Profiling proteomic responses to hexokinase-II depletion in terpene-producing Saccharomyces cerevisiae  

作　　者：Zeyu Lu Qianyi Shen Lian Liu Gert Talbo Robert Speight Matt Trau Geoff Dumsday Christopher B.Howard Claudia E.Vickers Bingyin Peng 

机构地区：[1]ARC Centre of Excellence in Synthetic Biology,Queensland University of Technology,Brisbane,QLD,4000,Australia [2]Australian Institute for Bioengineering and Nanotechnology(AIBN),The University of Queensland,Brisbane,QLD,4072,Australia [3]Centre of Agriculture and the Bioeconomy,School of Biology and Environmental Science,Faculty of Science,Queensland University of Technology,Brisbane,QLD,4000,Australia [4]CSIRO Synthetic Biology Future Science Platform,Commonwealth Scientific and Industrial Research Organisation(CSIRO),Black Mountain,ACT,2601,Australia [5]School of Chemistry and Molecular Biosciences(SCMB),the University of Queensland,Brisbane,QLD,4072,Australia [6]Metabolomics Australia(Queensland Node),Australian Institute for Bioengineering and Nanotechnology(AIBN),The University of Queensland,Brisbane,QLD,4072,Australia [7]CSIRO Manufacturing,Clayton,VIC,3169,Australia [8]Eden Brew Pty Ltd,Glenorie,NSW,2157,Australia

出　　处：《Engineering Microbiology》2023年第3期1-13,共13页工程微生物学（英文）

基　　金：supported by Australian Research Council centre of Excellence in Synthetic Biology(CE200100029);supported by BioPlatforms Australia through the Commonwealth Government’s National Collaborative Research Infrastructure Strategy(NCRIS).

摘　　要：Hexokinase II(Hxk2)is a master protein in glucose-mediated transcriptional repression signaling pathway.De-grading Hxk2 through an auxin-inducible protein degradation previously doubled sesquiterpene(nerolidol)pro-duction at gram-per-liter levels in Saccharomyces cerevisiae.Global transcriptomics/proteomics profiles in Hxk2-deficient background are important to understanding genetic and molecular mechanisms for improved nerolidol production and guiding further strain optimization.Here,proteomic responses to Hxk2 depletion are investi-gated in the yeast strains harboring a GAL promoters-controlled nerolidol synthetic pathway,at the exponential and ethanol growth phases and in GAL80-wildtype and gal80Δbackgrounds.Carbon metabolic pathways and amino acid metabolic pathways show diversified responses to Hxk2 depletion and growth on ethanol,including upregulation of alternative carbon catabolism and respiration as well as downregulation of amino acid synthesis.De-repression of GAL genes may contribute to improved nerolidol production in Hxk2-depleted strains.Seven-teen transcription factors associated with upregulated genes are enriched.Validating Ash1-mediated repression on the RIM4 promoter shows the variation on the regulatory effects of different Ash1-binding sites and the syner-gistic effect of Ash1 and Hxk2-mediated repression.Further validation of individual promoters shows that HXT1 promoter activities are glucose-dependent in hxk2Δbackground,but much weaker than those in HXK2-wildtype background.In summary,inactivating HXK2 may relieve glucose repression on respiration and GAL promoters for improved bioproduction under aerobic conditions in S.cerevisiae.The proteomics profiles provide a better genetics overview for a better metabolic engineering design in Hxk2-deficient backgrounds.

关 键 词：PROTEOMICS Glucose repression Metabolic engineering Crabtree effect SESQUITERPENE Synthetic biology Genetic regulation 

分 类 号：Q93[生物学—微生物学]