BRENDA - Enzyme Database show
show all sequences of 4.2.3.119

Mutational analysis of a monoterpene synthase reaction: altered catalysis through directed mutagenesis of (-)-pinene synthase from Abies grandis

Hyat, D.C.; Croteau, R.; Arch. Biochem. Biophys. 439, 222-233 (2005)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expression in Escherichia coli
Abies grandis
Crystallization (Commentary)
Crystallization
Organism
modeling of amino acid sequence onto the crystal structures of tobacco 5-epi-aristolochene synthase and bornyl diphosphate synthase and comparison with (-)-camphene synthase
Abies grandis
Engineering
Amino acid exchange
Commentary
Organism
C372S
replacement with corresponding residue of (-)-camphene synthase, 97% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene, while the levels of total pinenes remains relatively constant
Abies grandis
C372S/C480S
replacement with corresponding residue of (-)-camphene synthase, 72% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene
Abies grandis
C372S/F597W
replacement with corresponding residue of (-)-camphene synthase, 100% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene
Abies grandis
C372S/F597W/S485C/F597W
replacement with corresponding residue of (-)-camphene synthase, 99% of wild-type activity. Mutant produces about 80%(-)-alpha-pinene and 10% (-)-beta-pinene
Abies grandis
C372S/S485C
replacement with corresponding residue of (-)-camphene synthase, 92% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene
Abies grandis
C480S
replacement with corresponding residue of (-)-camphene synthase, 97% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene, while the levels of total pinenes remains relatively constant
Abies grandis
C480S/F597W
replacement with corresponding residue of (-)-camphene synthase, 7% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene
Abies grandis
C480S/S485C
replacement with corresponding residue of (-)-camphene synthase, 70% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene
Abies grandis
F597W
replacement with corresponding residue of (-)-camphene synthase, 73% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene, while the levels of total pinenes remains relatively constant
Abies grandis
additional information
replacement of selected amino acid residues in (-)-pinene synthase with the corresponding residues from (-)-camphene synthase in an effort to identify the amino acids responsible for the catalytic diVerences. The approach produces an enzyme in which more than half of the product is channeled through an alternative pathway. Several (-)-pinene synthase to (-)-camphene synthase amino acid substitutions are necessary before catalysis is significantly altered
Abies grandis
S485C
replacement with corresponding residue of (-)-camphene synthase, 100% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene, while the levels of total pinenes remains relatively constant
Abies grandis
S485C/F597W
replacement with corresponding residue of (-)-camphene synthase, 68% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene
Abies grandis
Localization
Localization
Commentary
Organism
GeneOntology No.
Textmining
chloroplast
-
Abies grandis
9507
-
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Abies grandis
O24475
-
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
geranyl diphosphate
-
663770
Abies grandis
(-)-alpha-pinene + diphosphate
products are 29% (-)-alpha-pinene, 63% (-)-beta-pinene, 1.8% myrcene, 3.6% limonene
-
-
?
Cloned(Commentary) (protein specific)
Commentary
Organism
expression in Escherichia coli
Abies grandis
Crystallization (Commentary) (protein specific)
Crystallization
Organism
modeling of amino acid sequence onto the crystal structures of tobacco 5-epi-aristolochene synthase and bornyl diphosphate synthase and comparison with (-)-camphene synthase
Abies grandis
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
C372S
replacement with corresponding residue of (-)-camphene synthase, 97% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene, while the levels of total pinenes remains relatively constant
Abies grandis
C372S/C480S
replacement with corresponding residue of (-)-camphene synthase, 72% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene
Abies grandis
C372S/F597W
replacement with corresponding residue of (-)-camphene synthase, 100% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene
Abies grandis
C372S/F597W/S485C/F597W
replacement with corresponding residue of (-)-camphene synthase, 99% of wild-type activity. Mutant produces about 80%(-)-alpha-pinene and 10% (-)-beta-pinene
Abies grandis
C372S/S485C
replacement with corresponding residue of (-)-camphene synthase, 92% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene
Abies grandis
C480S
replacement with corresponding residue of (-)-camphene synthase, 97% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene, while the levels of total pinenes remains relatively constant
Abies grandis
C480S/F597W
replacement with corresponding residue of (-)-camphene synthase, 7% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene
Abies grandis
C480S/S485C
replacement with corresponding residue of (-)-camphene synthase, 70% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene
Abies grandis
F597W
replacement with corresponding residue of (-)-camphene synthase, 73% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene, while the levels of total pinenes remains relatively constant
Abies grandis
additional information
replacement of selected amino acid residues in (-)-pinene synthase with the corresponding residues from (-)-camphene synthase in an effort to identify the amino acids responsible for the catalytic diVerences. The approach produces an enzyme in which more than half of the product is channeled through an alternative pathway. Several (-)-pinene synthase to (-)-camphene synthase amino acid substitutions are necessary before catalysis is significantly altered
Abies grandis
S485C
replacement with corresponding residue of (-)-camphene synthase, 100% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene, while the levels of total pinenes remains relatively constant
Abies grandis
S485C/F597W
replacement with corresponding residue of (-)-camphene synthase, 68% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene
Abies grandis
Localization (protein specific)
Localization
Commentary
Organism
GeneOntology No.
