Mitsuo Miyazawa

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Organization: Kinki University , Japan

Department: Department of Applied Chemistry

Title: Professor(PhD)

TOPICS

Organic Chemistry

Chemicals
References

Chemical composition, aroma evaluation, and inhibitory activity towards acetylcholinesterase of essential oils from Gynura bicolor DC.

Co-reporter:Mitsuo Miyazawa;Hiroshi Nakahashi;Atsushi Usami

Journal of Natural Medicines 2016 Volume 70( Issue 2) pp:282-289

Publication Date(Web):2016 April

DOI:10.1007/s11418-015-0961-1

The compositions of the essential oils obtained from leaves and stems of Gynura bicolor DC. were analyzed by GC–MS. One hundred eight components of these oils were identified. (E)-β-caryophyllene (31.42 %), α-pinene (17.11 %), and bicyclogermacrene (8.09 %) were found to be the main components of the leaf oil, while α-pinene (61.42 %), β-pinene (14.39 %), and myrcene (5.10 %) were the major constituents of the stem oil. We found 73 previously unidentified components in these oils from G. bicolor. The oils were also subjected to odor evaluation. Eleven and 12 aroma-active compounds were detected in the leaf and stem oils, respectively. The abilities of these oils to inhibit acetylcholinesterase (AChE) activity were determined. The sesquiterpenoids in the oils were found to inhibit AChE activity more strongly than the monoterpenoids in the oils did. It was suggested that the three main components in each essential oil act synergistically against AChE activity. These results show that the essential oils obtained from G. bicolor are a good dietary source of AChE activity inhibition.

Chemical Composition and Character Impact Odorants in Volatile Oils from Edible Mushrooms

Co-reporter:Atsushi Usami;Ryota Motooka;Hiroshi Nakahashi;Shinsuke Marumoto

Chemistry & Biodiversity 2015 Volume 12( Issue 11) pp:1734-1745

Publication Date(Web):

DOI:10.1002/cbdv.201400395

Abstract

The aim of this study was to investigate the chemical composition and the odor-active components of volatile oils from three edible mushrooms, Pleurotus ostreatus, Pleurotus eryngii, and Pleurotus abalonus, which are well-known edible mushrooms. The volatile components in these oils were extracted by hydrodistillation and identified by GC/MS, GC-olfactometry (GC-O), and aroma extract dilution analysis (AEDA). The oils contained 40, 20, and 53 components, representing 83.4, 86.0, and 90.8% of the total oils in P. ostreatus, P. eryngii, and P. abalonus, respectively. Odor evaluation of the volatile oils from the three edible mushrooms was also carried out using GC-O, AEDA, and odor activity values, by which 13, eight, and ten aroma-active components were identified in P. ostreatus, P. eryngii, and P. abalonus, respectively. The most aroma-active compounds were C₈-aliphatic compounds (oct-1-en-3-ol, octan-3-one, and octanal) and/or C₉-aliphatic aldehydes (nonanal and (2E)-non-2-enal).

Aroma Evaluation of Setonojigiku (Chrysanthemum japonense var. debile) by Hydrodistillation and Solvent-assisted Flavour Evaporation

Co-reporter:Atsushi Usami;Hiroshi Nakahashi;Shinsuke Marumoto

Phytochemical Analysis 2014 Volume 25( Issue 6) pp:561-566

Publication Date(Web):

DOI:10.1002/pca.2528

ABSTRACT

Introduction

The Chrysanthemum genus consisting of about 200 species is mainly distributed over the Northern Hemisphere. Despite the pleasant odour of C. japonense var. debile (setonojigiku), no detailed analysis of the aroma-active compounds has been reported using sensory evaluation.

Objectives

Using a hydrodistillation (HD) and a solvent-assisted flavour evaporation (SAFE) method to obtain the volatile oil from the leaf parts.

Methods

To clarify odorants contributing to the characteristic aroma-active compounds, the aroma-extract dilution analysis (AEDA) method was performed through gas chromatography olfactometry (GC/O) analysis. In addition, the odour activity value (OAV) was calculated in order to determine the relative contribution of each compound to the aroma-active compounds.

Results

A total of 42 components by HD oil were identified by GC–MS, whereas 34 components were identified in SAFE oil. Thirteen compounds were identified by GC/O analysis in HD and SAFE oils respectively.

