Enzymic formation of sesquiterpenes

Cane, David E.

doi:10.1021/cr00105a002

Cited by 461 publications

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“…By using the structure of the trichodiene synthase-pyrophosphate complex and the mechanism of trichodiene formation (3,(34)(35)(36) as a guide, we have constructed a structure-based model of the trichodiene synthase reaction (Fig. 5).…”

Section: Resultsmentioning

confidence: 99%

“…catalyze the conversion of a universal substrate, farnesyl diphosphate, into more than 300 known terpene cyclization products with different structures and stereochemistries (1)(2)(3)(4)(5). It is striking that such broad product diversity is achieved by a structurally homologous group of enzymes, each of which typically has evolved to catalyze a single cyclization reaction with exquisite structural and stereochemical precision.…”

Section: S Esquiterpene Synthases (Also Known As Terpenoid Cyclases)mentioning

confidence: 99%

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Structure of trichodiene synthase from Fusarium sporotrichioides provides mechanistic inferences on the terpene cyclization cascade

Rynkiewicz

Cane

Christianson

2001

Proc. Natl. Acad. Sci. U.S.A.

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The x-ray crystal structure of recombinant trichodiene synthase from Fusarium sporotrichioides has been determined to 2.5-Å resolution, both unliganded and complexed with inorganic pyrophosphate. This reaction product coordinates to three Mg 2؉ ions near the mouth of the active site cleft. A comparison of the liganded and unliganded structures reveals a ligand-induced conformational change that closes the mouth of the active site cleft. Binding of the substrate farnesyl diphosphate similarly may trigger this conformational change, which would facilitate catalysis by protecting reactive carbocationic intermediates in the cyclization cascade. Trichodiene synthase also shares significant structural similarity with other sesquiterpene synthases despite a lack of significant sequence identity. This similarity indicates divergence from a common ancestor early in the evolution of terpene biosynthesis.

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Section: Resultsmentioning

confidence: 99%

Section: S Esquiterpene Synthases (Also Known As Terpenoid Cyclases)mentioning

confidence: 99%

Structure of trichodiene synthase from Fusarium sporotrichioides provides mechanistic inferences on the terpene cyclization cascade

Rynkiewicz

Cane

Christianson

2001

Proc. Natl. Acad. Sci. U.S.A.

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254

306

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“…Taken together, these findings show that the active site of trichodiene synthase is sufficiently flexible to accommodate bulkier and electronically-diverse substrates and intermediates, which could indicate additional potential for the biosynthetic utility of this terpenoid cyclase. Keywords terpenoid synthase; farnesyl diphosphate; sesquiterpene; protein crystallography Sesquiterpene synthases catalyze the cyclization of the universal acyclic precursor, farnesyl diphosphate (FPP) 1 , to form one or more of over 300 known monocyclic, bicyclic, and tricyclic sesquiterpenes with a wide variety of structures and stereochemistry [1][2][3][4][5]. All these reactions are known to involve a cascade of carbocationic intermediates, and the controlled manipulation of these highly reactive intermediates gives rise to impressive product diversity.…”

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confidence: 99%

Exploring biosynthetic diversity with trichodiene synthase

Vedula

Zhao

Coates

et al. 2007

Archives of Biochemistry and Biophysics

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Trichodiene synthase is a terpenoid cyclase that catalyzes the cyclization of farnesyl diphosphate (FPP) to form the bicyclic sesquiterpene hydrocarbon trichodiene (89%), at least five sesquiterpene side products (11%), and inorganic pyrophosphate (PP i ). Incubation of trichodiene synthase with 2-fluorofarnesyl diphosphate or 4-methylfarnesyl diphosphate similarly yields sesquiterpene mixtures despite the electronic effects or steric bulk introduced by substrate derivatization. The versatility of the enzyme is also demonstrated in the 2.85 Å resolution X-ray crystal structure of the complex with Mg 2+ 3 -PP i and the benzyl triethylammonium cation, which is a bulkier mimic of the bisabolyl carbocation intermediate in catalysis. Taken together, these findings show that the active site of trichodiene synthase is sufficiently flexible to accommodate bulkier and electronically-diverse substrates and intermediates, which could indicate additional potential for the biosynthetic utility of this terpenoid cyclase. Keywords terpenoid synthase; farnesyl diphosphate; sesquiterpene; protein crystallography Sesquiterpene synthases catalyze the cyclization of the universal acyclic precursor, farnesyl diphosphate (FPP) 1 , to form one or more of over 300 known monocyclic, bicyclic, and tricyclic sesquiterpenes with a wide variety of structures and stereochemistry [1][2][3][4][5]. All these reactions are known to involve a cascade of carbocationic intermediates, and the controlled manipulation of these highly reactive intermediates gives rise to impressive product diversity. Typically, the active site of a terpene cyclase serves as a template that chaperones the conformations and ⋆ This work was supported by NIH grants GM56838 (D.W.C.), GM30301 (D.E.C.), and GM13956 (R.M.C.).⋆⋆ Atomic coordinates of wild-type trichodiene synthase complexed with BTAC and with inorganic pyrophosphate, resulting from the reaction with 2-fluorofarnesyl diphosphate, have been deposited in the Research Collaboratory for Structural Bioinformatics (http:// www.rcsb.org/pdb) with the following accession codes: 2Q9Y and 2Q9Z, respectively. c Current address: Takeda San Diego, Inc., 10410 Science Center Drive, San Diego, California 92121, USA * Corresponding author. Fax: +1 215-573-2201. E-mail address: chris@sas.upenn.edu.Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. 1 Abbreviations: FPP, farnesyl diphosphate; PP i , inorganic pyrophosphate; 2FFPP, 2-fluorofarnesyl diphosphate; 4M-FPP, 4-methylfarnesyl diphosphate; BTAC, benzyl triethylammonium cation; GC, gas chromatography; MS, mass spectroscopy; r.m....