Textmining
chloroplast
-
Abies grandis
9507
-
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
geranyl diphosphate
-
663770
Abies grandis
(-)-alpha-pinene + diphosphate
products are 29% (-)-alpha-pinene, 63% (-)-beta-pinene, 1.8% myrcene, 3.6% limonene
-
-
?
Other publictions for EC 4.2.3.119
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
747323
Tashiro
Tweezing the cofactor prefere ...
Pinus taeda
Biosci. Biotechnol. Biochem.
82
1058-1061
2018
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4
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2
2
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747887
Wang
An integrative volatile terpe ...
Wurfbainia villosa
Front. Plant Sci.
9
846
2018
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1
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3
3
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746601
Tashiro
Bacterial production of pinen ...
Pinus taeda
ACS Synth. Biol.
5
1011-1020
2016
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1
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3
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1
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1
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3
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2
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747387
Ma
Characterization of a monoter ...
Paeonia lactiflora
Biotechnol. Lett.
38
1213-1219
2016
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1
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1
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2
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729473
Kang
Biosynthesis of pinene from gl ...
Pinus taeda
Biotechnol. Lett.
36
2069-2077
2014
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1
1
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729506
Hall
Transcriptome resources and fu ...
Pinus banksiana, Pinus contorta
BMC Plant Biol.
13
80
2013
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2
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730575
Yang
Isolation and characterization ...
Gossypium hirsutum
Phytochemistry
96
46-56
2013
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730632
Huang
Identification and characteriz ...
Gossypium hirsutum
Plant Physiol. Biochem.
73
302-308
2013
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663770
Hyat
Mutational analysis of a monot ...
Abies grandis
Arch. Biochem. Biophys.
439
222-233
2005
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1
1
12
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694598
Huber
Characterization of four terpe ...
Pseudotsuga menziesii
Phytochemistry
66
1427-1439
2005
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649494
Katoh S.;Hyatt D.;Croteau R.
Altering product outcome in Ab ...
Abies grandis
Arch. Biochem. Biophys.
425
65-76
2004
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717151
Phillips
cDNA isolation, functional exp ...
Pinus taeda
Arch. Biochem. Biophys.
411
267-276
2003
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718229
McKay
Insect attack and wounding ind ...
Picea sitchensis
Plant Physiol.
133
368-378
2003
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653533
Lu
Cloning and functional charact ...
Artemisia annua
Plant Physiol.
130
477-486
2002
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701897
Bohlmann
cDNA cloning, characterization ...
Abies grandis
Arch. Biochem. Biophys.
368
232-243
1999
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717150
Phillips
Monoterpene synthases of loblo ...
Pinus taeda
Arch. Biochem. Biophys.
372
197-204
1999
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210753
Bohlmann
Monoterpene synthases from gra ...
Abies grandis
J. Biol. Chem.
272
21784-21792
1997
1
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717148
McGeady
Isolation and characterization ...
Salvia officinalis
Arch. Biochem. Biophys.
317
149-155
1995
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4
2
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6
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210755
Wagschal
Monoterpene biosynthesis: Isot ...
Salvia officinalis
Arch. Biochem. Biophys.
308
477-487
1994
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210762
Pyun
Stereochemistry of the proton ...
Salvia officinalis
Arch. Biochem. Biophys.
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488-496
1994
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210760
Lewinsohn
Wound-inducible pinene cyclase ...
Abies grandis
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167-173
1992
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210754
Gijzen
Characterization of the consti ...
Abies grandis
Arch. Biochem. Biophys.
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267-273
1991
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210758
Croteau
Biosynthesis of monoterpenes: ...
Salvia officinalis
J. Biol. Chem.
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15309-15315
1989
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4
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210757
Croteau
Biosynthesis of monoterpenes: ...
Salvia officinalis
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10063-10071
1988
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4
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9
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9
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210756
Croteau
Isotopically sensitive branchi ...
Salvia officinalis
Biochemistry
26
5383-5389
1987
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717749
Gambliel
Pinene cyclases I and II. Two ...
Salvia officinalis
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740-748
1984
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3
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