4.2.3.19 A676T site-directed mutagenesis of isozyme RcKSL2, mutation of the catalytically important residue results in 60fold increased product amount composed of essentially the same mix of ent-trachylobane and ent-kaurene as observed for the wild-type enzyme 748876 4.2.3.19 A710C no change in reaction product profile compared to wild-type 714971 4.2.3.19 A710F mutant converts geranylgeranyl diphosphate to ent-kaurene as the sole product 714971 4.2.3.19 A710G no change in reaction product profile compared to wild-type 714971 4.2.3.19 A710M mutant converted geranylgeranyl diphosphate to ent-kaurene as the sole product 714971 4.2.3.19 A710N no change in reaction product profile compared to wild-type 714971 4.2.3.19 A710S no change in reaction product profile compared to wild-type 714971 4.2.3.19 D132A low activity 492252 4.2.3.19 D320A low activity 492252 4.2.3.19 D635A/D636A mutation of the DDYFD motif in the kaurene synthase domain 679843 4.2.3.19 D656A no activity 492252 4.2.3.19 D75A site-directed mutagenesis -, 749301 4.2.3.19 D75C site-directed mutagenesis, mutant crystal structure determination and analysis -, 749301 4.2.3.19 D79C site-directed mutagenesis -, 749301 4.2.3.19 H212A the mutant shows altered substrate specificity compared to wild-type, the single histidine to alanine mutation converts FfCPS/KS into a bifunctional ent-13-epi-manoyl oxide synthase -, 747507 4.2.3.19 H302A the mutant efficiently produces ent-13-epi-manoyl oxide from geranylgeranyl diphosphate 747507 4.2.3.19 H302A the mutant shows altered substrate specificity compared to wild-type, the single histidine to alanine mutation converts PpCPS/KS into a bifunctional ent-13-epi-manoyl oxide synthase 747507 4.2.3.19 I368A the mutant produces ent-pimara-8(14),15-diene, ent-kaur-16-ene, 8alpha-hydroxy-ent-pimara-15-ene, and ent-pimara-7,15-diene from ent-copalyl diphosphate 747870 4.2.3.19 I368S the mutant produces ent-pimara-8(14),15-diene and 8alpha-hydroxy-ent-pimara-15-ene from ent-copalyl diphosphate 747870 4.2.3.19 I368T the mutant produces ent-pimara-8(14),15-diene from ent-copalyl diphosphate 747870 4.2.3.19 I368V the mutant produces ent-kaur-16-ene from ent-copalyl diphosphate 747870 4.2.3.19 I619A site-directed mutagenesis, the mutant shows altered product spectrum compared to the wild-type enzyme 747870 4.2.3.19 I638A site-directed mutagenesis, the mutant shows altered product spectrum compared to the wild-type enzyme, the mutant produces a mixture of four products, pedominantly ent-pimara-8(14),15-diene, and a small amount of the double bond isomer ent-pimara-7,15-diene, and variable amounts of ent-kaur-16-ene, as well as substantial amounts of 8a-hydroxy-ent-pimar-15-ene 747870 4.2.3.19 I638S site-directed mutagenesis, the mutant shows altered product spectrum compared to the wild-type enzyme, the mutant produces a mixture of four products, pedominantly ent-pimara-8(14),15-diene, and a small amount of the double bond isomer ent-pimara-7,15-diene, and variable amounts of ent-kaur-16-ene, as well as substantial amounts of 8a-hydroxy-ent-pimar-15-ene 747870 4.2.3.19 I638T site-directed mutagenesis, the mutant shows altered product spectrum compared to the wild-type enzyme, the mutant produces a mixture of four products, pedominantly ent-pimara-8(14),15-diene, and small amounts of the double bond isomer ent-pimara-7,15-diene and 8a-hydroxy-ent-pimar-15-ene, and variable amounts of ent-kaur-16-ene 747870 4.2.3.19 I638V site-directed mutagenesis, the mutant shows unaltered product spectrum compared to the wild-type enzyme and produces just ent-kaur-16-ene 747870 4.2.3.19 I645T site-directed mutagenesis, the mutant shows altered product spectrum compared to the wild-type enzyme, the mutant produces almost entirely ent-pimara-8(14),15-diene, reaction of EC 4.2.3.30 747870 4.2.3.19 I645T the mutant