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“…Keywords enzyme kinetics; protein crystallography; terpenoid cyclase; farnesyl diphosphate Sesquiterpene synthases catalyze the metal ion-dependent cyclization of the universal acyclic substrate, farnesyl diphosphate (FPP), to form one of more than 300 known monocyclic, bicyclic, or tricyclic hydrocarbon or alcohol products of widely varied structure and stereochemistry [1][2][3][4][5]. These cyclic products represent branch points in terpenoid biosynthesis.…”

Section: Introductionmentioning

confidence: 99%

“…Each catalytically active mutant generates a different distribution of sesquiterpene products, and three newly detected sesquiterpenes are identified. In addition, the kinetic and structural properties of the Y295F mutant of trichodiene synthase were found to be similar to those of the wild-type enzyme, thereby ruling out a proposed role for Y295 in catalysis.Keywords enzyme kinetics; protein crystallography; terpenoid cyclase; farnesyl diphosphate Sesquiterpene synthases catalyze the metal ion-dependent cyclization of the universal acyclic substrate, farnesyl diphosphate (FPP), to form one of more than 300 known monocyclic, bicyclic, or tricyclic hydrocarbon or alcohol products of widely varied structure and stereochemistry [1][2][3][4][5]. These cyclic products represent branch points in terpenoid biosynthesis.…”

mentioning

confidence: 99%

Structural and mechanistic analysis of trichodiene synthase using site-directed mutagenesis: Probing the catalytic function of tyrosine-295 and the asparagine-225/serine-229/glutamate-233–Mg2+B
Vedula
¹
,
Jiang
²
,
Zakharian
³

et al. 2008
Archives of Biochemistry and Biophysics
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Trichodiene synthase from Fusarium sporotrichioides contains two metal ion-binding motifs required for the cyclization of farnesyl diphosphate: the "aspartate-rich" motif D 100 DXX(D/E) that coordinates to Mg 2+ A and Mg 2+ C , and the "NSE/DTE" motif N 225 DXXSXXXE that chelates Mg 2+ B (boldface indicates metal ion ligands). Here, we report steady-state kinetic parameters, product array analyses, and X-ray crystal structures of trichodiene synthase mutants in which the fungal NSE motif is progressively converted into a plant-like DDXXTXXXE motif, resulting in a degradation in both steady-state kinetic parameters and product specificity. Each catalytically active mutant generates a different distribution of sesquiterpene products, and three newly detected sesquiterpenes are identified. In addition, the kinetic and structural properties of the Y295F mutant of trichodiene synthase were found to be similar to those of the wild-type enzyme, thereby ruling out a proposed role for Y295 in catalysis.Keywords enzyme kinetics; protein crystallography; terpenoid cyclase; farnesyl diphosphate Sesquiterpene synthases catalyze the metal ion-dependent cyclization of the universal acyclic substrate, farnesyl diphosphate (FPP), to form one of more than 300 known monocyclic, bicyclic, or tricyclic hydrocarbon or alcohol products of widely varied structure and stereochemistry [1][2][3][4][5]. These cyclic products represent branch points in terpenoid biosynthesis. Further modifications of cyclic sesquiterpenes and diterpenes involving downstream enzymes such as the cytochrome P450s of Artemisia annua and Taxus brevifolia, or the Fe(II)-α-I This work was supported by NIH grants GM56838 (D.W.C.) and GM30301 (D.E.C.).II Atomic coordinates and structure factors for N225D trichodiene synthase, the N225D trichodiene synthase-Mg 2+ 3 -PPi complex, N225D/S229T trichodiene synthase, Y295F trichodiene synthase, and the Y295F trichodiene synthase-Mg 2+ 3 -PPi complex have been deposited in the Research Collaboratory for Structural Bioinformatics (http://www.rcsb.org/pdb) with the following accession codes: 2PS4, 2PS5, 2PS6, 2PS7, and 2PS8, respectively. *Corresponding author. Fax: +1 215-573-2201. E-mail: chris@sas.upenn.edu. Fax: +1 401-863-9368. E-mail: David_Cane@Brown.edu. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. To date, the dissection of the NSE/DTE-Mg 2+ B coordination motif in terpene cyclases by sitedirected mutagenesis has not been accompanied by X-ray crystal structure analysis. Accordingly, we now report the first exploration of structure-function relationships for this ...

show abstract

Enzymic formation of sesquiterpenes

Cited by 461 publications

References 35 publications

Structure of trichodiene synthase from Fusarium sporotrichioides provides mechanistic inferences on the terpene cyclization cascade

Structure of trichodiene synthase from Fusarium sporotrichioides provides mechanistic inferences on the terpene cyclization cascade

Exploring biosynthetic diversity with trichodiene synthase

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