title: Geranyl diphosphate synthase: an important regulation point in balancing a recombinant monoterpene pathway in Escherichia coli.
abstract: The expression level of geranyl diphosphate synthase (GPPS) was suspected to play a key role for geraniol production in recombinant Escherichia coli harboring an entire mevalonate pathway operon and a geraniol synthesis operon. The expression of GPPS was optimized by using ribosomal binding sites (RBSs) designed to have different translation initiation rates (TIRs). The RBS strength in TIR window of 500 arbitrary unit (au)-1400 au for GPPS appears to be suitable for balancing the geraniol biosynthesis pathway in this study. With the TIR of 500 au, the highest production titer of geraniol was obtained at a level of 1119mg/L, which represented a 6-fold increase in comparison with the previous titer of 183mg/L. The TIRs of GPPS locating out of range of the optimal window (500-1400 au) caused significant decreases of cell growth and geraniol production. It was suspected to result from metabolic imbalance and plasmid instability in geraniol production by inappropriate expression level of GPP synthase. Our results collectively indicated GPPS as an important regulation point in balancing a recombinant geraniol synthesis pathway. The GPPS-based regulation approach could be applicable for optimizing microbial production of other monoterpenes.
authors.affiliation: Division of Applied Life Science (BK21 Plus), PMBBRC, Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: Jia
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Division of Applied Life Science (BK21 Plus), PMBBRC, Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: Chonglong
authors.initials: C
authors.lastname: Wang
authors.affiliation: Division of Applied Life Science (BK21 Plus), PMBBRC, Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: Liyang
authors.initials: L
authors.lastname: Yang
authors.affiliation: Industrial Biotechnology Research Center, KRIBB, Daejeon 305-806, Republic of Korea. Electronic address: choi4162@kribb.re.kr.
authors.forename: Eui-Sung
authors.initials: ES
authors.lastname: Choi
authors.affiliation: Division of Applied Life Science (BK21 Plus), PMBBRC, Gyeongsang National University, Jinju 660-701, Republic of Korea. Electronic address: swkim@gnu.ac.kr.
authors.forename: Seon-Won
authors.initials: SW
authors.lastname: Kim
mesh_terms: D000080462:Acyclic Monoterpenes; D001665:Binding Sites / Q000235:genetics; D011756:Diphosphates; D004224:Diterpenes; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D029968:Escherichia coli Proteins; D015964:Gene Expression Regulation, Bacterial; D051228:Geranyltranstransferase; D008798:Mevalonic Acid / Q000378:metabolism; D039821:Monoterpenes / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D009876:Operon; D010442:Peptide Chain Initiation, Translational; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012270:Ribosomes / Q000378:metabolism; D013729:Terpenes / Q000378:metabolism
chemical_list: D000080462:Acyclic Monoterpenes; D011756:Diphosphates; D004224:Diterpenes; D029968:Escherichia coli Proteins; D039821:Monoterpenes; D011993:Recombinant Fusion Proteins; D013729:Terpenes; C511282:geranyl diphosphate; D051228:Geranyltranstransferase; C573354:IspA protein, E coli; C007836:geraniol; D008798:Mevalonic Acid
keywords: Escherichia coli; GPP synthesis; Geraniol; Mevalonate pathway; Plasmid maintenance; RBS strength