Conclusion

Each extraction method has its own advantages and disadvantages, and they are generally complementary to each other. On the basis of AEDA, OAV and sensory evaluations, [2.2.1] bicyclic monoterpenes (borneol, bornyl acetate and camphor) and β-caryophyllene are considered to be the main aroma-active compounds of both extraction methods. Copyright © 2014 John Wiley & Sons, Ltd.

Structure–activity relationships for naturally occurring coumarins as β-secretase inhibitor

Co-reporter:Shinsuke Marumoto, Mitsuo Miyazawa

Bioorganic & Medicinal Chemistry 2012 Volume 20(Issue 2) pp:784-788

Publication Date(Web):15 January 2012

DOI:10.1016/j.bmc.2011.12.002

The present study was demonstrated to evaluate the effects of naturally occurring coumarins (NOCs) including simple coumarins, furanocoumarins, and pyranocoumarins on the inhibition of β-secretase (BACE1) activity. Of 41 NOCs examined, some furanocoumarins inhibited BACE1 activity, but simple coumarins and pyranocoumarins did not affect. The most potent inhibitor was 5-geranyloxy-8-methoxypsoralen (31), which has an IC50 value of 9.9 μM. Other furanocoumarin derivatives, for example, 8-geranyloxy-5-methoxypsoralen (35), 8-geranyloxypsoralen (24), and bergamottin (18) inhibited BACE1 activity, with the IC50 values <25.0 μM. Analyses of the inhibition mechanism by Dixon plots and Cornish-Bowden plots showed that compounds 18, 31 and 35 were mixed-type inhibitor. The kinetics of inhibition of BACE1 by coumarins 24 was non-competitive inhibitors.

Aroma Evaluation of Gamazumi (Viburnum dilatatum) by Aroma Extract Dilution Analysis and Odour Activity Value

Co-reporter:Mitsuo Miyazawa;Shunsuke Hashidume;Toshiyuki Takahashi;Tohru Kikuchi

Phytochemical Analysis 2012 Volume 23( Issue 3) pp:208-213

Publication Date(Web):

DOI:10.1002/pca.1344

Introduction

Viburnum dilatatum (gamazumi) is widely distributed in Japan and China. Recently, juice from V. dilatatum fruits has been manufactured in Japan. Concerning the aroma of V. dilatatum, phenethyl alcohol, 3Z-hexenol and l-linalool have been identified in the essential oil from the flowers of V. dilatatum, however, there are no detailed reports on the aroma of V. dilatatum elucidated using sensory evaluation.

Objective

To clarify odourants contributing to the characteristic aroma, the aroma extract dilution analysis (AEDA) method was performed through gas chromatography olfactometry (GC-O) analysis.

Methodology

The aroma-active compounds were identified by GC-O and AEDA, and in order to determine the relative contribution of each compound to the aroma of V. dilatatum, odour activity value (OAV) has been used.

Results

The hydrodistillation of the leaf and branch of V. dilatatum afforded pale yellowish oils, with yields of 0.008 and 0.015% (w/w). The main components of the leaf oil were 3Z-hexenal (12.7%) and linalool (10.8%). In branch oil, palmitic acid (18.3%) and linoleic acid (8.2%) were identified. With regard to aroma components, 24 and 14 compounds were identified in the leaf and branch oils respectively, by GC-O analysis.

Conclusion

On the basis of AEDA, OAVs and sensory evaluations, nonanal is estimated as the main aroma compound of leaf and branch oil, as the other aroma compounds, C₆ compounds and 2-pentyl furan make green odour; linalool, eugenol and β-ionone play important role in the sweet odour of leaf oil. In branch oil, cis-furanlinalool oxide and eugenol make sweet odour, and β-eudesmol contributes to woody odour. Copyright © 2011 John Wiley & Sons, Ltd.

