BRENDA - Enzyme Database show
show all sequences of 1.14.13.183

Optimization of a cytochrome P450 oxidation system for enhancing protopanaxadiol production in Saccharomyces cerevisiae

Zhao, F.; Bai, P.; Liu, T.; Li, D.; Zhang, X.; Lu, W.; Yuan, Y.; Biotechnol. Bioeng. 113, 1787-1795 (2016)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
recombinant expression of wild-type and mutant fusion proteins in Saccharomyces cerevisiae strain W303-1a. coexpression of wild-type enzyme with dammarenediol-II synthase from Panax ginseng and a Arabidopsis thaliana NADPH-cytochrome P450 reductase (ATR1). The low metabolic flux through PPDS results in a low productivity of protopanaxadiol, but is increased with the fusion enzymes
Panax ginseng
Engineering
Amino acid exchange
Commentary
Organism
additional information
to increase the low metabolic flux through PPDS and increase the productivity of protopanaxadiol production, enzyme PPDS is modified through transmembrane domain truncation and construction of self-sufficient PPDS-ATR1 fusion proteins. The fusion enzymes exhibit approximately 4.5fold increase in catalytic activity and 71.1% increase in protopanaxadiol production compared with PPDS and ATR1 co-expression. The engineered yeast carrying fusion protein effectively converts 96.8% of dammarenediol-II into protopanaxadiol, bioreactor in fed-batch fermentation. Construction of four fusion proteins named as PPDS-linker1-46tATR1, PPDSlinker2-46tATR1, PPDS-linker3-46tATR1, and PPDS-nolinker-46tATR1, respectively. In addition, 31tPPDS heme domain and 46tATR1 reductase domain are linked by polypeptide GSTSSGSG. Method optimization, overview. Evaluation of the oxidative stress in yeast cells induced by co-expression of PPDS and ATR1
Panax ginseng
Localization
Localization
Commentary
Organism
GeneOntology No.
Textmining
endoplasmic reticulum membrane
the enzyme contains a transmembrane domain
Panax ginseng
5789
-
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
dammarenediol-II + NADPH + H+ + O2
Panax ginseng
-
protopanaxadiol + NADP+ + H2O
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Panax ginseng
H2DH16
-
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
dammarenediol-II + NADPH + H+ + O2
-
744549
Panax ginseng
protopanaxadiol + NADP+ + H2O
-
-
-
?
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at
Panax ginseng
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.4
-
assay at
Panax ginseng
Cofactor
Cofactor
Commentary
Organism
Structure
cytochrome P450
-
Panax ginseng
NADPH
-
Panax ginseng
Cloned(Commentary) (protein specific)
Commentary
Organism
recombinant expression of wild-type and mutant fusion proteins in Saccharomyces cerevisiae strain W303-1a. coexpression of wild-type enzyme with dammarenediol-II synthase from Panax ginseng and a Arabidopsis thaliana NADPH-cytochrome P450 reductase (ATR1). The low metabolic flux through PPDS results in a low productivity of protopanaxadiol, but is increased with the fusion enzymes
Panax ginseng
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
cytochrome P450
-
Panax ginseng
NADPH
-
Panax ginseng
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
additional information
to increase the low metabolic flux through PPDS and increase the productivity of protopanaxadiol production, enzyme PPDS is modified through transmembrane domain truncation and construction of self-sufficient PPDS-ATR1 fusion proteins. The fusion enzymes exhibit approximately 4.5fold increase in catalytic activity and 71.1% increase in protopanaxadiol production compared with PPDS and ATR1 co-expression. The engineered yeast carrying fusion protein effectively converts 96.8% of dammarenediol-II into protopanaxadiol, bioreactor in fed-batch fermentation. Construction of four fusion proteins named as PPDS-linker1-46tATR1, PPDSlinker2-46tATR1, PPDS-linker3-46tATR1, and PPDS-nolinker-46tATR1, respectively. In addition, 31tPPDS heme domain and 46tATR1 reductase domain are linked by polypeptide GSTSSGSG. Method optimization, overview. Evaluation of the oxidative stress in yeast cells induced by co-expression of PPDS and ATR1
Panax ginseng
Localization (protein specific)
Localization
Commentary
Organism
GeneOntology No.
Textmining
endoplasmic reticulum membrane
the enzyme contains a transmembrane domain
Panax ginseng
5789
-
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
dammarenediol-II + NADPH + H+ + O2
Panax ginseng
-
protopanaxadiol + NADP+ + H2O
-
-
?
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
dammarenediol-II + NADPH + H+ + O2
-
744549
Panax ginseng
protopanaxadiol + NADP+ + H2O
-
-
-
?
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at
Panax ginseng
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.4
-
assay at
Panax ginseng
Other publictions for EC 1.14.13.183
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)
744549
Zhao
Optimization of a cytochrome ...
Panax ginseng
Biotechnol. Bioeng.
113
1787-1795
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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1
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746053
Chun
Production of the dammarene s ...
Panax ginseng
Plant Cell Rep.
34
1551-1560
2015
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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
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1
2
2
1
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745011
Wang
The isolation and characteriz ...
Panax quinquefolius
Funct. Integr. Genomics
14
545-557
2014
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1
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-
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1
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1
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1
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