Molecular characterization of Penicillium oxalicum 6R,8R-linoleate diol synthase with new regiospecificity

Seo, Min‐Ju; Kang, Woo-Ri; Yang, Eun-Joo; Shin, Kyung‐Chul; Ko, Yoon‐Joo; Oh, Deok‐Kun

doi:10.1016/j.bbalip.2018.10.006

Cited by 5 publications

(15 citation statements)

References 43 publications

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“…The specific activity of recombinant E. coli expressing wild‐type 6,8‐LDS toward FFA followed the order, linoelic acid > oleic acid > palmitoleic acid > α‐linolenic acid (Table ), which is identical to the order of purified 6,8‐LDS reported in our previous study (Seo et al, ). However, recombinant E. coli expressing T879A variant 6,8‐LDS displayed the highest specific activity for oleic acid, among the FFA tested.…”

Section: Resultssupporting

confidence: 81%

“…In our previous study, we found that the T879A variant of 6,8‐LDS from P. oxalicum displayed 2.3‐fold higher activity than wild‐type 6,8‐LDS for linoelic acid (Seo et al, ). Thr879 residue in 6,8‐LDS from P. oxalicum was suggested to be located with substrate recognition site‐4 regions in the I‐helices of fungal LDS (Hoffmann and Oliw, ).…”

Section: Resultsmentioning

confidence: 91%

“…Maximum activity was observed at pH 6.5 and 35°C (Fig. S1, Supporting Information), while the maximum activity of the purified enzyme was observed at pH 6.5 and 25°C (Seo et al, ). Activities for the biotransformation of linoelic acid into DiHODE by recombinant E. coli expressing 5,8‐LDS from A. nidulans (Seo et al, ), 7,8‐LDS from G. cingulate (Seo et al, ), and 8,11‐LDS from P. chrysogenum (Kim et al, ) have been shown to be maximal at pH 7.5 and 35°C, pH 7.0 and 19°C, and pH 7.0 and 25°C, respectively.…”

Section: Resultsmentioning

confidence: 99%

“…Full‐length cDNA was synthesized from total RNA using a reverse transcription kit (TaKaRa, Shiga, Japan). The gene encoding 6,8‐LDS was cloned from cDNA, as previously described (Seo et al, ) by a one‐step isothermal DNA assembly method (Gibson et al, ). The genes encoding 6,8‐LDS and pET‐29b(+) expression vector were amplified using the following primers: forward primer for 6,8‐LDS gene, 5′‐AGC AGC GAA AAC CTG TAT TTT CAG GGA CAT ATG GTC GGA CAC CAC GGC AGC GAA ACC CAC‐3′; reverse primer for 6,8‐LDS gene, 5′‐ATC TCA GTG GTG GTG GTG GTG GTG CTC GAG TTA AGT GTA TCT CGC CTT GGG CAG CTC ACC‐3′; forward primer for expression vector, 5′‐GGT GAG CTG CCC AAG GCG AGA TAC ACT TAA CTC GAG CAC CAC CAC CAC CAC CAC TGA GAT‐3′; and reverse primer for expression vector, 5′‐GTG GGT TTC GCT GCC GTG GTG TCC GAC CAT ATG TCC CTG AAA ATA CAG GTT TTC GCT GCT‐3′.…”

Section: Methodsmentioning

confidence: 99%

“…Recently, the conversion of oleic acid, linoleic acid, and α‐linolenic acid into 6,8‐DiHOME, 6,8‐DiHODE, and 6,8‐DiHOTrE via 8‐HPOME, 8‐HPODE, and 8‐HPOTrE, respectively, by 6,8‐LDS from Penicillium oxalicum , has been reported (Fig. ) (Seo et al, ). However, the quantitative biotechnological production of 6,8‐DiHFA has not yet been performed.…”

Section: Introductionmentioning

confidence: 98%

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Production of 6,8‐Dihydroxy Fatty Acids by Recombinant Escherichia coli Expressing T879A Variant 6,8‐Linoleate Diol Synthase from Penicillium oxalicum

Shin

Seo

et al. 2019

J Americ Oil Chem Soc

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Recombinant Escherichia coli expressing T879A variant 6,8-linoleate diol synthase (LDS) from Penicillium oxalicum showed 2.1-fold higher activity than recombinant E. coli expressing wild-type 6,8-LDS for the production of 6,8-dihydroxy fatty acids (DiHFA) from linoleic acid. The optimal conditions for the production of 6,8-DiHFA by recombinant E. coli expressing T879A variant 6,8-LDS were pH 6.5, 35 C, 50 g L −1 cells, 10 g L −1 (35.7 mM) substrate, and 5% (v/v) dimethyl sulfoxide. Under these optimized conditions, 6.6 g L −1 (22.1 mM) 6,8-dihydroxy-9,12(Z,Z)octadecadienoic acid (DiHODE) and 7.1 g L −1 (22.6 mM) 6,8-dihydroxy-9(Z)-octadecenoic acid (DiHOME) were produced from linoleic acid and oleic acid in 40 min, respectively. The volumetric productivities of 6,8-DiHODE and 6,8-DiHOME under these conditions were 9.9 and 10.7 mg L −1 h −1 , respectively. The volumetric productivities of 6,8-DiHODE and 6,8-DiHOME were the highest values among those of all reported regiospecific DiHODE and DiHOME, respectively. To the best of our knowledge, this is the first quantitative biotechnological production of 6,8-DiHFA.Keywords 6,8-dihydroxy-9,12(Z,Z)-octadecadienoic acid Á 6,8-dihydroxy-9(Z)-octadecenoic Á 6,8-linoleate diol synthase Á Penicillium oxalicum J Am Oil Chem Soc (2019) 96: 663-669.

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Section: Resultssupporting

confidence: 81%

Section: Resultsmentioning

confidence: 91%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 98%

See 3 more Smart Citations

Production of 6,8‐Dihydroxy Fatty Acids by Recombinant Escherichia coli Expressing T879A Variant 6,8‐Linoleate Diol Synthase from Penicillium oxalicum

Shin

Seo

et al. 2019

J Americ Oil Chem Soc

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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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Biocatalytic oxyfunctionalization of unsaturated fatty acids to oxygenated chemicals via hydroxy fatty acids

Oh,

Lee,

Lee

et al. 2025

Biotechnology Advances

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Molecular characterization of Penicillium oxalicum 6R,8R-linoleate diol synthase with new regiospecificity

Cited by 5 publications

References 43 publications

Production of 6,8‐Dihydroxy Fatty Acids by Recombinant Escherichia coli Expressing T879A Variant 6,8‐Linoleate Diol Synthase from Penicillium oxalicum

Production of 6,8‐Dihydroxy Fatty Acids by Recombinant Escherichia coli Expressing T879A Variant 6,8‐Linoleate Diol Synthase from Penicillium oxalicum

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

Biocatalytic oxyfunctionalization of unsaturated fatty acids to oxygenated chemicals via hydroxy fatty acids

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