Corrigendum to: Two new aromatic compounds and a new d-arabinitol ester from the mushroom Hericium erinaceum [Tetrahedron 68 (2012) 2007–2010]

Co-reporter:Mitsuo Miyazawa, Toshiyuki Takahashi, Isao Horibe, Ryuuzou Ishikawa

Tetrahedron 2012 68(19) pp: 3786

Publication Date(Web):

DOI:10.1016/j.tet.2012.03.046

Two new aromatic compounds and a new d-arabinitol ester from the mushroom Hericium erinaceum

Co-reporter:Mitsuo Miyazawa, Toshiyuki Takahashi, Isao Horibe, Ryuuzou Ishikawa

Tetrahedron 2012 68(7) pp: 2007-2010

Publication Date(Web):

DOI:10.1016/j.tet.2011.11.068

Biotransformation of Turmerones by Aspergillus niger

Co-reporter:Mai Fujiwara, Shinsuke Marumoto, Nobuo Yagi, and Mitsuo Miyazawa

Journal of Natural Products 2011 Volume 74(Issue 1) pp:86-89

Publication Date(Web):December 28, 2010

DOI:10.1021/np100416v

Biotransformation studies conducted on (+)-(S)-ar-turmerone (1) and (+)-(S)-dihydro-ar-turmerone (2) by the fungus Aspergillus niger have revealed that 1 was metabolized to give four oxidized metabolites, (+)-(7S)-hydroxydehydro-ar-todomatuic acid (3), (+)-(7S,10E)-12-hydroxydehydro-ar-todomatuic acid (4), (+)-(7S,10E)-7,12-dihydroxydehydro-ar-todomatuic acid (5), and (+)-(7S)-15-carboxy-9,13-epoxy-7-hydroxy-9,13-dehydro-ar-curcumene (6), and (+)-(S)-dihydro-ar-turmerone (2) was metabolized to (+)-7,11-dihydroxy-ar-todomatuic acid (7). Metabolites 3−7 were characterized using spectroscopic techniques. Metabolites 3−7 inhibited acetylcholinesterase (AChE) although less so than the parent substrates.

Microbial reduction of coumarin, psoralen, and xanthyletin by Glomerella cingulata

Co-reporter:Shinsuke Marumoto, Mitsuo Miyazawa

Tetrahedron 2011 67(2) pp: 495-500

Publication Date(Web):

DOI:10.1016/j.tet.2010.10.089

Biotransformation of Bergapten and Xanthotoxin by Glomerella cingulata

Co-reporter:Shinsuke Marumoto and Mitsuo Miyazawa

Journal of Agricultural and Food Chemistry 2010 Volume 58(Issue 13) pp:7777-7781

Publication Date(Web):June 7, 2010

DOI:10.1021/jf101064v

The biotransformation of bergapten (1) by the fungus Glomerella cingulata gave the corresponding reduced acid, 6,7-furano-5-methoxy hydrocoumaric acid (2), a new compound. Xanthotoxin (3) was also converted to the corresponding reduced acid cnidiol b (4) and demethylated metabolite xanthotoxol (5) by G. cingulata. The structure of the new compound 2 was elucidated by high-resolution mass spectrometry, extensive NMR techniques, including 1H NMR and 13C NMR, 1H−1H correlation spectroscopy, heteronuclear multiple quantum coherence, and heteonuclear multiple bond coherence. The methyl ester or methyl ether or methyl ester and ether derivatives of 2 and 4 were synthesized. All compounds were tested for the β-secretase (BACE1) inhibitory activity in vitro. The methyl ester and ether derivative 8 was shown to possess BACE1 inhibitory activity, and a IC50 value was 0.64 ± 0.04 mM.

Biotransformation of isoimperatorin and imperatorin by Glomerella cingulata and β-secretase inhibitory activity

Co-reporter:Shinsuke Marumoto, Mitsuo Miyazawa

Bioorganic & Medicinal Chemistry 2010 Volume 18(Issue 1) pp:455-459

Publication Date(Web):1 January 2010

DOI:10.1016/j.bmc.2009.10.004

Biotransformation studies conducted on the furanocoumarins isoimperatorin (1) and imperatorin (3) have revealed that 1 was metabolized by Glomerella cingulata to give the corresponding reduced acid, 6,7-furano-5-prenyloxy hydrocoumaric acid (2), and 3 was transformed by G. cingulata to give the dealkylated metabolite, xanthotoxol (4) in high yields (83% and 81%), respectively. The structures of the new compound 2 have been established on the basis of spectral data. The metabolites 2 and 4 were tested for the β-secretase (BACE1) inhibitory activity in vitro, and metabolite 2 slightly inhibited the β-secretase activity with an IC50 value of 185.6 ± 6.8 μM. The metabolite 4 was less potent activity than compounds 1–3. In addition, methyl ester (2Me), methyl ether (2a) and methyl ester and ether (2aMe) of 2 were synthesized, and investigated for the ability to inhibit β-secretase. Compound 2aMe exhibited the best β-secretase inhibitory activity at the IC50 value 16.2 ± 1.2 μM and found to be the 2aMe showed competitive mode of inhibition against β-secretase with Ki value 11.3 ± 2.8 μM.

