BRENDA - Enzyme Database show
show all sequences of 4.2.3.104

Heterologous expression, purification, and biochemical characterization of alpha-humulene synthase from Zingiber zerumbet Smith

Alemdar, S.; Hartwig, S.; Frister, T.; Koenig, J.C.; Scheper, T.; Beutel, S.; Appl. Biochem. Biotechnol. 178, 474-489 (2016)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
gene ZSS1 or hum, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) under control of the lac promoter
Zingiber zerumbet
Engineering
Amino acid exchange
Commentary
Organism
additional information
alpha-humulene production using the engineered Escherichia coli BL21(DE3) strain in a 2-l bioreactor cultivation, method optimization, overview. Coordinating Cu2+ ions to the NTA ligands in affinity chromatography results in a high non-specific binding of endogenous host cell proteins due to the high affinity of Cu2+ to histidine groups. Zn2+-IMAC shows much less impurities in eluted protein fractions which is consistent with the fact that Zn2+ ions are less affine to histidine groups. The recovery rate of 80 % of the protein of interest using Zn2+ ions is the lowest compared to all other metal ions tested. Ni2+- and Co2+-IMAC seem to be similar in product purity (60% and 63%) and recovery (93% and 95%). As Co2+-IMAC shows the best purity to recovery ratio, further purification is done using Co2+-IMAC
Zingiber zerumbet
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
Michaelis-Menten kinetics, overview
Zingiber zerumbet
0.033
-
(2E,6E)-farnesyl diphosphate
pH 7.0, 38C, recombinant enzyme
Zingiber zerumbet
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
(2E,6E)-farnesyl diphosphate
Zingiber zerumbet
-
alpha-humulene + diphosphate
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Zingiber zerumbet
B1B1U3
-
-
Purification (Commentary)
Commentary
Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3)by cobalt affinity chromatography, dialysis, and ultrafiltration, to homogeneity
Zingiber zerumbet
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(2E,6E)-farnesyl diphosphate
-
746789
Zingiber zerumbet
alpha-humulene + diphosphate
-
-
-
?
additional information
the recombinant alpha-humulene synthase produces 93.45% alpha-humulene and 5.5% beta-caryophyllene. Product determination by gas chromatography-flame ionization detection (GC-FID)
746789
Zingiber zerumbet
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
?
x * 64500, about, sequence calculation
Zingiber zerumbet
Temperature Optimum [C]
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
38
-
recombinant His-tagged enzyme
Zingiber zerumbet
Temperature Range [C]
Temperature Minimum [C]
Temperature Maximum [C]
Commentary
Organism
32
44
activity range, profile overview, recombinant His-tagged enzyme
Zingiber zerumbet
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.8
-
(2E,6E)-farnesyl diphosphate
pH 7.0, 38C, recombinant enzyme
Zingiber zerumbet
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
recombinant His-tagged enzyme
Zingiber zerumbet
pH Range
pH Minimum
pH Maximum
Commentary
Organism
6
9
activity range, profile overview, recombinant His-tagged enzyme
Zingiber zerumbet
pI Value
Organism
Commentary
pI Value Maximum
pI Value
Zingiber zerumbet
recombinant His-tagged enzyme, isoelectric focusing
-
5.61
Zingiber zerumbet
sequence calculation
-
5.65
Cloned(Commentary) (protein specific)
Commentary
Organism
gene ZSS1 or hum, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) under control of the lac promoter
Zingiber zerumbet
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
additional information
alpha-humulene production using the engineered Escherichia coli BL21(DE3) strain in a 2-l bioreactor cultivation, method optimization, overview. Coordinating Cu2+ ions to the NTA ligands in affinity chromatography results in a high non-specific binding of endogenous host cell proteins due to the high affinity of Cu2+ to histidine groups. Zn2+-IMAC shows much less impurities in eluted protein fractions which is consistent with the fact that Zn2+ ions are less affine to histidine groups. The recovery rate of 80 % of the protein of interest using Zn2+ ions is the lowest compared to all other metal ions tested. Ni2+- and Co2+-IMAC seem to be similar in product purity (60% and 63%) and recovery (93% and 95%). As Co2+-IMAC shows the best purity to recovery ratio, further purification is done using Co2+-IMAC
Zingiber zerumbet
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
Michaelis-Menten kinetics, overview
Zingiber zerumbet
0.033
-
(2E,6E)-farnesyl diphosphate
pH 7.0, 38C, recombinant enzyme
Zingiber zerumbet
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
(2E,6E)-farnesyl diphosphate
Zingiber zerumbet
-
alpha-humulene + diphosphate
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3)by cobalt affinity chromatography, dialysis, and ultrafiltration, to homogeneity
Zingiber zerumbet
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(2E,6E)-farnesyl diphosphate
-
746789
Zingiber zerumbet
alpha-humulene + diphosphate
-
-
-
?
