BRENDA - Enzyme Database show

Production of 10S-hydroxy-8(E)-octadecenoic acid from oleic acid by whole recombinant Escherichia coli cells expressing 10S-dioxygenase from Nostoc punctiforme PCC 73102 with the aid of a chaperone

Kim, M.J.; Seo, M.J.; Shin, K.C.; Oh, D.K.; Biotechnol. Lett. 39, 133-139 (2017)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
1.13.11.77
chaperone Gro7
activates the enzyme about 200%
Nostoc punctiforme
Cloned(Commentary)
EC Number
Commentary
Organism
1.13.11.77
recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3), coexpression with chaperone Gro7
Nostoc punctiforme
Engineering
EC Number
Amino acid exchange
Commentary
Organism
1.13.11.77
additional information
optimization of conditions for 10S-hydroxy-8(E)-octadecenoic acid production from oleic acid by recombinant Escherichia coli cells expressing Nostoc punctiforme 10S-dioxygenase with the aid of a chaperone plasmid. Optimal conditions are pH 9.0, 35°C, 15 % v/v dimethyl sulfoxide, 40 g cells/l, and 10 g oleic acid/l. The activity of recombinant cells expressing 10S-dioxygenase is increased by 200% with the aid of chaperone Gro7. Production of 10S-hydroxy-8(E)-octadecenoic acid, 10S-hydroxy-8,12(E,Z)-octadecadienoic acid, and 10S-hydroxy-8,12,15(E,Z,Z)-octadecatrienoic acid from unsaturated fatty acids including oleic acid, linoleic acid, and a-linolenic acid, respectively. The soluble 10S-dioxygenase content in the crude extract of recombinant cells co-expressing the 10S-dioxygenase gene with pGro7 is higher than that of recombinant cells co-expressing pKJE7 or pG-KJE8. Production of 360 mg/g/h 10S-hydroxy-8(E)-octadecenoic acid from oleic acid
Nostoc punctiforme
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.13.11.77
0.096
-
alpha-linolenate
recombinant enzyme, pH 9.0, 35°C
Nostoc punctiforme
1.13.11.77
0.236
-
linoleate
recombinant enzyme, pH 9.0, 35°C
Nostoc punctiforme
1.13.11.77
0.345
-
oleate
recombinant enzyme, pH 9.0, 35°C
Nostoc punctiforme
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.13.11.77
alpha-linolenate + O2
Nostoc punctiforme
-
(8E,10S,12Z,15Z)-10-hydroperoxyoctadeca-8,12,15-trienoate
-
-
?
1.13.11.77
linoleate + O2
Nostoc punctiforme
-
(8E,10S,12Z)-10-hydroperoxyoctadeca-8,12-dienoate
-
-
?
1.13.11.77
oleate + O2
Nostoc punctiforme
preferred substrate
(8E,10S)-10-hydroperoxyoctadeca-8-enoate
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.13.11.77
Nostoc punctiforme
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.13.11.77
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and dialysis
Nostoc punctiforme
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.13.11.77
0.096
-
purified recombinant enzyme, substrate alpha-linolenate, pH 9.0, 35°C
Nostoc punctiforme
1.13.11.77
0.236
-
purified recombinant enzyme, substrate linoleate, pH 9.0, 35°C
Nostoc punctiforme
1.13.11.77
0.345
-
purified recombinant enzyme, substrate oleate, pH 9.0, 35°C
Nostoc punctiforme
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.13.11.77
alpha-linolenate + O2
-
744582
Nostoc punctiforme
(8E,10S,12Z,15Z)-10-hydroperoxyoctadeca-8,12,15-trienoate
-
-
-
?
1.13.11.77
linoleate + O2
-
744582
Nostoc punctiforme
(8E,10S,12Z)-10-hydroperoxyoctadeca-8,12-dienoate
-
-
-
?
1.13.11.77
additional information
optimization of assay conditions, overview
744582
Nostoc punctiforme
?