Suppression of SOS-Inducing Activity of Chemical Mutagens by Metabolites from Microbial Transformation of (+)-Longicyclene

Co-reporter:Kazuki Sakata and Mitsuo Miyazawa

Journal of Agricultural and Food Chemistry 2010 Volume 58(Issue 16) pp:9001-9005

Publication Date(Web):July 28, 2010

DOI:10.1021/jf101846p

In this study, biotransformation of (+)-longicyclene (1) by Aspergillus niger (NBRC 4414) and the suppressive effect on umuC gene expression by chemical mutagens 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide (furylfuramide) and aflatoxin B1 (AFB1) of the SOS response in Salmonella typhimurium TA1535/pSK1002 were investigated. Initially, compound 1 was converted to three new terpenoids, (−)-(10R)-10-hydroxy-longicyclic acid (2), (+)-(10S)-10-hydroxy-longicyclic acid (3), and (+)-10-oxo-longicyclic acid (4) by A. niger, and their conversion rates were 27, 23, and 30%, respectively. The metabolites suppressed the SOS-inducing activity of furylfuramide and AFB1 in the umu test. Compounds 1−4 were hardly showing a suppressive effect on umu gene expression of the SOS responses in S. typhimurium TA1535/pSK1002 against furylfuramid. However, metabolites showed a suppressive effect against AFB1. Compound 4 had gene expression by chemical mutagen AFB1, was suppressed 53% at <1.0 mM, and was the most effective compound in this experiment.

Regio- and Stereoselective Oxidation of (+)-Δ3-Carene by the Larvae of Common Cutworm (Spodoptera litura)

Co-reporter:Mitsuo Miyazawa and Haruki Kano

Journal of Agricultural and Food Chemistry 2010 Volume 58(Issue 6) pp:3855-3858

Publication Date(Web):February 26, 2010

DOI:10.1021/jf903301v

Biotransformation of (+)-Δ3-carene (1) and (+)-(1S,3S,4R,6R)-3,4-epoxycarane (1-1) by larvae of Spodoptera litura was investigated. Compound 1 was transformed to (+)-(1S,3S,4R,6R,7S)-3,4-epoxycaran-9-ol (1-2) by S. litura. This structure was established by infrared, electron impact−mass spectrometry, one- and two-dimensional NMR spectral studies, and (+)-(1S,3S,4R,6R)-3,4-epoxycarane (1-1) was transformed for confirmation of a metabolic pathway. The results indicate that the metabolic reaction of compound 1 by the larvae of S. litura was regioselective hydroxylation at the methyl group of the geminal dimethyl (C-9 position) followed by stereoselective epoxidation at the carbon−carbon double bands (C-3 position). (+)-(1S,3S,4R,6R,7S)-3,4-Epoxycaran-9-ol (1-2) was a new compound.

Acetylcholinesterase Inhibitory Activity of Volatile Oil from Peltophorum dasyrachis Kurz ex Bakar (Yellow Batai) and Bisabolane-Type Sesquiterpenoids

