1,11-ring closure of (2E,6E)-farnsyl diphosphate yields a highly reactive trans-humulyl carbocation. A 1,2-hydride shift followed by further ring closures results in the protoilludyl cation, which undergoes a final deprotonation to give DELTA6-protoilludene
in the presence of Ca2+, DELTA6-protoilludene synthases catalyze diverse cyclization reactions. DELTA6-Protoilludene and (E)-beta-caryophyllene result from a 1,11 cyclization of (2E,6E)-farnesyl diphosphate following metal ion mediated ionization. The intermediate trans-humulyl cation may undergo a hydride shift followed by two cyclizations and a deprotonation to yield DELTA6-protoilludene. Proposed mechanisms for sesquiterpene production by DELTA6-protoilludene synthases, overview
in the presence of Ca2+, DELTA6-protoilludene synthases catalyze diverse cyclization reactions. DELTA6-Protoilludene and (E)-beta-caryophyllene result from a 1,11 cyclization of (2E,6E)-FPP following metal ion mediated ionization. The intermediate trans-humulyl cation may undergo a hydride shift followed by two cyclizations and a deprotonation to yield DELTA6-protoilludene. Proposed mechanisms for sesquiterpene production by DELTA6-protoilludene synthases, overview
in the presence of Ca2+, DELTA6-protoilludene synthases catalyze diverse cyclization reactions. DELTA6-Protoilludene and (E)-beta-caryophyllene result from a 1,11-cyclization of (2E,6E)-farnesyl diphosphate following metal ion mediated ionization. The intermediate trans-humulyl cation may undergo a hydride shift followed by two cyclizations and a deprotonation to yield DELTA6-protoilludene. Proposed mechanisms for sesquiterpene production by DELTA6-protoilludene synthases, overview
main product for isoforms 25180, 64702, 73029. Isoforms 64702 and 73029 appear to be less specific than other previously described DELTA6-protoilludene synthases, and are capable of simultaneously producing a 1,10-cyclization-derived scaffold, beta-elemene, when expressed in a heterologous host
in the presence of Ca2+, DELTA6-protoilludene synthases catalyze diverse cyclization reactions. DELTA6-Protoilludene and (E)-beta-caryophyllene result from a 1,11 cyclization of (2E,6E)-FPP following metal ion mediated ionization. The intermediate trans-humulyl cation may undergo a hydride shift followed by two cyclizations and a deprotonation to yield DELTA6-protoilludene. Diphosphate enzyme-coupled spectrophotometric assay
in the presence of Ca2+, DELTA6-protoilludene synthases catalyze diverse cyclization reactions. DELTA6-Protoilludene and (E)-beta-caryophyllene result from a 1,11 cyclization of (2E,6E)-farnesyl diphosphate following metal ion mediated ionization. The intermediate trans-humulyl cation may undergo a hydride shift followed by two cyclizations and a deprotonation to yield DELTA6-protoilludene. Diphosphate enzyme-coupled spectrophotometric assay
in the presence of Ca2+, DELTA6-protoilludene synthases catalyze diverse cyclization reactions. DELTA6-Protoilludene and (E)-beta-caryophyllene result from a 1,11-cyclization of (2E,6E)-farnesyl diphosphate following metal ion mediated ionization. The intermediate trans-humulyl cation may undergo a hydride shift followed by two cyclizations and a deprotonation to yield DELTA6-protoilludene Diphosphate enzyme-coupled spectrophotometric assay
DELTA6-protoilludene synthases are catalytically functional with a range of metal ions. Sesquiterpene synthases typically bind the divalent metal cations Mg2+ and Mn2+ to ionize the pyrophosphate of (2E,6E)-farnesyl diphosphate, exploration of metal ion dependencies of different DELTA6-protoilludene synthases, overview. Metal-dependent ionization results in the release of the diphosphate moiety from farnesyl diphosphate ((2E,6E)-FPP) leading to either a 1,11- or 1,10-cyclization of the primary carbocation. A 1,11-cyclization leads to the trans-humulyl cation, a 1,10-cyclization of the primary carbocation leads to the (E,E)-germacradienyl cation. Further hydride shifts and cyclizations results in final sesquiterpene products
