additional information
reaction conditions for the production of (7S,8S,9Z,12Z)-dihydroxyoctadeca-9,12-dienoic acid from linoleic acid by recombinant Escherichia coli expressing 7,8-linoleate diol synthase from Glomerella cingulata are optimized using response surface methodology, overview. The optimal reaction conditions are pH 7.0, 18.6°C, 10.8% v/v dimethyl sulfoxide, 44.9 g/l cells, and 14.3 g/l linoleic acid, with agitation at 256 rpm. Under these conditions, recombinant cells produce 7,8-dihydroxy unsaturated fatty acids in the range of 7.0-9.8 g/l from 14.3 g/l linoleic acid, 14.3 g/l oleic acid, and plant oil hydrolysates such as waste oil and olive oil containing 14.3 g/l linoleic acid or oleic acid. Comparisons of quantitative production of dihydroxy unsaturated fatty acids by microorganisms
Colletotrichum gloeosporioides
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
742730
Colletotrichum gloeosporioides
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
r
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
isomerase reaction of the C-terminal domain
742730
Colletotrichum gloeosporioides
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
r
(9Z)-8-hydroperoxyoctadeca-9-enoic acid
isomerase reaction of the C-terminal domain
742730
Colletotrichum gloeosporioides
(9Z)-7,8-dihydroxyoctadeca-9-enoic acid
r
(9Z,12Z,15Z)-8-hydroperoxyoctadeca-9,12,15-trienoic acid
isomerase reaction of the C-terminal domain
742730
Colletotrichum gloeosporioides
(9Z,12Z,15Z)-7,8-dihydroxyoctadeca-9,12,15-trienoic acid
r
alpha-linolenate + O2
via (9Z,12Z,15Z)-8-hydroperoxyoctadeca-9,12,15-trienoic acid, whole enzyme reaction
742730
Colletotrichum gloeosporioides
(9Z,12Z,15Z)-7,8-dihydroperoxyoctadeca-9,12,15-trienoic acid
?
linoleate
via (8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate, whole enzyme reaction, best substrate
742730
Colletotrichum gloeosporioides
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
r
additional information
the bifunctional enzyme converts linoleic acid to a product, identified as (9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoic acid (7,8-DiHODE) by liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) and nuclear magnetic resonance (NMR) spectroscopy. The specific activity and catalytic efficiency (kcat/Km) of 7,8-diol synthase for the conversion of fatty acid to dihydroxy fatty acid follows the descending order linoleic acid, alpha-linolenic acid, and oleic acid, indicating that the enzyme is a 7,8-linoleate diol synthase (7,8-LDS). The reaction via (9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoic acid (8-HPODE) as an intermediate and the accumulation of 8-HPODE is due to a higher 8-dioxygenase activity in the N-terminal domain than hydroperoxide isomerase activity in the C-terminal domain, EC 5.4.4.6. 8-HPODE is subsequently isomerized to (7S,8S,9Z,12Z)-dihydroxyoctadeca-9,12-dienoic acid (7,8-DiHODE) or (5S,8R,9Z,12Z)-dihydroxyoctadeca-9,12-dienoic acid (5,8-DiHODE) by the C-terminal hydroperoxide isomerase domain of diol synthase
742730
Colletotrichum gloeosporioides
?
oleate + O2
via (9Z)-8-hydroperoxyoctadeca-9-enoic acid, whole enzyme reaction
742730
Colletotrichum gloeosporioides
(9Z)-7,8-dihydroxyoctadeca-9-enoic acid
?