Co-reporter:Mai Fujiwara, Nobuo Yagi and Mitsuo Miyazawa

Journal of Agricultural and Food Chemistry 2010 Volume 58(Issue 5) pp:2824-2829

Publication Date(Web):February 10, 2010

DOI:10.1021/jf9042387

In this study, the chemical compositions and acetylcholinesterase (AChE) inhibitory activitiy of the volatile oil from the bark of Peltophorum dasyrachis Kurz ex Bakar (yellow batai) were evaluated. As a result, 68 compounds, accounting for 88.0% of the total oil, were identified. The main characteristic constituent in P. dasyrachis was isolated by silica gel column chromatography and found to be a sesquiterpenoid, (+)-(S)-ar-turmerone (1). In the AChE inhibitory assay, the volatile oil showed potent inhibitory activity with the IC50 value of 83.2 ± 2.8 μg/mL. Among the volatile oil components and characteristic sesquiterpenoids, (+)-(S)-ar-turmerone (1) and (+)-(S)-dihydro-ar-turmerone (2) were potent compounds, inhibiting AChE in a dose-dependent manner, with IC50 values of 191.1 ± 0.3 and 81.5 ± 0.2 μM, respectively. (+)-(S)-Dihydro-ar-turmerone (2), in particular, was found to be the most potent AChE inhibitor. Also, bisabolane-type sesquiterpenoid derivatives, (+)-(7S,9S)-ar-turmerol (3), (+)-(7S,9R)-ar-turmerol (4), (+)-(7S,9S)-dihydro-ar-turmerol (5), (+)-(7S,9R)-dihydro-ar-turmerol (6), (+)-(S)-ar-curcumene (7), and (+)-(S)-dihydro-ar-curcumene (8), were synthesized and tested for their AChE inhibitory effect, and their structure−activity relationships were evaluated. All sesquiterpenoids exhibited AChE inhibitory activity. The order of AChE inhibitory potency by bisabolane-type sesquiterpenoids was as follows: ketones > alcohols > hydrocarbons. Furthermore, the inhibition kinetics analyzed by Dixon plots indicated that (+)-(S)-ar-turmerone (1) is a competitive inhibitor, with a Ki value of 882.1 ± 2.1 μM, whereas (+)-(S)-dihydro-ar-turmerone (2) is a non-competitive inhibitor.

Suppression of SOS-Inducing Activity of Chemical Mutagens by Metabolites from Microbial Transformation of (−)-Isolongifolene

Co-reporter:Kazuki Sakata, Yoshimitsu Oda and Mitsuo Miyazawa

Journal of Agricultural and Food Chemistry 2010 Volume 58(Issue 4) pp:2164-2167

Publication Date(Web):January 28, 2010

DOI:10.1021/jf903651c

In this study, biotransformation of (−)-isolongifolene (1) by Glomerella cingulata and suppressive effect on umuC gene expression by chemical mutagens 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide (furylfuramide) and aflatoxin B1 (AFB1) of the SOS response in Salmonella typhimurium TA1535/pSK1002 were investigated. Initially, 1 was carried out the microbial transformation by G. cingulata. The result found that 1 was converted into (−)-isolongifolen-9-one (2), (−)-(2S)-13-hydroxy-isolongifolen-9-one (3), and (−)-(4R)-4-hydroxy-isolongifolen-9-one (4) by G. cingulata, and their conversion rates were 60, 25, and 15%, respectively. The metabolites suppressed the SOS-inducing activity of furylfuramid and AFB1 in the umu test. Comound 2 showed gene expression by chemical mutagens furylfuramide and AFB1 was suppressed 54 and 50% at <0.5 mM, respectively. Compound 2 is the most effective compound in this experiment.

Volatile compounds with characteristic odour in moso-bamboo stems (Phyllostachys pubescens Mazel ex Houz. De ehaie)

Co-reporter:Toshiyuki Takahashi;Kohji Mizui

Phytochemical Analysis 2010 Volume 21( Issue 5) pp:489-495

Publication Date(Web):

DOI:10.1002/pca.1224

Abstract

Introduction – Moso-bamboo (Phyllostachys pubescens) is well known as an edible shoot in Asia, and the stems of moso-bamboo are used as tableware due to its characteristic odour. Despite the pleasant odour of bamboo stems, no detailed analysis of the volatile compounds has been reported.

Objective – To clarify the potent odourants contributing to the characteristic aroma of the bamboo, the aroma extract dilution analysis (AEDA) method was performed through gas chromatography olfactometry (GC-O) analysis. In addition, relative flavour activity (RFA) was calculated, in which both the flavor dilution (FD) factor and weight percentage of each compound are involved.

Results – Eighty-nine compound in bamboo stems oil were identified by GC and GC-MS. The main components of the oil were palmitic acid (16.5%), (E)-nerolidol (10.2%) and indole (8.1%). In sensory analysis, 18 aroma-active compounds were detected by aroma extract dilution analysis (AEDA). The most intense aroma-active compounds were eugenol (sweet, clove-like, green) and (E)-2-nonenal (green).

Conclusion – The results of the sniffing test, RFA and FD factor indicated that (E)-2-nonenal and eugenol were estimated to have a bamboo-like aroma, and aldehyde compounds, such as a phenylacetaldehyde (floral) and C9–C10 unsaturated aldehydes, make the aroma of bamboo. Copyright © 2010 John Wiley & Sons, Ltd.