DELTA6-protoilludene synthases are catalytically functional with a range of metal ions. Sesquiterpene synthases typically bind the divalent metal cations Mg2+ and Mn2+ to ionize the pyrophosphate of (2E,6E)-farnesyl diphosphate, exploration of metal ion dependencies of different DELTA6-protoilludene synthases, overview. Metal-dependent ionization results in the release of the diphosphate moiety from farnesyl diphosphate ((2E,6E)-FPP) leading to either a 1,11- or 1,10-cyclization of the primary carbocation. A 1,11 cyclization leads to the trans-humulyl cation, a 1,10-cyclization of the primary carbocation leads to the (E,E)-germacradienyl cation. Further hydride shifts and cyclizations results in final sesquiterpene products
DELTA6-protoilludene synthases are catalytically functional with a range of metal ions. Sesquiterpene synthases typically bind the divalent metal cations Mg2+ and Mn2+ to ionize the pyrophosphate of (2E,6E)-farnesyl diphosphate, exploration of metal ion dependencies of different DELTA6-protoilludene synthases, overview. Metal-dependent ionization results in the release of the diphosphate moiety from farnesyl diphosphate ((2E,6E)-FPP) leading to either a 1,11- or 1,10-cyclization of the primary carbocation. A 1,11-cyclization leads to the trans-humulyl cation, a 1,10-cyclization of the primary carbocation leads to the (E,E)-germacradienyl cation. Further hydride shifts and cyclizations results in final sesquiterpene products
in the presence of Ca2+, DELTA6-protoilludene synthases catalyze diverse cyclization reactions. DELTA6-Protoilludene and (E)-beta-caryophyllene result from a 1,11 cyclization of (2E,6E)-farnesyl diphosphate ((2E,6E)-FPP) following metal ion mediated ionization. The intermediate trans-humulyl cation may undergo a hydride shift followed by two cyclizations and a deprotonation to yield DELTA6-protoilludene. Alternatively, a direct 2,10 cyclization of the trans-humulyl cation leads to (E)-beta-caryophyllene. On the other hand, beta-elemene, 4,11-selinadiene, beta-selinene and alpha-selinene are derived from a 1,10 cyclization of (2E,6E)-FPP. A direct deprotonation of the intermediate (E,E)-germacradienyl cation results in germacrene A. beta-Elemene is the heat-induced Cope rearrangement product. A 2,7 cyclization of the (E,E)-germacradienyl cation is followed by deprotonation yields 4,11-selinadiene. Finally, a hydride shift, followed by a 2,7 cyclization of the (E,E)-germacradienyl cation and deprotonation leads to beta-selinene and alpha-selinene. Therefore DELTA6-protoilludene synthases are capable of catalyzing simultaneously a 1,10 and a 1,11 cyclization in the presence of an alternative metal ion
in the presence of Ca2+, DELTA6-protoilludene synthases catalyze diverse cyclization reactions. DELTA6-Protoilludene and (E)-beta-caryophyllene result from a 1,11 cyclization of (2E,6E)-farnesyl diphosphate ((2E,6E)-FPP) following metal ion mediated ionization. The intermediate trans-humulyl cation may undergo a hydride shift followed by two cyclizations and a deprotonation to yield DELTA6-protoilludene. Alternatively, a direct 2,10 cyclization of the trans-humulyl cation leads to (E)-beta-caryophyllene. On the other hand, beta-elemene, 4,11-selinadiene, beta-selinene and alpha-selinene are derived from a 1,10 cyclization of (2E,6E)-FPP. A direct deprotonation of the intermediate (E,E)-germacradienyl cation results in germacrene A. beta-Elemene is the heat-induced Cope rearrangement product. A 2,7 cyclization of the (E,E)-germacradienyl cation is followed by deprotonation yields 4,11-selinadiene. Finally, a hydride shift, followed by a 2,7 cyclization of the (E,E)-germacradienyl cation and deprotonation leads to beta-selinene and alpha-selinene. Therefore DELTA6-protoilludene synthases are capable of catalyzing simultaneously a 1,10 and a 1,11 cyclization in the presence of an alternative metal ion
