Bioethanol production from lignocellulosicbiomass: assessing genes linked to acetic acid stress in Saccharomyces cerevisiae Public

http://ir.library.oregonstate.edu/concern/honors_college_theses/qn59q584g

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  • Lignocellulosic biomass represents a vast and renewable source of fermentable sugar for production of biofuels. However, native lignocellulose—comprised of cellulose, hemicellulose and lignin—is refractory to degradation because the crystalline cellulose is not easily hydrolyzed by cellulases. Standard chemical treatments of lignocellulose to reduce the crystallinity of cellulose prior to enzymatic hydrolysis also generate fermentation inhibitors, including acetic and other organic acids, furfural, hydroxymethyl furfural, and phenolic compounds. To increase the acetic acid resistance of one of the major microorganisms used to ferment lignocellulose-derived sugars, mutations in three genes were introduced and evaluated in a prototrophic laboratory strain of Saccharomyces cerevisiae, S288c, and were introduced into the industrial strain D5A. Previous work had shown that loss of FPS1, encoding an acetic acid channel, and loss of EDE1 and MVB12, encoding proteins involved in endocytosis, increased acetic acid resistance. Disruption of FPS1 in S288c resulted in somewhat greater resistance at low concentrations (80-120 mM acetic acid) but not at concentrations greater than 120 mM. Combined genetic and molecular analyses of the disruptions of FPS1, EDE1 and MVB12 in D5A indicated that D5A was diploid, and that only one of the two alleles of each of these genes had been deleted.
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