-
-
-
-
1.13.11.77
oleate + O2
preferred substrate
744582
Nostoc punctiforme
(8E,10S)-10-hydroperoxyoctadeca-8-enoate
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.13.11.77
?
x * 63000, recombinant His-tagged enzyme, SDS-PAGE
Nostoc punctiforme
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.13.11.77
35
-
recombinant His-tagged enzyme
Nostoc punctiforme
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.13.11.77
9
-
recombinant His-tagged enzyme
Nostoc punctiforme
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
1.13.11.77
chaperone Gro7
activates the enzyme about 200%
Nostoc punctiforme
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.13.11.77
recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3), coexpression with chaperone Gro7
Nostoc punctiforme
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.13.11.77
additional information
optimization of conditions for 10S-hydroxy-8(E)-octadecenoic acid production from oleic acid by recombinant Escherichia coli cells expressing Nostoc punctiforme 10S-dioxygenase with the aid of a chaperone plasmid. Optimal conditions are pH 9.0, 35°C, 15 % v/v dimethyl sulfoxide, 40 g cells/l, and 10 g oleic acid/l. The activity of recombinant cells expressing 10S-dioxygenase is increased by 200% with the aid of chaperone Gro7. Production of 10S-hydroxy-8(E)-octadecenoic acid, 10S-hydroxy-8,12(E,Z)-octadecadienoic acid, and 10S-hydroxy-8,12,15(E,Z,Z)-octadecatrienoic acid from unsaturated fatty acids including oleic acid, linoleic acid, and a-linolenic acid, respectively. The soluble 10S-dioxygenase content in the crude extract of recombinant cells co-expressing the 10S-dioxygenase gene with pGro7 is higher than that of recombinant cells co-expressing pKJE7 or pG-KJE8. Production of 360 mg/g/h 10S-hydroxy-8(E)-octadecenoic acid from oleic acid
Nostoc punctiforme
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.13.11.77
0.096
-
alpha-linolenate
recombinant enzyme, pH 9.0, 35°C
Nostoc punctiforme
1.13.11.77
0.236
-
linoleate
recombinant enzyme, pH 9.0, 35°C
Nostoc punctiforme
1.13.11.77
0.345
-
oleate
recombinant enzyme, pH 9.0, 35°C
Nostoc punctiforme
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.13.11.77
alpha-linolenate + O2
Nostoc punctiforme
-
(8E,10S,12Z,15Z)-10-hydroperoxyoctadeca-8,12,15-trienoate
-
-
?
1.13.11.77
linoleate + O2
Nostoc punctiforme
-
(8E,10S,12Z)-10-hydroperoxyoctadeca-8,12-dienoate
-
-
?
1.13.11.77
oleate + O2
Nostoc punctiforme
preferred substrate
(8E,10S)-10-hydroperoxyoctadeca-8-enoate
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.13.11.77
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and dialysis
Nostoc punctiforme
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.13.11.77
0.096
-
purified recombinant enzyme, substrate alpha-linolenate, pH 9.0, 35°C
Nostoc punctiforme
1.13.11.77
0.236
-
purified recombinant enzyme, substrate linoleate, pH 9.0, 35°C
Nostoc punctiforme
1.13.11.77
0.345
-
purified recombinant enzyme, substrate oleate, pH 9.0, 35°C
Nostoc punctiforme
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.13.11.77
alpha-linolenate + O2
-
744582
Nostoc punctiforme
(8E,10S,12Z,15Z)-10-hydroperoxyoctadeca-8,12,15-trienoate
-
-
-
?
1.13.11.77
linoleate + O2
-
744582
Nostoc punctiforme
(8E,10S,12Z)-10-hydroperoxyoctadeca-8,12-dienoate
-
-
-
?
1.13.11.77
additional information
optimization of assay conditions, overview
744582
Nostoc punctiforme
?
-
-
-
-
1.13.11.77
oleate + O2
preferred substrate
744582
Nostoc punctiforme
(8E,10S)-10-hydroperoxyoctadeca-8-enoate
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.13.11.77
?
x * 63000, recombinant His-tagged enzyme, SDS-PAGE
Nostoc punctiforme
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.13.11.77
35
-
recombinant His-tagged enzyme
Nostoc punctiforme
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.13.11.77
9
-
recombinant His-tagged enzyme
Nostoc punctiforme