BRENDA - Enzyme Database
show all sequences of 1.1.1.365

Functional characterization of D-galacturonic acid reductase, a key enzyme of the ascorbate biosynthesis pathway, from Euglena gracilis

Ishikawa, T.; Masumoto, I.; Iwasa, N.; Nishikawa, H.; Sawa, Y.; Shibata, H.; Nakamura, A.; Yabuta, Y.; Shigeoka, S.; Biosci. Biotechnol. Biochem. 70, 2720-2726 (2006)

Data extracted from this reference:

Activating Compound
Activating Compound
Commentary
Organism
Structure
H2O2
the enzyme is activated by 0.1 mM H2O2
Euglena gracilis
additional information
not influenced by EDTA, BCS, dithiothreiol, , and p-chloromercuribenzoate, L-GalL, and AsA
Euglena gracilis
Inhibitors
Inhibitors
Commentary
Organism
Structure
N-ethylmaleimide
84% residual activity at 1 mM
Euglena gracilis
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0625
-
NADPH
in 50 mM Tris-HCl (pH 7.2), at 25°C
Euglena gracilis
3.79
-
D-galacturonate
in 50 mM Tris-HCl (pH 7.2), at 25°C
Euglena gracilis
4.67
-
D-glucuronate
in 50 mM Tris-HCl (pH 7.2), at 25°C
Euglena gracilis
8.48
-
D-xylose
in 50 mM Tris-HCl (pH 7.2), at 25°C
Euglena gracilis
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Cu2+
135.3% activity at 5 mM
Euglena gracilis
additional information
not influenced by Ca2+, Co2+, Mn2+, Ni2+, Zn2+, and Fe2+
Euglena gracilis
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
38000
-
1 * 38000, SDS-PAGE
Euglena gracilis
39000
-
gel filtration
Euglena gracilis
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
D-galacturonate + NADPH + H+
Euglena gracilis
-
L-galactonate + NADP+
-
-
ir
D-galacturonate + NADPH + H+
Euglena gracilis Z
-
L-galactonate + NADP+
-
-
ir
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Euglena gracilis
-
-
-
Euglena gracilis Z
-
-
-
Purification (Commentary)
Commentary
Organism
CM-Sepharose column chromatography, ammonium sulfate precipitation, butyl Toyopearl column chromatography, and HiTrap Blue column chromatography
Euglena gracilis
Source Tissue
Source Tissue
Commentary
Organism
Textmining
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.00422
-
crude extract, at pH 7.2 and at 25°C
Euglena gracilis
0.671
-
after 159fold purification, at pH 7.2 and at 25°C
Euglena gracilis
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
D-galactose + NADPH + H+
14.9% activity compared to D-glucuronate
724580
Euglena gracilis
?
-
-
-
ir
D-galactose + NADPH + H+
14.9% activity compared to D-glucuronate
724580
Euglena gracilis Z
?
-
-
-
ir
D-galacturonate + NADH + H+
17.7% activity compared to NADPH
724580
Euglena gracilis
L-galactonate + NAD+
-
-
-
ir
D-galacturonate + NADPH + H+
-
724580
Euglena gracilis
L-galactonate + NADP+
-
-
-
ir
D-galacturonate + NADPH + H+
100% activity
724580
Euglena gracilis
L-galactonate + NADP+
-
-
-
ir
D-galacturonate + NADPH + H+
-
724580
Euglena gracilis Z
L-galactonate + NADP+
-
-
-
ir
D-glucose + NADPH + H+
14.6% activity compared to D-glucuronate
724580
Euglena gracilis
?
-
-
-
ir
D-glucose + NADPH + H+
14.6% activity compared to D-glucuronate
724580
Euglena gracilis Z
?
-
-
-
ir
D-glucuronate + NADPH + H+
82.4% activity compared to D-glucuronate
724580
Euglena gracilis
?
-
-
-
?
