BRENDA - Enzyme Database show
show all sequences of 1.14.11.15

Partial purification and characterization of gibberellin 3beta-hydroxylase from immature seeds of Phaseolus vulgaris

Kwak, S.S.; Kamiya, Y.; Sakurai, A.; Takahashi, N.; Graebe, J.; Plant Cell Physiol. 29, 935-943 (1988)
No PubMed abstract available

Data extracted from this reference:

Activating Compound
Activating Compound
Commentary
Organism
Structure
bovine serum albumin
2 mg/ml, 30% activation of purified enzyme, stimulation by bovine serum albumin and catalase is additive and less pronounced in crude cell extract
Phaseolus vulgaris
catalase
0.5 mg/ml, 30% activation of purified enzyme, stimulation by catalase and bovine serum albumin is additive and less pronounced in crude cell extract
Phaseolus vulgaris
Inhibitors
Inhibitors
Commentary
Organism
Structure
2,2'-bipyridyl
-
Phaseolus vulgaris
Cd2+
0.5 mM, strong
Phaseolus vulgaris
Co2+
0.5 mM, strong
Phaseolus vulgaris
Cu2+
0.5 mM, strong
Phaseolus vulgaris
gibberellin a15
inhibits 3beta-hydroxylation of gibberellin a20
Phaseolus vulgaris
gibberellin a20
-
Phaseolus vulgaris
gibberellin a44
inhibits 3beta-hydroxylation of gibberellin a20
Phaseolus vulgaris
gibberellin a5
inhibits 3beta-hydroxylation of gibberellin a20
Phaseolus vulgaris
gibberellin a9
inhibits 3beta-hydroxylation of gibberellin a20
Phaseolus vulgaris
Hg2+
0.5 mM, strong
Phaseolus vulgaris
Mn2+
0.5 mM, strong
Phaseolus vulgaris
Ni2+
0.5 mM, strong
Phaseolus vulgaris
Zn2+
0.5 mM, strong
Phaseolus vulgaris
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.00029
-
gibberellin a20
-
Phaseolus vulgaris
0.00033
-
gibberellin a9
-
Phaseolus vulgaris
0.25
-
2-oxoglutarate
with gibberellin a20 as cosubstrate
Phaseolus vulgaris
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Fe2+
activates, at least 0.2 mM for full activation
Phaseolus vulgaris
Fe3+
activation in the presence of ascorbate
Phaseolus vulgaris
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
42000
-
gel filtration
Phaseolus vulgaris
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Phaseolus vulgaris
-
-
-
Purification (Commentary)
Commentary
Organism
partial
Phaseolus vulgaris
Source Tissue
Source Tissue
Commentary
Organism
Textmining
seed
immature
Phaseolus vulgaris
-
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
additional information
-
-
Phaseolus vulgaris
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
gibberellin a20 + 2-oxoglutarate + O2
-
439262
Phaseolus vulgaris
gibberellin a1 + succinate + CO2
-
439262
Phaseolus vulgaris
?
gibberellin a9 + 2-oxoglutarate + O2
-
439262
Phaseolus vulgaris
gibberellin a4 + succinate + CO2
-
-
-
?
Subunits
Subunits
Commentary
Organism
monomer
1 * 42000, SDS-PAGE
Phaseolus vulgaris
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.7
-
-
Phaseolus vulgaris
pH Range
pH Minimum
pH Maximum
Commentary
Organism
7
9
about half-maximal activity at pH 7.0 and pH 9.0
Phaseolus vulgaris
Cofactor
Cofactor
Commentary
Organism
Structure
ascorbate
5 mM, 10fold stimulation
Phaseolus vulgaris
Activating Compound (protein specific)
Activating Compound
Commentary
Organism
Structure
bovine serum albumin
2 mg/ml, 30% activation of purified enzyme, stimulation by bovine serum albumin and catalase is additive and less pronounced in crude cell extract
Phaseolus vulgaris
catalase
0.5 mg/ml, 30% activation of purified enzyme, stimulation by catalase and bovine serum albumin is additive and less pronounced in crude cell extract
Phaseolus vulgaris
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
ascorbate
5 mM, 10fold stimulation
Phaseolus vulgaris
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
2,2'-bipyridyl
-
Phaseolus vulgaris
Cd2+
0.5 mM, strong
Phaseolus vulgaris
Co2+
0.5 mM, strong
Phaseolus vulgaris
Cu2+
0.5 mM, strong
Phaseolus vulgaris
gibberellin a15
inhibits 3beta-hydroxylation of gibberellin a20
Phaseolus vulgaris
gibberellin a20
-
Phaseolus vulgaris
gibberellin a44
inhibits 3beta-hydroxylation of gibberellin a20
Phaseolus vulgaris
gibberellin a5
inhibits 3beta-hydroxylation of gibberellin a20
Phaseolus vulgaris
gibberellin a9
inhibits 3beta-hydroxylation of gibberellin a20
Phaseolus vulgaris
Hg2+
0.5 mM, strong
Phaseolus vulgaris
Mn2+
0.5 mM, strong
Phaseolus vulgaris
Ni2+
0.5 mM, strong
Phaseolus vulgaris
Zn2+
0.5 mM, strong
Phaseolus vulgaris
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.00029
-
gibberellin a20
-
Phaseolus vulgaris
0.00033
-
gibberellin a9
-
Phaseolus vulgaris
0.25
-
2-oxoglutarate
with gibberellin a20 as cosubstrate
Phaseolus vulgaris
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Fe2+
activates, at least 0.2 mM for full activation
Phaseolus vulgaris
Fe3+
activation in the presence of ascorbate
Phaseolus vulgaris
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
42000
-
gel filtration
Phaseolus vulgaris
Purification (Commentary) (protein specific)
Commentary
Organism
partial
Phaseolus vulgaris
Source Tissue (protein specific)
Source Tissue
Commentary
Organism
Textmining
seed
immature
Phaseolus vulgaris
-
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
additional information
-
-
Phaseolus vulgaris
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
gibberellin a20 + 2-oxoglutarate + O2
-
439262
Phaseolus vulgaris
gibberellin a1 + succinate + CO2
-
439262
Phaseolus vulgaris
?