additional information
the recombinant alpha-humulene synthase produces 93.45% alpha-humulene and 5.5% beta-caryophyllene. Product determination by gas chromatography-flame ionization detection (GC-FID)
746789
Zingiber zerumbet
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
?
x * 64500, about, sequence calculation
Zingiber zerumbet
Temperature Optimum [C] (protein specific)
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
38
-
recombinant His-tagged enzyme
Zingiber zerumbet
Temperature Range [C] (protein specific)
Temperature Minimum [C]
Temperature Maximum [C]
Commentary
Organism
32
44
activity range, profile overview, recombinant His-tagged enzyme
Zingiber zerumbet
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.8
-
(2E,6E)-farnesyl diphosphate
pH 7.0, 38C, recombinant enzyme
Zingiber zerumbet
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
recombinant His-tagged enzyme
Zingiber zerumbet
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
6
9
activity range, profile overview, recombinant His-tagged enzyme
Zingiber zerumbet
pI Value (protein specific)
Organism
Commentary
pI Value Maximum
pI Value
Zingiber zerumbet
recombinant His-tagged enzyme, isoelectric focusing
-
5.61
Zingiber zerumbet
sequence calculation
-
5.65
General Information
General Information
Commentary
Organism
metabolism
the enzyme is involved in the zerumbone biosynthesis in Zingiber zerumbet, proposed pathway and mechanism, overview
Zingiber zerumbet
physiological function
alpha-humulene is a common sesquiterpene widely distributed in plants, the synthase from Zingiber zerumbet is the only known sesquiterpene synthase to date catalyzing the formation of alpha-humulene as major product
Zingiber zerumbet
General Information (protein specific)
General Information
Commentary
Organism
metabolism
the enzyme is involved in the zerumbone biosynthesis in Zingiber zerumbet, proposed pathway and mechanism, overview
Zingiber zerumbet
physiological function
alpha-humulene is a common sesquiterpene widely distributed in plants, the synthase from Zingiber zerumbet is the only known sesquiterpene synthase to date catalyzing the formation of alpha-humulene as major product
Zingiber zerumbet
Other publictions for EC 4.2.3.104
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)
746751
Krieg
CO2 to terpenes autotrophic a ...
Cupriavidus necator
Angew. Chem. Int. Ed. Engl.
57
1879-1882
2018
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1
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748561
Zhang
Production of sesquiterpenoid ...
Zingiber zerumbet
Metab. Eng.
49
28-35
2018
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1
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1
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1
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747656
Alemdar
-
Bioproduction of alpha-humule ...
Zingiber zerumbet
Eng. Life Sci.
17
900-907
2017
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1
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4
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1
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1
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1
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1
1
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746789
Alemdar
Heterologous expression, puri ...
Zingiber zerumbet
Appl. Biochem. Biotechnol.
178
474-489
2016
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1
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1
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1
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1
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2
1
1
1
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1
1
1
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2
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2
2
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748440
Kumeta
Characterization of alpha-hum ...
Aquilaria crassna
J. Nat. Med.
70
452-459
2016
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1
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1
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3
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3
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1
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6
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3
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6
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1
2
6
3
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748552
Sonntag
Engineering Methylobacterium ...
Zingiber zerumbet
Metab. Eng.
32
82-94
2015
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1
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1
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1
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730575
Yang
Isolation and characterization ...
Gossypium hirsutum
Phytochemistry
96
46-56
2013
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-
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1
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2
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5
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7
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2
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1
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716562
Bleeker
RNA-seq discovery, functional ...
Solanum habrochaites
Plant Mol. Biol.
77
323-336
2011
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717468
Yu
Zingiber zerumbet CYP71BA1 cat ...
Zingiber zerumbet
Cell. Mol. Life Sci.
68
1033-1040
2011
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1
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1
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1
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1
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716459
Koepke
Species-specific responses of ...
Pinus sylvestris
Phytochemistry
71
909-917
2010
-
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8
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718233
Schilmiller
Studies of a biochemical facto ...
Solanum lycopersicum
Plant Physiol.
153
1212-1223
2010
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1
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690593
Harada
Efficient synthesis of functio ...
Zingiber zerumbet
Appl. Microbiol. Biotechnol.
81
915-925
2008
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1
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6
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1
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718244
Yu
Molecular cloning and function ...
Zingiber zerumbet
Planta
227
1291-1299
2008
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1
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1
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1
1
1
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1
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713266
van Der Hoeven
Genetic control and evolution ...
no activity in Solanum habrochaites, Solanum lycopersicum
Plant Cell
12
2283-2294
2000
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4
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