additional information
reaction conditions for the production of (7S,8S,9Z,12Z)-dihydroxyoctadeca-9,12-dienoic acid from linoleic acid by recombinant Escherichia coli expressing 7,8-linoleate diol synthase from Glomerella cingulata are optimized using response surface methodology, overview. The optimal reaction conditions are pH 7.0, 18.6°C, 10.8% v/v dimethyl sulfoxide, 44.9 g/l cells, and 14.3 g/l linoleic acid, with agitation at 256 rpm. Under these conditions, recombinant cells produce 7,8-dihydroxy unsaturated fatty acids in the range of 7.0-9.8 g/l from 14.3 g/l linoleic acid, 14.3 g/l oleic acid, and plant oil hydrolysates such as waste oil and olive oil containing 14.3 g/l linoleic acid or oleic acid. Comparisons of quantitative production of dihydroxy unsaturated fatty acids by microorganisms
Colletotrichum gloeosporioides
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
742730
Colletotrichum gloeosporioides
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
r
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
isomerase reaction of the C-terminal domain
742730
Colletotrichum gloeosporioides
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
r
(9Z)-8-hydroperoxyoctadeca-9-enoic acid
isomerase reaction of the C-terminal domain
742730
Colletotrichum gloeosporioides
(9Z)-7,8-dihydroxyoctadeca-9-enoic acid
r
(9Z,12Z,15Z)-8-hydroperoxyoctadeca-9,12,15-trienoic acid
isomerase reaction of the C-terminal domain
742730
Colletotrichum gloeosporioides
(9Z,12Z,15Z)-7,8-dihydroxyoctadeca-9,12,15-trienoic acid
r
alpha-linolenate + O2
via (9Z,12Z,15Z)-8-hydroperoxyoctadeca-9,12,15-trienoic acid, whole enzyme reaction
742730
Colletotrichum gloeosporioides
(9Z,12Z,15Z)-7,8-dihydroperoxyoctadeca-9,12,15-trienoic acid
?
linoleate
via (8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate, whole enzyme reaction, best substrate
742730
Colletotrichum gloeosporioides
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
r
additional information
the bifunctional enzyme converts linoleic acid to a product, identified as (9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoic acid (7,8-DiHODE) by liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) and nuclear magnetic resonance (NMR) spectroscopy. The specific activity and catalytic efficiency (kcat/Km) of 7,8-diol synthase for the conversion of fatty acid to dihydroxy fatty acid follows the descending order linoleic acid, alpha-linolenic acid, and oleic acid, indicating that the enzyme is a 7,8-linoleate diol synthase (7,8-LDS). The reaction via (9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoic acid (8-HPODE) as an intermediate and the accumulation of 8-HPODE is due to a higher 8-dioxygenase activity in the N-terminal domain than hydroperoxide isomerase activity in the C-terminal domain, EC 5.4.4.6. 8-HPODE is subsequently isomerized to (7S,8S,9Z,12Z)-dihydroxyoctadeca-9,12-dienoic acid (7,8-DiHODE) or (5S,8R,9Z,12Z)-dihydroxyoctadeca-9,12-dienoic acid (5,8-DiHODE) by the C-terminal hydroperoxide isomerase domain of diol synthase
742730
Colletotrichum gloeosporioides
?
oleate + O2
via (9Z)-8-hydroperoxyoctadeca-9-enoic acid, whole enzyme reaction
742730
Colletotrichum gloeosporioides
(9Z)-7,8-dihydroxyoctadeca-9-enoic acid
?
741712
Seo
Characterization of a recombi ...
Colletotrichum gloeosporioides, Colletotrichum gloeosporioides KACC 40961
Appl. Microbiol. Biotechnol.
100
3087-3099
2016
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742730
Seo
Production of 7,8-dihydroxy u ...
Colletotrichum gloeosporioides, Colletotrichum gloeosporioides KACC 40961
J. Agric. Food Chem.
64
8555-8562
2016
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702018
Garscha
Pichia expression and mutagene ...
Gaeumannomyces graminis, Gaeumannomyces graminis var. avenae
Biochem. Biophys. Res. Commun.
373
579-583
2008
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703700
Garscha
Critical amino acids for the 8 ...
Gaeumannomyces graminis
FEBS Lett.
582
3547-3551
2008
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285246
Hörnsten
Cloning of linoleate diol synt ...
Gaeumannomyces graminis
J. Biol. Chem.
274
28219-28224
1999
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1
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285251
Oliw
Catalytic properties of linole ...
Gaeumannomyces graminis
Adv. Exp. Med. Biol.
469
679-685
1999
2
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285244
Su
A protein radical and ferryl i ...
Gaeumannomyces graminis
J. Biol. Chem.
273
20744-20751
1998
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285249
Su
Studies on linoleic acid 8R-di ...
Gaeumannomyces graminis
Lipids
30
43-50
1995
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285247
Brodowsky
BW A4C and other hydroxamic ac ...
Gaeumannomyces graminis
Eur. J. Pharmacol.
254
43-47
1994
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285250
Hamberg
Sequential oxygenation of lino ...
Gaeumannomyces graminis, Gaeumannomyces graminis var.graminis
Arch. Biochem. Biophys.
309
77-80
1994
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285245
Brodowsky
A linoleic acid (8R)-dioxygena ...
Gaeumannomyces graminis
J. Biol. Chem.
267
14738-14745
1992
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