BRENDA - Reference to 5.4.4.6; Id = 742730

DMSO

activates the whole enzyme activity

Colletotrichum gloeosporioides

recombinant expression of His-tagged enzyme in Escherichia coli strain ER2566

Colletotrichum gloeosporioides

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

Colletotrichum gloeosporioides

-

(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate

-

r

Colletotrichum gloeosporioides

A0A0S2SWE4

i.e. Colletotrichum gloeosporioides

-

Colletotrichum gloeosporioides KACC 40961

A0A0S2SWE4

i.e. Colletotrichum gloeosporioides

-

recombinant His-tagged wild-enzyme from Escherichia coli strain ER2566 by nickel affinity chromatography

Colletotrichum gloeosporioides

mycelium

-

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

-

?

7,8-diol synthase

-

Colletotrichum gloeosporioides

7,8-LDS

-

Colletotrichum gloeosporioides

7,8-linoleate diol synthase

-

Colletotrichum gloeosporioides

37

-

assay at

Colletotrichum gloeosporioides

7.5

-

assay at

Colletotrichum gloeosporioides

DMSO

activates the whole enzyme activity

Colletotrichum gloeosporioides

recombinant expression of His-tagged enzyme in Escherichia coli strain ER2566

Colletotrichum gloeosporioides

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

Colletotrichum gloeosporioides

-

(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate

-

r

recombinant His-tagged wild-enzyme from Escherichia coli strain ER2566 by nickel affinity chromatography

Colletotrichum gloeosporioides

mycelium

-

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

-

?

37

-

assay at

Colletotrichum gloeosporioides

7.5

-

assay at

Colletotrichum gloeosporioides

741712

Seo

Characterization of a recombi ...

Colletotrichum gloeosporioides, Colletotrichum gloeosporioides KACC 40961

Appl. Microbiol. Biotechnol.

100

3087-3099

2016

-

1

-

11

-

4

-

1

-

3

-

1

-

1

4

-

5

2

3

1

-

4

2

-

1

-

11

-

4

-

1

-

1

-

1

4

-

5

2

1

-

4

2

-

4

742730

Seo

Production of 7,8-dihydroxy u ...

Colletotrichum gloeosporioides, Colletotrichum gloeosporioides KACC 40961

J. Agric. Food Chem.

64

8555-8562

2016

1

-

1

-

1

-

1

-

3

-

1

-

1

-

8

-

3

1

-

1

-

1

-

1

-

1

-

1

-

1

-

1

-

8

-

1

-

1

-

702018

Garscha

Pichia expression and mutagene ...

Gaeumannomyces graminis, Gaeumannomyces graminis var. avenae

Biochem. Biophys. Res. Commun.

373

579-583

2008

-

1

-

3

-

4

-

4

-

1

-

8

-

2

-

2

-

4

-

4

-

2

-

8

-

703700

Garscha

Critical amino acids for the 8 ...

Gaeumannomyces graminis

FEBS Lett.

582

3547-3551

2008

-

1

-

11

-

1

-

1

-

1

-

2

-

1

-

1

-

11

-

1

-

1

-

285246

Hörnsten

Cloning of linoleate diol synt ...

Gaeumannomyces graminis

J. Biol. Chem.

274

28219-28224

1999

-

1

-

1

-

1

-

1

-

1

-

1

-

1

-

285251

Oliw

Catalytic properties of linole ...

Gaeumannomyces graminis

Adv. Exp. Med. Biol.

469

679-685

1999

2

-

2

-

1

-

1

-

1

-

1

-

1

-

2

-

1

-

2

-

1

-

1

-

1

-

285244

Su

A protein radical and ferryl i ...

Gaeumannomyces graminis

J. Biol. Chem.

273

20744-20751

1998

-

1

-

2

-

2

-

1

-

1

-

4

-

1

-

2

-

2

-

2

-

2

-

2

-

4

-

285249

Su

Studies on linoleic acid 8R-di ...

Gaeumannomyces graminis

Lipids

30

43-50

1995

-

1

-

1

-

1

-

1

-

1

-

1

-

1

-

1

-

1

-

1

-

1

-

285247

Brodowsky

BW A4C and other hydroxamic ac ...

Gaeumannomyces graminis

Eur. J. Pharmacol.

254

43-47

1994

-

1

-

1

-

1

-

1

-

1

-

1

-

1

-

1

-

1

-

285250

Hamberg

Sequential oxygenation of lino ...

Gaeumannomyces graminis, Gaeumannomyces graminis var.graminis

Arch. Biochem. Biophys.

309

77-80

1994

-

4

-

2

-

5

-

4

-

5

-

285245

Brodowsky

A linoleic acid (8R)-dioxygena ...

Gaeumannomyces graminis

J. Biol. Chem.

267

14738-14745

1992

-

1

-

1

-

1

-

1

-

1

-

Production of 7,8-dihydroxy unsaturated fatty acids from plant oils by whole recombinant cells expressing 7,8-linoleate diol synthase from Glomerella cingulata

Data extracted from this reference: