Microbial conversion of 4-oxoisophorone by Aspergillus niger.

Yamazaki, Yoshinari; Hayashi, Yumiko; Hori, Nobuaki; Mikami, Yoichi

doi:10.1271/bbb1961.52.2919

Cited by 11 publications

(5 citation statements)

References 3 publications

(3 reference statements)

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“…6) However, there has been no report on the enzyme involved in the KIP reduction in these strains. In this study, we screened for microorganisms catalyzing the reduction of KIP to (6R)-levodione.…”

Section: Discussionmentioning

confidence: 99%

“…Microbial production of (6R)-levodione through stereospeciˆc C＝C bond reduction of a prochiral compound, 3,5,5-trimethyl-2-cyclohexene-1,4-dione (ketoisophorone; KIP), has been reported by several researchers. 2,[5][6][7] However, the enzyme catalyzing the reduction of the C＝C bond of KIP in these microorganisms has not been isolated or characterized. Two enone reductases catalyzing the asymmetric reduction of KIP to (6R)-levodione have been isolated from Saccharomyces cerevisiae, 8) although no structural analysis of the enzymes was reported.…”

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confidence: 99%

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Old Yellow Enzyme fromCandida macedoniensisCatalyzes the Stereospecific Reduction of the C=C Bond of Ketoisophorone

Kataoka

Kotaka

Hasegawa

et al. 2002

Bioscience, Biotechnology, and Biochemistry

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

Old Yellow Enzyme fromCandida macedoniensisCatalyzes the Stereospecific Reduction of the C=C Bond of Ketoisophorone

Kataoka

Kotaka

Hasegawa

et al. 2002

Bioscience, Biotechnology, and Biochemistry

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“…Two-step conversion of 2,6,6-trimethyl-2-cyclohexen-1,4-dione (ketoisophorone) to 4-hydroxy-2,2,6-trimethylcyclohexanone ( Fig. 1) by bacterial cells has been reported previously (3,11,12,19). However, in those cases, mixtures of isomers, (4R,6S), (4S,6R), (4R,6R), and (4S,6S), were produced and the productivity was low (ϳ2.5 mg/ml).…”

mentioning

confidence: 78%

“…Microbial production of actinol from ketoisophorone has been reported previously (3,11,12,19); however, in those cases, a racemic mixture of 4-hydroxy-2,2,6-trimethylcyclohexanone was obtained as the reduction product, and the enzymes involved in reduction of ketoisophorone and levodione have not yet been purified. In this study, we have established an overexpression system for S. cerevisiae OYE in E. coli, and the system provides a powerful tool for obtaining optically pure actinol, together with the LVR overexpression system established previously (21).…”

Section: Discussionmentioning

confidence: 99%

Production of a Doubly Chiral Compound, (4 R ,6 R )-4-Hydroxy-2,2,6-Trimethylcyclohexanone, by Two-Step Enzymatic Asymmetric Reduction

Wada

Yoshizumi

Noda

et al. 2003

Appl Environ Microbiol

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A practical enzymatic synthesis of a doubly chiral key compound, (4R,6R)-4-hydroxy-2,2,6-trimethylcyclohexanone, starting from the readily available 2,6,6-trimethyl-2-cyclohexen-1,4-dione is described. Chirality is first introduced at the C-6 position by a stereoselective enzymatic hydrogenation of the double bond using old yellow enzyme 2 of Saccharomyces cerevisiae, expressed in Escherichia coli, as a biocatalyst. Thereafter, the carbonyl group at the C-4 position is reduced selectively and stereospecifically by levodione reductase of Corynebacterium aquaticum M-13, expressed in E. coli, to the corresponding alcohol. Commercially available glucose dehydrogenase was also used for cofactor regeneration in both steps. Using this two-step enzymatic asymmetric reduction system, 9.5 mg of (4R,6R)-4-hydroxy-2,2,6-trimethylcyclohexanone/ml was produced almost stoichiometrically, with 94% enantiomeric excess in the presence of glucose, NAD ؉ , and glucose dehydrogenase. To our knowledge, this is the first report of the application of S. cerevisiae old yellow enzyme for the production of a useful compound.(4R,6R)-4-hydroxy-2,2,6-trimethylcyclohexanone (actinol) is an ideal precursor for the synthesis of naturally occurring, optically active hydroxylated carotenoids, such as zeaxanthin (8), cryptoxanthin, and structurally related compounds (1, 2). Two-step conversion of 2,6,6-trimethyl-2-cyclohexen-1,4-dione (ketoisophorone) to 4-hydroxy-2,2,6-trimethylcyclohexanone ( Fig. 1) by bacterial cells has been reported previously (3,11,12,19). However, in those cases, mixtures of isomers, (4R,6S), (4S,6R), (4R,6R), and (4S,6S), were produced and the productivity was low (ϳ2.5 mg/ml). The enzymes involved in these reactions have not yet been identified.The discovery, purification, and characterization of levodione reductase (LVR), which catalyzes the regio-and stereospecific reduction of (6R)-2,2,6-trimethylcyclohexane-1,4-dione [(6R)-levodione] to actinol, has been reported (17). This LVR has also been cloned and expressed in Escherichia coli (21). Recently, it was also reported that old yellow enzyme (OYE; EC 1.6.99.1) of Candida macedoniensis catalyzes the stereospecific hydrogenation of ketoisophorone to (6R)-levodione (4). However, this enzyme has not yet been cloned, and enzyme purification from C. macedoniensis is needed for (6R)-levodione production. Thus, we tried to use OYE of Saccharomyces cerevisiae, for which the whole-genome sequence is now available. S. cerevisiae has two OYEs: OYE2, encoded by the OYE2 gene (15), and OYE3, encoded by the OYE3 gene (10). Both of them were cloned and expressed in E. coli, and the conversion efficiencies of ketoisophorone to (6R)-levodione were then compared.Here, we report actinol production by the stereoselective hydrogenation of the double bond of ketoisophorone and by the stereoselective reduction of the carbonyl group at the C-4 position of (6R)-levodione. For the first step, hydrogenation, the OYE2 of S. cerevisiae expressed in E. coli was used, and for the second step, carbon...

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“…Both substrates and products of the ER reaction can be subject to DHs-catalysed reduction thereby reducing yields and complicating product isolation. In particular using whole cells, this may be an issue as exemplified in the reduction of oxoisophorone by Aspergillus niger 369 (Fig. 22) and other organisms.…”

Section: Reduction Of C C Double Bondsmentioning

confidence: 99%