gibberellin a9 + 2-oxoglutarate + O2
-
439262
Phaseolus vulgaris
gibberellin a4 + succinate + CO2
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
monomer
1 * 42000, SDS-PAGE
Phaseolus vulgaris
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.7
-
-
Phaseolus vulgaris
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
7
9
about half-maximal activity at pH 7.0 and pH 9.0
Phaseolus vulgaris
Other publictions for EC 1.14.11.15
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)
741490
Nett
Characterization of CYP115 as ...
Mesorhizobium loti, Mesorhizobium loti CJ3sym, Mesorhizobium loti MAFF303099, Mesorhizobium loti NZP2014, Mesorhizobium loti NZP2042, Mesorhizobium loti R7A, Mesorhizobium loti R88b, Mesorhizobium loti Tono, Mesorhizobium loti USDA3471, Mesorhizobium sp., Mesorhizobium sp. AA22, Mesorhizobium sp. SEMIA 3007, Mesorhizobium sp. STM4661, Mesorhizobium sp. WSM2561, Mesorhizobium sp. WSM3626, Rhizobium favelukesii, Rhizobium favelukesii OR191, Rhizobium mongolense, Rhizobium mongolense USDA1844, Sinorhizobium arboris, Sinorhizobium arboris LMG 14919, Sinorhizobium fredii, Sinorhizobium fredii CCBAU83666 AJQR, Sinorhizobium medicae, Sinorhizobium medicae WSM1369, Sinorhizobium meliloti, Sinorhizobium meliloti WSM4191, Sinorhizobium terangae, Sinorhizobium terangae WSM1721
ACS Chem. Biol.
12
912-917
2017
-
-
2
-
-
-
-
-
-
-
-
33
-
45
-
-
-
-
-
-
-
-
66
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
33
-
-
-
-
-
-
-
-
66
-
-
-
-
-
-
-
-
-
-
10
14
-
-
-
741544
Otani
-
Overexpression of the gibbere ...
Torenia fournieri
Acta Hortic.
1025
23-30
2014
-
-
1
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
743493
Chen
The maize DWARF1 encodes a gi ...
Zea mays
Plant Physiol.
166
2028-2039
2014
-
-
1
-
-
-
-
-
4
-
-
3
-
2
-
-
-
-
-
2
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
4
-
-
3
-
-
-
-
-
2
-
-
3
-
-
-
-
-
-
-
-
-
1
2
2
1
-
-
725686
Kim
Functional mechanism of plant ...
Arabidopsis thaliana, Arabidopsis thaliana Col-0, Capsicum annuum
J. Microbiol. Biotechnol.
22
1375-1380
2012
2
-
1
-
-
-
8
-
-
2
-
2
-
7
-
-
1
-
-
3
-
-
4
-
2
-
-
-
2
-
-
-
-
-
-
2
-
1
-
-
-
-
-
8
-
-
-
2
-
2
-
-
-
1
-
3
-
-
4
-
2
-
-
-
2
-
-
-
-
3
3
-
-
-
689507
Gallego-Giraldo
Gibberellin homeostasis in tob ...
Nicotiana tabacum
Plant Cell Physiol.
49
679-690
2008
-
-
1
-
1
-
1
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
689516
Dalmadi
Dwarf plants of diploid Medica ...
Medicago sativa
Plant Cell Rep.
27
1271-1279
2008
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
700749
Kim
A membrane-bound NAC transcrip ...
Arabidopsis thaliana
Plant J.
55
77-88
2008
-
-
1
-
1
-
-
-
-
1
-
2
-
2
-
-
1
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
1
-
2
-
-
-
1
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
700861
Urakami
Immunomodulation of gibberelli ...
Arabidopsis thaliana
Planta
228
863-873
2008
-
-
1
-
1
-
-
-
-
-
-
-
-
2
-
-
1
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
1
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
689465
Hirano
The GID1-mediated gibberellin ...
Selaginella moellendorffii
Plant Cell
19
3058-3079
2007
-
-
-
-
-
-
1
-
-
-
-
1
-
8
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
689498
Gallego-Giraldo
Flowering in tobacco needs gib ...