Regioselective Biotransformation of (+)- and (−)-Citronellene by the Larvae of Common Cutworm (Spodoptera litura)

Co-reporter:Mitsuo Miyazawa, Shinsuke Marumoto, Atsunori Masuda, Haruki Kano and Hiromune Takechi

Journal of Agricultural and Food Chemistry 2009 Volume 57(Issue 17) pp:7800-7804

Publication Date(Web):August 12, 2009

DOI:10.1021/jf9009069

Terpenoids, which have many biological activities and have occurred widely in nature, can be artificially synthesized. However, regioselective oxidation of terpenoids is difficult by chemical methods. In this study, (+)- and (−)-citronellene were biotransformed with Spodoptera litura to define the mechanism of metabolism of citronellene and gain a new natural terpenoid. (+)-Citronellene was converted to (2S,3S)-3,7-dimethyl-6-octene-1,2-diol and (2R,3S)-3,7-dimethyl-6-octene-1,2-diol (89.7%), (3S,6S)-(−)-3,7-dimethyl-1-octene-6,7-diol (3.8%), (3S)-(6E)-(+)-3,7-dimethyl-1,6-octadien-8-ol (4.2%), and (3S)-(6E)-(+)-3,7-dimethyl-1,6-octadien-8-oic acid (2.3%). In contrast, (−)-citronellene was converted to (2R,3R)-3,7-dimethyl-6-octene-1,2-diol and (2S,3R)-3,7-dimethyl-6-octene-1,2-diol (56.3%), (+)-iridan-7,8-diol (3.5%), and (3R)-(6E)-(−)-3,7-dimethyl-1,6-octadien-8-oic acid (40.2%). The main metabolic pathway of (+)- and (−)-citronellene by larvae of S. litura was oxidized at the terminal double bond and trans-allylic methyl position. Particularly on (+)-citronellene, the regioselective reaction was shown. On the oxidation of C-6, C-7, and C-8 positions, four new compounds (3S,6S)-(−)-3,7-dimethyl-1-octene-6,7-diol, (3S)-(6E)-(+)-3,7-dimethyl-1,6-octadien-8-oic acid, (+)-iridan-7,8-diol, and (3R)-(6E)-(−)-3,7-dimethyl-1,6-octadien-8-oic acid were produced in regioselective oxidation. It noted that stereoselective oxidation occurred between the enantiomers. The C-6 position was oxidized on the (+)-(3S) form, whereas cyclized and the C-7 position were oxidized on the (−)-(3R) form.

Biotransformation of three aromadendrane-type sesquiterpenoids by Aspergillus wentii

Co-reporter:Mitsuo Miyazawa;Tomohiko Takahashi;Kazuki Sakata ;Isao Horibe

Journal of Chemical Technology and Biotechnology 2008 Volume 83( Issue 7) pp:1006-1011

Publication Date(Web):

DOI:10.1002/jctb.1906

Abstract

BACKGROUND: The biotransformation of sesquiterpenoids, which are a large class of naturally occurring compounds, using microorganisms as a biocatalyst to produce useful novel organic compounds was investigated. The biotransformation of sesquiterpenoids, (+)-aromadendrene (1), (−)-alloaromadendrene (2) and (+)-ledene (3) has been investigated using Aspergillus wentii as a biocatalyst.

Results: Compound 1 was converted to (−)-(10S,11S)-10,13,14-trihydroxyaromadendrane (4). Compound 2 was converted to (+)-(1S,11S)-1,13-dihydroxyaromadendrene (5) and (−)-5,11-epoxycadin-1(10)-en-14-ol (6). Compound 3 was converted to compound 6, (+)-(10R,11S)-10,13-dihydroxyaromadendr-1-ene (7) and (+)-(10S,11S)-10,13-dihydroxyaromadendr-1-ene (8). The structure of the metabolic products has been elucidated on the basis of their spectral data.

