Identification of Dioxygenases Required for Aspergillus Development

Garscha, Ulrike; Jernerén, Fredrik; Chung, Dawoon; Keller, Nancy P.; Hámberg, Mats; Oliw, Ernst H.

doi:10.1074/jbc.m705366200

Cited by 90 publications

(74 citation statements)

References 41 publications

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“…formed by antarafacial dioxygenation at C-8 and suprafacial hydroxylation at C-7 and C-11. This mechanism of biosynthesis appears to be a characteristic feature of the family of LDS enzymes (7,16,42). We conclude that the metabolites discussed above can be formed by 7,8-LDS, as summarized in Fig.…”

Section: Discussionmentioning

confidence: 82%

“…(8R)-HODE and (5S,8R)-DiHODE were found to induce premature sexual sporulation (20). Gene deletion of 5,8-LDS and (10R)-DOX in A. nidulans and A. fumigatus was later found to influence sporulation, development, mycotoxin production, and pathogenicity (16,21,22). In contrast, little is known about the biological function of 7,8-LDS of G. graminis.…”

Section: (8r)-hpode Is Isomerized To (5s8r)-dihode By 58-lds and mentioning

confidence: 99%

“…)-DiHODE appear to be formed from (8R)-HPODE by the same mechanism as (7S,8S)-DiHODE (7,16,19), viz. suprafacial hydrogen abstraction and hydroxylation.…”

Section: (8r)-hpode Is Isomerized To (5s8r)-dihode By 58-lds and mentioning

confidence: 99%

“…Prostaglandin F 1␣ (100 ng/min) was infused for tuning. Products formed from stereospecifically deuterated 18:2n-6 were analyzed as described previously (16), and the deuterium content of the recovered 18:2n-6 was determined (LC-MS/MS analysis; m/z 276 -2823 full scan). For preparative use, we employed a large column (20 ϫ 150-mm Reprosil 100 ODS-A, 5 ; Dr. Maisch, Ammerbuch, Germany), eluted with 85-90% methanol in water with 0.1% acetic acid.…”

Section: Pcr Analysis Of Gene Transcripts In M Oryza the ⌬Mac1 Mutamentioning

confidence: 99%

“…or (10R)-HPODE by abstraction of the pro-S hydrogen at C-8 followed by antarafacial oxygen insertion at C-8 by LDS or at C-10 with double bond migration in the case of (10R)-DOX (16)…”

mentioning

confidence: 99%

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Gene Deletion of 7,8-Linoleate Diol Synthase of the Rice Blast Fungus

Jernerén

Sesma

Francheschetti

et al. 2010

Journal of Biological Chemistry

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Section: Discussionmentioning

confidence: 82%

Section: (8r)-hpode Is Isomerized To (5s8r)-dihode By 58-lds and mentioning

confidence: 99%

“…)-DiHODE appear to be formed from (8R)-HPODE by the same mechanism as (7S,8S)-DiHODE (7,16,19), viz. suprafacial hydrogen abstraction and hydroxylation.…”

Section: (8r)-hpode Is Isomerized To (5s8r)-dihode By 58-lds and mentioning

confidence: 99%

Section: Pcr Analysis Of Gene Transcripts In M Oryza the ⌬Mac1 Mutamentioning

confidence: 99%

“…or (10R)-HPODE by abstraction of the pro-S hydrogen at C-8 followed by antarafacial oxygen insertion at C-8 by LDS or at C-10 with double bond migration in the case of (10R)-DOX (16)…”

mentioning

confidence: 99%

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Gene Deletion of 7,8-Linoleate Diol Synthase of the Rice Blast Fungus

Jernerén

Sesma

Francheschetti

et al. 2010

Journal of Biological Chemistry

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Production of 8,11‐dihydroxy fatty acids from oleic and palmitoleic acids by Escherichia coli cells expressing variant 6,8‐linoleate diol synthases from Penicillium oxalicum