Nicotiana tabacum
Plant Cell Physiol.
48
615-625
2007
-
-
-
-
-
-
-
-
-
-
-
1
-
3
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
676505
Mitchum
Distinct and overlapping roles ...
Arabidopsis thaliana
Plant J.
45
804-818
2006
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
2
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
676607
Benschop
Long-term submergence-induced ...
Rumex palustris
Plant Physiol.
141
1644-1652
2006
-
-
-
-
-
-
-
-
-
-
-
-
-
3
-
-
1
-
-
2
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
2
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
676701
Appleford
Function and transcript analys ...
Triticum aestivum
Planta
223
568-582
2006
-
-
1
-
-
-
-
2
-
-
-
-
-
3
-
-
1
-
-
2
-
-
6
-
-
-
-
-
-
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
6
-
-
-
-
-
-
-
3
-
6
-
-
6
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
672812
Lester
Purification and kinetic studi ...
Arabidopsis thaliana
BMC Plant Biol.
5
19
2005
-
-
1
-
-
-
1
1
-
-
1
-
-
5
-
-
1
-
-
-
2
-
1
-
-
-
-
-
-
-
1
-
-
-
-
-
-
1
-
-
-
-
-
1
-
1
-
-
1
-
-
-
-
1
-
-
2
-
1
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
676398
Suzuki
-
Specific expression of the gib ...
Hordeum vulgare
Plant Biotechnol.
22
195-200
2005
-
-
1
-
-
-
1
-
-
-
-
-
-
1
-
-
-
-
-
2
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
2
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
676453
Solfanelli
Expression of two genes encodi ...
Phaseolus coccineus
Plant Cell Physiol.
46
1116-1124
2005
-
-
1
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
2
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
660074
King
Gibberellin structure and func ...
Pisum sativum
Physiol. Plant.
120
287-297
2004
-
-
-
-
-
-
10
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
10
-
-
-
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
660239
Zhou
16,17-dihydro gibberellin A5 c ...
Arabidopsis sp.
Plant Physiol.
135
1000-1007
2004
-
-
1
-
-
-
3
2
-
-
-
-
-
3
-
-
-
-
-
-
-
-
3
-
-
-
-
-
1
1
-
-
-
-
-
-
-
1
-
-
-
-
-
3
-
2
-
-
-
-
-
-
-
-
-
-
-
-
3
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
660243
Israelsson
Cloning and overproduction of ...
Populus tremula x Populus tremuloides
Plant Physiol.
135
221-230
2004
-
-
1
-
-
-
-
-
-
-
-
2
-
4
-
-
-
-
-
3
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
3
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
660218
Ozga
Pollination-, development-, an ...
Pisum sativum
Plant Physiol.
131
1137-1146
2003
-
-
-
-
-
-
-
-
-
-
-
1
-
2
-
-
-
-
-
6
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
6
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
439267
Itoh
Cloning and functional analysi ...
Oryza sativa
Proc. Natl. Acad. Sci. USA
98
8909-8914
2001
-
-
1
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
439264
Ait-Ali
Regulation of gibberellin 20-o ...
Pisum sativum
Plant Physiol.
121
783-791
1999
-
-
-
-
-
-
-
-
-
-
-
1
-
2
-
-
-
-
-
2
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
2
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
439265
Yamaguchi
Phytochrome regulation and dif ...
Arabidopsis thaliana
Plant Cell
10
2115-2126
1998
-
-
-
-
-
-
-
-
-
-
-
1
-
2
-
-
-
-
-
2
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
2
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
439266
Lange
-
Separation and characterizatio ...
Cucurbita maxima
Planta
195
98-107
1994
2
-
-
-
-
-
-
1
-
1
1
1
-
1
-
-
1
-
-
2
1
-
3
1
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
1
-
1
1
1
-
-
-
1
-
2
1
-
3
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
439263
Saito
Effects of 3-Methylgibberellin ...
Cucurbita maxima, Phaseolus vulgaris
Biosci. Biotechnol. Biochem.
56
1046-1052
1992
-
-
-
-
-
-
6
-
2
-
-
2
-
2
-
-
-
-
-
2
-
-
6
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
6
-
-
2
-
-
2
-
-
-
-
-
2
-
-
6
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
439261
Saito
-
Effects of deoxygibberellin C ...
Cucurbita maxima, Phaseolus vulgaris
Plant Cell Physiol.
32
239-245
1991
-
-
-
-
-
-
4
-
-
-
-
-
-
2
-
-
-
-
-
2
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
2
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
439262
Kwak
-
Partial purification and chara ...
Phaseolus vulgaris
Plant Cell Physiol.
29
935-943
1988
2
-
-
-
-
-
13
3
-
2
1
-
-
1
-
-
1
-
-
1
1
-
2
1
-
-
-
-
1
1
-
1
-
-
-
2
-
-
1
-
-
-
-
13
-
3
-
2
1
-
-
-
-
1
-
1
1
-
2
1
-
-
-
-
1
1
-
-
-
-
-
-
-
-