CONCLUSION: Compound 1 gave only one product that was hydroxylated at C-10, C-13 and C-14. By contrast, compounds 2 and 3 gave a number of products, one of which was common. The differences in oxidation of 1–3 are due to the configuration of the C-1 position. Compounds 4–8 were new compounds. Copyright © 2008 Society of Chemical Industry

Biotransformation of isoflavones by Aspergillus niger as biocatalyst

Co-reporter:Mitsuo Miyazawa;Koji Takahashi;Hideo Araki

Journal of Chemical Technology and Biotechnology 2006 Volume 81(Issue 4) pp:674-678

Publication Date(Web):24 FEB 2006

DOI:10.1002/jctb.1461

Biotransformation of the isoflavones, 6,7,4′-trimethoxyisoflavone (1) and 5,7,4′-trimethoxyisoflavone (2) by Aspergillus niger was investigated. Compound 1 was transformed to 4′-hydroxy-6,7-dimethoxyisoflavone (3) and 2 to 4′-hydroxy-5,7-dimethoxyisoflavone (4). This suggested that 1 and 2 were demethylated at the C-4′ position with regioselectivity by Aspergillus niger. Copyright © 2006 Society of Chemical Industry

Microbial O-demethylation of sinesetin and antimutagenic activity of the metabolite

Co-reporter:Mitsuo Miyazawa;Yoshiharu Okuno

Journal of Chemical Technology and Biotechnology 2006 Volume 81(Issue 1) pp:29-33

Publication Date(Web):29 JUL 2005

DOI:10.1002/jctb.1353

Biotransformation of sinesetin by Aspergillus niger afforded 4′-hydroxy-5,6,7,3′-tetramethoxyflavone on the basis of its spectroscopic data including IR, heteronuclear multiple quantum coherence (HMQC) and heteronuclear multiple bond connectivity (HMBC) analysis. Sinesetin and the metabolite showed antimutagenic activity against chemical mutagens 4-dimethyl-3H-imidazo[4,5-f]quinolin-2-amine (MeIQ) and 3-amino-14-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) in the umu test using Salmonella typhimurium TA1535/pSK1002. Copyright © 2005 Society of Chemical Industry

Biotransformation of l-menthol by twelve isolates of soil-borne plant pathogenic fungi (Rhizoctonia solani) and classification of fungi

Co-reporter:Mitsuo Miyazawa;Hideki Kawazoe;Mitsuro Hyakumachi

Journal of Chemical Technology and Biotechnology 2003 Volume 78(Issue 6) pp:620-625

Publication Date(Web):14 APR 2003

DOI:10.1002/jctb.829

The microbial transformation of l-menthol (1) was investigated by using 12 isolates of soil-borne plant pathogenic fungi, Rhizoctonia solani (AG-1-IA Rs24, Joichi-2, RRG97-1; AG-1-IB TR22, R147, 110.4; AG-1-IC F-1, F-4, P-1; AG-1-ID RCP-1, RCP-3, and RCP-7) as a biocatalyst. Rhizoctonia solani F-1, F-4 and P-1 showed 89.7–99.9% yields of converted product from 1, RCP-1, RCP-3, and RCP-7 26.0–26.9% and the other isolates 0.1–12.0%. In the cases of F-1, F-4 and P-1, substrate 1 was converted to (−)-(1S,3R,4S,6S)-6-hydroxymenthol (2), (−)-(1S,3R,4S)-1-hydroxymenthol (3) and (+)-(1S,3R,4R,6S)-6,8-dihydroxymenthol (4), which was a new compound. Substrate 1 was converted to 2 and/or 3 by RRG97-1, 110.4, RCP-1, RCP-3 and RCP-7. The structures of the metabolic products were elucidated on the basis of their spectral data. In addition, metabolic pathways of the biotransformation of 1 by Rhizoctonia solani are discussed. Finally, from the main component analysis and the differences in the yields of converted product from 1, the 12 isolates of Rhizoctonia solani were divided into three groups based on an analysis of the metabolites. Copyright © 2003 Society of Chemical Industry

High-performance liquid chromatographic method for the determination of (−)-verbenone 10-hydroxylation catalyzed by rat liver microsomes

