1 ]INRA, UMR1319 Micalis, Jouy-en-Josas, France
                [2 ]Université de Toulouse, INSA, UPS, INP, LISBP, Toulouse, France
                [3 ]INRA, UMR792, Ingénierie des Systèmes Biologiques et des Procédés, Toulouse, France
                [4 ]CNRS, UMR5504, Toulouse, France
                [5 ]Dynamique des Structures et Interactions des Macromolécules Biologiques, UMR-S 665 - Université Paris 7, INTS, Paris, France
                [6 ]Plateforme Biopuces de la Génopole de Toulouse Midi Pyrénées, INSA/DGBA 135, Toulouse, France
                [7 ]CNRS, Micalis, Jouy-en-Josas, France
                Texas A&M University, United States of America
                Author notes

                Conceived and designed the experiments: JMN JC CMJ JLU. Performed the experiments: JC VLB. Analyzed the data: NM JC GL JMN AB. Contributed reagents/materials/analysis tools: VLB CMJ JMN. Wrote the paper: NM JC AB JMN.

                Article
                PONE-D-11-10642
                10.1371/journal.pone.0027966
                3222671
                22132183
                e3b90380-c6bb-486d-abd7-0259e08e3643
                Morin et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
                History
                : 21 June 2011
                : 28 October 2011
                Page count
                Pages: 13
                Categories
                Research Article
                Biology
                Biochemistry
                Lipids
                Lipid Metabolism
                Metabolism
                Lipid Metabolism
                Nitrogen Metabolism
                Proteins
                Lipoproteins
                Protein Interactions
                Regulatory Proteins
                Genetics
                Gene Expression
                Protein Translation
                Microbiology
                Mycology
                Yeast
                Model Organisms
                Yeast and Fungal Models

                Uncategorized
                Uncategorized

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