Shin

Seo

Kang

et al. 2022

Biotechnology Progress

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Recombinant Escherichia coli cells expressing 8,11‐linoleate diol synthase (LDS) from Penicillium chrysogenum convert oleic and palmitoleic acids to 8‐hydroperoxy‐9(Z)‐octadecenoic acid (HPOME) and 8‐hydroperoxy‐9(Z)‐hexadecenoic acid (HPHME) only, respectively. However, recombinant E. coli cells expressing Q889A variant 6,8‐LDS from Penicillium oxalicum as an 8,11‐LDS converted oleic and palmitoleic acids to 8,11‐dihydroxy‐9(Z)‐octadecenoic acid (DiHOME) and 8,11‐dihydroxy‐9(Z)‐hexadecenoic acid (DiHHME), respectively, which were identified using liquid chromatography–tandem mass spectrometry analysis. To select suitable variants for producing these compounds, position 889 of 6,8‐LDS from P. oxalicum was substituted with other amino acids, and recombinant E. coli cells expressing Q889L and Q889A variants were chosen as the best biocatalysts for producing 8,11‐DiHOME and 8,11‐DiHHME, respectively. The optimal conditions for producing 8,11‐DiHOME or 8,11‐DiHHME using cells expressing Q889L or Q889A variant 6,8‐LDS were pH 6.5 and 30 °C with 5% (v/v) dimethyl sulfoxide, 60 g L−1 cells, and 10 g L−1 oleic acid or 7.5 g L−1 palmitoleic acid, respectively. Under these conditions, 10.7 g L−1 8,11‐DiHOME and 8.1 g L−1 8,11‐DiHHME were produced for 1.5 h with molar yields of 96.4% and 96.2% and productivities of 7.1 and 5.4 g L−1 h−1, respectively. The molar yields and concentrations of 8,11‐DiHOME and 8,11‐DiHHME were highest among those of other reported DiHOMEs and DiHHMEs. To the best of our knowledge, this is the first quantitative production of 8,11‐DiHOME and 8,11‐DiHHME.

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Influence of Substrate Dideuteration on the Reaction of the Bifunctional Heme Enzyme Psi Factor Producing Oxygenase A (PpoA)

et al. 2011

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PpoA is a bifunctional enzyme that catalyzes the dioxygenation of unsaturated C18 fatty acids. The products of this reaction are termed psi factors and have been shown to play a crucial role in conferring a balance between sexual and asexual spore development as well as production of secondary metabolites in the fungus Aspergillus nidulans. Studies on the reaction mechanism revealed that PpoA uses two different heme domains to catalyze two subsequent reactions. Initially, the fatty acid substrate is dioxygenated at C8, yielding an 8-hydroperoxy fatty acid at the N-terminal domain. This reaction is catalyzed by a peroxidase/dioxygenase-type domain that exhibits many similarities to prostaglandin H2 synthases and involves a stereospecific homolytic hydrogen abstraction from C8 of the substrate. The C terminus harbors a heme thiolate P450 domain in which rearrangement of the 8-hydroperoxide to the final product, a 5,8-dihydroxy fatty acid, takes place. To obtain further information about the intrinsic kinetics and reaction mechanism of PpoA, we synthesized C5-dideutero- and C8-dideutero-oleic acid by a novel protocol that offers a straightforward synthesis without employing the toxic additive hexamethylphosphoramide (HMPA) during CC coupling reactions or mercury salts upon thioketal deprotection. These deuterated fatty acids were then employed for kinetic analysis under multiple-turnover conditions. The results indicate that the hydrogen abstraction at C8 is the rate-determining step of the overall reaction because we observed a KIE (V(H) /V(D) ) of ∼33 at substrate saturation that suggests extensive nuclear tunneling contributions for hydrogen transfer. Deuteration of the substrate at C5, however, had little effect on V(H) /V(D) but resulted in a different product pattern presumably due to an altered lifetime and partitioning of a reaction intermediate.

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Identification of Dioxygenases Required for Aspergillus Development

Cited by 90 publications

References 41 publications

Gene Deletion of 7,8-Linoleate Diol Synthase of the Rice Blast Fungus

Gene Deletion of 7,8-Linoleate Diol Synthase of the Rice Blast Fungus

Production of 8,11‐dihydroxy fatty acids from oleic and palmitoleic acids by Escherichia coli cells expressing variant 6,8‐linoleate diol synthases from Penicillium oxalicum

Influence of Substrate Dideuteration on the Reaction of the Bifunctional Heme Enzyme Psi Factor Producing Oxygenase A (PpoA)

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