D-glucuronate + NADPH + H+
82.4% activity compared to D-glucuronate
724580
Euglena gracilis Z
?
-
-
-
?
D-xylose + NADPH + H+
29.9% activity compared to D-glucuronate
724580
Euglena gracilis
?
-
-
-
?
DL-glyceraldehyde + NADPH + H+
14.2% activity compared to D-glucuronate
724580
Euglena gracilis
?
-
-
-
ir
L-arabinose + NADPH + H+
14.2% activity compared to D-glucuronate
724580
Euglena gracilis
?
-
-
-
ir
L-galactose + NADPH + H+
18.8% activity compared to D-glucuronate
724580
Euglena gracilis
?
-
-
-
ir
additional information
the enzyme does not catalyze the reverse reaction with L-galactonate and NADP+ and does not utilize 4-nitrobenzaldehyde or menadione
724580
Euglena gracilis
?
-
-
-
-
additional information
the enzyme does not catalyze the reverse reaction with L-galactonate and NADP+ and does not utilize 4-nitrobenzaldehyde or menadione
724580
Euglena gracilis Z
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
monomer
1 * 38000, SDS-PAGE
Euglena gracilis
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.06
-
D-xylose
in 50 mM Tris-HCl (pH 7.2), at 25°C
Euglena gracilis
0.19
-
D-galacturonate
in 50 mM Tris-HCl (pH 7.2), at 25°C
Euglena gracilis
0.19
-
D-glucuronate
in 50 mM Tris-HCl (pH 7.2), at 25°C
Euglena gracilis
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.2
-
-
Euglena gracilis
pH Range
pH Minimum
pH Maximum
Commentary
Organism
6.5
9
about 75, 65 and 18.5% of maximum activity at pH 6.5, 8.0 and 9.0, respectively
Euglena gracilis
Cofactor
Cofactor
Commentary
Organism
Structure
NADPH
the enzyme prefers NADPH over NADH (ratio of 10 to 1.8)
Euglena gracilis
Activating Compound (protein specific)
Activating Compound
Commentary
Organism
Structure
H2O2
the enzyme is activated by 0.1 mM H2O2
Euglena gracilis
additional information
not influenced by EDTA, BCS, dithiothreiol, , and p-chloromercuribenzoate, L-GalL, and AsA
Euglena gracilis
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADPH
the enzyme prefers NADPH over NADH (ratio of 10 to 1.8)
Euglena gracilis
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
N-ethylmaleimide
84% residual activity at 1 mM
Euglena gracilis
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0625
-
NADPH
in 50 mM Tris-HCl (pH 7.2), at 25°C
Euglena gracilis
3.79
-
D-galacturonate
in 50 mM Tris-HCl (pH 7.2), at 25°C
Euglena gracilis
4.67
-
D-glucuronate
in 50 mM Tris-HCl (pH 7.2), at 25°C
Euglena gracilis
8.48
-
D-xylose
in 50 mM Tris-HCl (pH 7.2), at 25°C
Euglena gracilis
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Cu2+
135.3% activity at 5 mM
Euglena gracilis
additional information
not influenced by Ca2+, Co2+, Mn2+, Ni2+, Zn2+, and Fe2+
Euglena gracilis
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
38000
-
1 * 38000, SDS-PAGE
Euglena gracilis
39000
-
gel filtration
Euglena gracilis
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
D-galacturonate + NADPH + H+
Euglena gracilis
-
L-galactonate + NADP+
-
-
ir
D-galacturonate + NADPH + H+
Euglena gracilis Z
-
L-galactonate + NADP+
-
-
ir
Purification (Commentary) (protein specific)
Commentary
Organism
CM-Sepharose column chromatography, ammonium sulfate precipitation, butyl Toyopearl column chromatography, and HiTrap Blue column chromatography
Euglena gracilis
Source Tissue (protein specific)
Source Tissue
Commentary
Organism
Textmining
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.00422
-
crude extract, at pH 7.2 and at 25°C
Euglena gracilis
0.671
-
after 159fold purification, at pH 7.2 and at 25°C
Euglena gracilis
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
D-galactose + NADPH + H+
14.9% activity compared to D-glucuronate
724580
Euglena gracilis
?
-
-
-
ir
D-galactose + NADPH + H+
14.9% activity compared to D-glucuronate
724580
Euglena gracilis Z
?
-
-
-
ir
D-galacturonate + NADH + H+
17.7% activity compared to NADPH
724580
Euglena gracilis
L-galactonate + NAD+
-
-
-
ir
D-galacturonate + NADPH + H+
-
724580
Euglena gracilis
L-galactonate + NADP+
-
-
-
ir
D-galacturonate + NADPH + H+
100% activity
724580
Euglena gracilis
L-galactonate + NADP+
-
-
-
ir
D-galacturonate + NADPH + H+
-
724580
Euglena gracilis Z
L-galactonate + NADP+
-
-
-
ir
D-glucose + NADPH + H+
14.6% activity compared to D-glucuronate
724580
Euglena gracilis
?
-
-
-
ir
D-glucose + NADPH + H+
14.6% activity compared to D-glucuronate
724580
Euglena gracilis Z
?
-
-
-
ir
D-glucuronate + NADPH + H+
82.4% activity compared to D-glucuronate
724580
Euglena gracilis
?
-
-
-
?
D-glucuronate + NADPH + H+
82.4% activity compared to D-glucuronate
724580
Euglena gracilis Z
?
-
-
-
?
D-xylose + NADPH + H+
29.9% activity compared to D-glucuronate
724580
Euglena gracilis
?
-
-
-
?
DL-glyceraldehyde + NADPH + H+
14.2% activity compared to D-glucuronate
724580
Euglena gracilis
?
-
-
-
ir
L-arabinose + NADPH + H+
14.2% activity compared to D-glucuronate
724580
Euglena gracilis
?
-
-
-
ir
L-galactose + NADPH + H+
18.8% activity compared to D-glucuronate
724580
Euglena gracilis
?
-
-
-
ir
additional information
the enzyme does not catalyze the reverse reaction with L-galactonate and NADP+ and does not utilize 4-nitrobenzaldehyde or menadione
724580
Euglena gracilis
?
-
-
-
-
additional information
the enzyme does not catalyze the reverse reaction with L-galactonate and NADP+ and does not utilize 4-nitrobenzaldehyde or menadione
724580
Euglena gracilis Z
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
monomer
1 * 38000, SDS-PAGE
Euglena gracilis
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.06
-
D-xylose
in 50 mM Tris-HCl (pH 7.2), at 25°C
Euglena gracilis
0.19
-
D-galacturonate
in 50 mM Tris-HCl (pH 7.2), at 25°C
Euglena gracilis
0.19
-
D-glucuronate
in 50 mM Tris-HCl (pH 7.2), at 25°C
Euglena gracilis
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.2
-
-
Euglena gracilis
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
6.5
9
about 75, 65 and 18.5% of maximum activity at pH 6.5, 8.0 and 9.0, respectively
Euglena gracilis
Expression
Organism
Commentary
Expression
Euglena gracilis
the enzyme is not regulated by L-galactono-1,4-lactone and ascorbate
additional information
General Information
General Information
Commentary
Organism
metabolism
D-galacturonic acid reductase is a key enzyme of the ascorbate biosynthesis pathway
Euglena gracilis
General Information (protein specific)
General Information
Commentary
Organism
metabolism
D-galacturonic acid reductase is a key enzyme of the ascorbate biosynthesis pathway
Euglena gracilis
Expression (protein specific)
Organism
Commentary
Expression
Euglena gracilis
the enzyme is not regulated by L-galactono-1,4-lactone and ascorbate
additional information
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.00697
-
D-xylose
in 50 mM Tris-HCl (pH 7.2), at 25°C
Euglena gracilis
0.04135
-
D-glucuronate
in 50 mM Tris-HCl (pH 7.2), at 25°C
Euglena gracilis
0.05095
-
D-galacturonate
in 50 mM Tris-HCl (pH 7.2), at 25°C
Euglena gracilis
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.00697
-
D-xylose
in 50 mM Tris-HCl (pH 7.2), at 25°C
Euglena gracilis
0.04135
-
D-glucuronate
in 50 mM Tris-HCl (pH 7.2), at 25°C
Euglena gracilis
0.05095
-
D-galacturonate
in 50 mM Tris-HCl (pH 7.2), at 25°C
Euglena gracilis
Other publictions for EC 1.1.1.365
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)
741550
Li
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1
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-