in the presence of Ca2+, DELTA6-protoilludene synthases catalyze diverse cyclization reactions. DELTA6-Protoilludene and (E)-beta-caryophyllene result from a 1,11 cyclization of (2E,6E)-farnesyl diphosphate ((2E,6E)-FPP) following metal ion mediated ionization. The intermediate trans-humulyl cation may undergo a hydride shift followed by two cyclizations and a deprotonation to yield DELTA6-protoilludene. Alternatively, a direct 2,10 cyclization of the trans-humulyl cation leads to (E)-beta-caryophyllene. On the other hand, beta-elemene, 4,11-selinadiene, beta-selinene and alpha-selinene are derived from a 1,10 cyclization of (2E,6E)-FPP. A direct deprotonation of the intermediate (E,E)-germacradienyl cation results in germacrene A. beta-Elemene is the heat-induced Cope rearrangement product. A 2,7 cyclization of the (E,E)-germacradienyl cation is followed by deprotonation yields 4,11-selinadiene. Finally, a hydride shift, followed by a 2,7 cyclization of the (E,E)-germacradienyl cation and deprotonation leads to beta-selinene and alpha-selinene. Therefore DELTA6-protoilludene synthases are capable of catalyzing simultaneously a 1,10- and a 1,11-cyclization in the presence of an alternative metal ion
in the presence of Ca2+, DELTA6-protoilludene synthases catalyze diverse cyclization reactions. DELTA6-Protoilludene and (E)-beta-caryophyllene result from a 1,11 cyclization of (2E,6E)-farnesyl diphosphate ((2E,6E)-FPP) following metal ion mediated ionization. The intermediate trans-humulyl cation may undergo a hydride shift followed by two cyclizations and a deprotonation to yield DELTA6-protoilludene. Alternatively, a direct 2,10-cyclization of the trans-humulyl cation leads to (E)-beta-caryophyllene. On the other hand, beta-elemene, 4,11-selinadiene, beta-selinene and alpha-selinene are derived from a 1,10 cyclization of (2E,6E)-FPP. A direct deprotonation of the intermediate (E,E)-germacradienyl cation results in germacrene A. beta-Elemene is the heat-induced Cope rearrangement product. A 2,7 cyclization of the (E,E)-germacradienyl cation is followed by deprotonation yields 4,11-selinadiene. Finally, a hydride shift, followed by a 2,7 cyclization of the (E,E)-germacradienyl cation and deprotonation leads to beta-selinene and alpha-selinene. Therefore DELTA6-protoilludene synthases are capable of catalyzing simultaneously a 1,10- and a 1,11-cyclization in the presence of an alternative metal ion
the three cytochrome P450 monooxygenases CYP5344B1, CYP5348E1, and CYP5348J3 show substantial activities against DELTA6-protoilludene catalyzing the hydroxylation reaction of DELTA6-protoilludene to produce DELTA6-protoilludene-8-ol and DELTA6-protoilludene-5-ol
site-directed mutagenesis of Omp7, compared to wild-type, the mutant shows moderate reduction of activity with Mg2+ and Mn2+, slight to moderate reduction with Co2+, slightly reduced activity with Ni2+, and strong reduction of activity with Ca2+
site-directed mutagenesis of Omp7, compared to wild-type, the mutant shows moderate reduction of activity with Co2+, Mg2+, and Mn2+, slightly reduced activity with Ni2+, and strong reduction of activity with Ca2+