produces ent-pimara-8(14),15-diene from ent-copalyl diphosphate 747870 4.2.3.19 I664A site-directed mutagenesis, the mutant shows altered product spectrum compared to the wild-type enzyme, the mutant produces significant amounts of 8alpha-hydroxy-ent-pimara-15-ene generated by addition of water prior to deprotonation, it also produces ent-copalol, the dephosphorylated derivative of ent-copalyl diphosphate 747870 4.2.3.19 I664S site-directed mutagenesis, the mutant shows altered product spectrum compared to the wild-type enzyme, the mutant produces significant amounts of 8alpha-hydroxy-ent-pimara-15-ene generated by addition of water prior to deprotonation, it also produces ent-copalol, the dephosphorylated derivative of ent-copalyl diphosphate 747870 4.2.3.19 I664T site-directed mutagenesis, the mutant shows altered product spectrum compared to the wild-type enzyme, the mutant produces high amounts of ent-pimara-8(14),15-diene, it also produces ent-copalol, the dephosphorylated derivative of ent-copalyl diphosphate 747870 4.2.3.19 I664V site-directed mutagenesis, the mutant shows altered product spectrum compared to the wild-type enzyme, the mutant produces ent-isokaurene and ent-copalol, the dephosphorylated derivative of ent-copalyl diphosphate 747870 4.2.3.19 I741T site-directed mutagenesis, the mutant shows altered product spectrum compared to the wild-type enzyme, the mutant produces almost entirely ent-pimara-8(14),15-diene, reaction of EC 4.2.3.30 747870 4.2.3.19 I741T the mutant produces ent-pimara-8(14),15-diene from ent-copalyl diphosphate 747870 4.2.3.19 additional information a synthetic ent-kaurene module, expressing Stevia rebaudiana CPPS and KS, and Rhodobacter sphaeroides GGPPS (crtE gene), is constructed for ent-kaurene production in diverse Escherichia coli strains, best in strain MG1655, method optimization, several crtE genes from Rhodobacter species are validated, overview 748267 4.2.3.19 additional information construction of a SmKS-SmCPSent fusion enzyme (CPSent is ent-copalyl diphosphate synthase, EC 5.5.1.13), which improves the ent-kaurene precursor supply in transformed Saccharomyces cerevisiae strain SGH5 (BY-T20/pESC-Trp::SmKS-SmCPSent/SmKO + pESC-Leu::SmCPR1) 746448 4.2.3.19 additional information construction of chimeric proteins of Physcomitrella patens PpCPS/KS with Jungermannia subulata, the chimeric cyclases Pp131-566 /Js574-86 and Pp131-504 /Js512-886 and produce only ent-kaurene. Chimeric cyclases, Pp131-622 /Js630-886 and Pp131-714 /Js722-886, lose all enzymic activity 714971 4.2.3.19 additional information construction of chimeric proteins of Physcomitrella patens PpCPS/KS with Jungermannia subulata, the chimeric cyclases Pp131-566 /Js574-886 and Pp131-504 /Js512-886 and produce only ent-kaurene. Chimeric cyclases, Pp131-622 /Js630-886 and Pp131-714 /Js722-886, lose all enzymic activity 714971 4.2.3.19 additional information construction of transgenic Arabidopsis thaliana plants, using the Agrobacterium tumefaciens infection system, overexpressing KS leads to increased ent-kaurene production but not to an increase in bioactive gibberellins, no altered morphology or phenotype compared to wild-type plants 666619 4.2.3.19 additional information construction of transgenic rice mutants expressing mutated KS-like genes, the mutant plants show reduced enzyme activity, phenotype analysis and effects on gibberellin metabolism, overview 660237 4.2.3.19 additional information generation of a Physcomitrella patens CPS/KS (PpCPS/KS)-gene disruption mutant strain A74, comparisons of photomorphogenesis of protonema in the wild-type and the A74 enzyme mutant, phenotype, detailed overview 749028 4.2.3.19 additional information several N- and C-terminal truncated enzymes produced, all found to be inactive 492252 4.2.3.19 R204A site-directed mutagenesis -, 749301