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-
1
-
1
-
-
-
-
-
2
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1
-
1
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1
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2
-
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1
2
-
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1
-
-
-
-
-
2
-
-
1
-
1
-
-
-
1
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-
-
-
2
2
-
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-
742938
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Naganishia diffluens, Naganishia diffluens OPU-FC11
J. Biosci. Bioeng.
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-
-
1
-
1
-
-
-
-
-
-
2
-
5
-
-
-
-
-
-
-
-
4
1
-
-
-
-
1
-
-
2
-
-
-
-
-
1
2
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
4
1
-
-
-
-
1
-
-
-
-
1
1
-
-
-
743438
Lim
-
Transgenic tomato plants expr ...
Fragaria x ananassa
Plant Biotechnol. Rep.
10
105-116
2016
-
-
1
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
2
-
-
-
-
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-
-
-
-
-
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-
-
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-
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1
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1
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-
-
-
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-
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-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
742163
Amaya
Increased antioxidant capacit ...
Fragaria x ananassa
Biotechnol. J.
10
490-500
2015
-
-
1
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
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-
-
-
-
-
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-
-
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
743465
Cai
-
Ectopic expression of FaGalUR ...
Fragaria x ananassa
Plant Growth Regul.
76
187-197
2015
-
1
1
-
1
-
-
-
-
-
-
1
-
1
-
-
-
-
-
2
-
-
1
-
1
-
-
-
1
-
-
2
-
-
-
-
1
1
2
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
2
-
-
1
-
1
-
-
-
1
-
-
-
-
1
1
-
-
-
741576
Sloothaak
Overexpression of the Aspergi ...
Aspergillus niger, Aspergillus niger CBS 120.49
AMB Express
4
66
2014
-
-
1
-
-
-
-
-
-
-
-
2
-
3
-
-
-
-
-
1
-
-
2
-
-
-
-
-
1
-
-
2
-
-
-
-
-
1
2
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
1
-
-
2
-
-
-
-
-
1
-
-
-
-
1
1
-
-
-
741553
Yang
-
Accumulation of vitamin C and ...
Fragaria x ananassa
Adv. Mater. Res.
749
277-282
2013
-
-
1
-
1
-
-
-
-
-
-
1
-
1
-
-
-
-
-
2
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
1
2
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
2
-
-
1
-
-
-
-
-
-
-
-
-
-
2
2
-
-
-
721358
Kuivanen
Engineering filamentous fungi ...
Aspergillus niger, Aspergillus niger ATCC 1015, Trichoderma reesei
Appl. Environ. Microbiol.
78
8676-8683
2012
-
-
-
-
-
-
-
-
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3
-
9
-
-
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-
-
-
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3
-
-
-
-
-
-
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2
-
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2
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3
-
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3
-
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-
-
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-
725116
Venkatesh
-
Chlorophyll a fluorescence tra ...
Fragaria x ananassa
Hortic. Environ. Biotechnol.
53
320-328
2012
-
-
1
-
-
-
-
-
-
-
-
-
-
1
-
-
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-
-
-
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