site-directed mutagenesis of Omp6, compared to wild-type, the mutant shows moderate reduction of activity with Mg2+ and Ni2+, slight to moderate reduction with Co2+, slightly reduced activity with Mn2+, and very strong reduction of activity with Ca2+
site-directed mutagenesis of Stehi73029, compared to wild-type, the mutant shows moderately reduced activity with Mg2+, slight to moderate reduction of activity with Mn2+, strong reduction of activity with Co2+, and no activity with Ca2+ and Ni2+
site-directed mutagenesis of Stehi73029, compared to wild-type, the mutant shows slight to moderate reduction of activity with Mg2+, strongly reduced activity with Ca2+ and Mn2+, very strong reduction of activity with Co2+, and no activity with Ni2+
site-directed mutagenesis of Stehi73029, compared to wild-type, the mutant shows moderate reduction of activity with Mg2+, strong reduction with Ca2+, Co2+, and Mn2+, and very strong reduction with Ni2+
site-directed mutagenesis of Stehi73029, compared to wild-type, the mutant shows slight to moderate reduction of activity with Mg2+, and no activity with Ca2+, Mn2+, Ni2+, and Co2+
site-directed mutagenesis of Stehi73029, compared to wild-type, the mutant shows slight reduction of activity with Mg2+, moderately reduced activity with Ca2+ and Mn2+, and no activity with Ni2+ and Co2+
protoilludene is successfully produced in Escherichia coli by overexpression of a hybrid exogenous mevalonate pathway, endogenous farnesyl diphosphate synthase (IspA), and protoilludene synthase (OMP7) of Omphalotus olearius
coexpression of protoilludene synthase PpSTS-08 and diverse PpCYPs in Saccharomyces cerevisiae by chromosomal integration of the PpSTS-08 expression cassette and subsequent transformation with expression plasmids of 184 PpCYPs previously constructed resulting in 184 distinct transformants that give simultaneous expression of PpSTS-08 with each of the PpCYPs
recombinant wild-type and mutant His6-tagged Omp6 enzymes from Escherichia coli by affinity chromatography and dialysis against EDTA to eliminate metal ions
recombinant wild-type and mutant His6-tagged Omp7 enzymes from Escherichia coli by affinity chromatography and dialysis against EDTA to eliminate metal ions
recombinant wild-type and mutant His6-tagged Stehi73029 enzymes from Escherichia coli by affinity chromatography and dialysis against EDTA to eliminate metal ions
PpSTS-08, coexpression of protoilludene synthase PpSTS-08 and diverse PpCYPs in Saccharomyces cerevisiae by chromosomal integration of the PpSTS-08 expression cassette and subsequent transformation with expression plasmids of 184 PpCYPs previously constructed resulting in 184 distinct transformants that give simultaneous expression of PpSTS-08 with each of the PpCYPs
protoilludene is a valuable sesquiterpene and serves as a precursor for several medicinal compounds and antimicrobial chemicals. It can be synthesized by heterologous overexpression of protoilludene synthase in Escherichia coli with coexpression of mevalonate or methylerythritol-phosphate pathway enzymes, and farnesyl diphosphate synthase as a cell factory for protoilludene production
Engels, B.; Heinig, U.; Grothe, T.; Stadler, M.; Jennewein, S.
Cloning and characterization of an Armillaria gallica cDNA encoding protoilludene synthase, which catalyzes the first committed step in the synthesis of antimicrobial melleolides
Quin, M.; Flynn, C.; Wawrzyn, G.; Choudhary, S.; Schmidt-Dannert, C.
Mushroom hunting by using bioinformatics: Application of a predictive framework facilitates the selective identification of sesquiterpene synthases in Basidiomycota
Insight into metabolic diversity of the brown-rot basidiomycete Postia placenta responsible for sesquiterpene biosynthesis semi-comprehensive screening of cytochrome P450 monooxygenase involved in protoilludene metabolism