Co-reporter:Mitsuo Miyazawa, Atsushi Sugie, Tsutomu Shimada

Journal of Chromatography B 2003 Volume 793(Issue 2) pp:291-296

Publication Date(Web):15 August 2003

DOI:10.1016/S1570-0232(03)00329-5

A sensitive assay for the determination of (−)-verbenone 10-hydroxylation catalyzed by rat liver microsomes was developed using high-performance liquid chromatography. Verbenone was incubated in vitro with liver microsomes of untreated rats and rats treated with phenobarbital and the products thus formed were extracted with CH2Cl2 and the extracts were separated by HPLC with a C18 5-μm analytical column. Elution was conducted with 40% methanol containing 20 mM NaClO4 and the detection of UV absorbance was done at 251 nm. Product formation was dependent on the incubation time at least up to 30 min and the microsomal protein concentration between 0.01 and 0.1 mg protein/ml. The limit of detection of (−)-10-hydroxyverbenone with the HPLC was found to be about 40 pg, indicating that this method is about 100-fold sensitive than the GC–MS method. Optimized pH for the reaction was at 7.4 when examined with 100 mM potassium phosphate buffer in different pHs. Kinetic analysis showed that Km values for liver microsomes of untreated and phenobarbital-treated rats were 206 and 41 μM and Vmax values were 5.8 and 44 nmol/min/mg protein, respectively. Thus the present results provided a sensitive and useful method for the determination of verbenone 10-hydroxylation catalyzed by rat liver microsomes.

Biotransformation of l-menthol by soil-borne plant pathogenic fungi (Rhizoctonia solani)

Co-reporter:Hideki Kawazoe;Mitsuro Hyakumachi

Journal of Chemical Technology and Biotechnology 2002 Volume 77(Issue 1) pp:21-24

Publication Date(Web):28 NOV 2001

DOI:10.1002/jctb.522

The biotransformation of l-menthol was investigated by using nine isolates of Rhizoctonia solani (AG-1-IA Rs24, Joichi-2, RRG97-1; AG-1-IB TR22, R147, 110.4; AG-1-IC F-1, F-4 and P-1) as a biocatalyst. In the cases of Rhizoctonia solani F-1, F-4 and P-1, almost all of the substrate was consumed in 3 days and the major metabolite increased rapidly for the first of 3 days incubation. The structure of the major metabolite was elucidated on the basis of its spectral data. The major metabolite was determined to be (−)-(1S,3R,4S,6S)-6-hydroxymenthol which indicated that l-menthol was hydroxylated at the C-6 position. From the main component analysis, the nine isolates of Rhizoctonia solani were divided into two groups based on their ability to transform l-menthol to (−)-(1S,3R,4S,6S)-6-hydroxymenthol. This is the first report on the biotransformation of l-menthol by Rhizoctonia solani.

Biotransformation of (+)- and (−)-bornyl acetate using the plant parasitic fungus Glomerella cingulata as a biocatalyst

Co-reporter:Yoshiki Miyasato

Journal of Chemical Technology and Biotechnology 2001 Volume 76(Issue 2) pp:220-224

Publication Date(Web):23 JAN 2001

DOI:10.1002/jctb.367

The microbial transformations of (+)- and (−)-bornyl acetate were investigated using the plant parasitic fungus, Glomerella cingulata. As a result, (+)- and (−)-bornyl acetate were converted to (+)- and (−)-5-exo-hydroxybornyl acetate, (+)- and (−)-5-oxobornyl acetate and (+)- and (−)-borneol respectively. The structures of the metabolic products were determined by spectroscopic data.

Microbial resolution of racemic 2-endo-acetoxy-1,8-cineole by Glomerella cingulata

Co-reporter:Mitsuo Miyazawa, Yuya Hashimoto

Tetrahedron: Asymmetry 2001 Volume 12(Issue 22) pp:3185-3187

Publication Date(Web):10 December 2001

DOI:10.1016/S0957-4166(01)00555-9

Resolution of (±)-2-endo-acetoxy-1,8-cineole by Glomerella cingulata is described. Both (+)-2-endo-acetoxy-1,8-cineole and (−)-2-endo-hydroxy-1,8-cineole could be quantitatively obtained in enantiomerically pure form (yield 50%; e.e. 100%). In addition, the odor differences between the enantiomers are also described. In both compounds (acetoxy and hydroxy), the (+)-enantiomers tended to have more bright, light and sweet odors than their (−)-antipodes.Graphic(−)-(1R,2R,4S)-1,3,3-Trimethyl-2-oxabicyclo[2.2.2]octan-6-olC10H18O2E.e.=100% [determined by gas chromatography analysis on Cp-Cyclodextrin-β-236-M-19][α]D25 −19.6 (c 1.08 in CHCl3)Source of chirality: microbial resolutionAbsolute configuration: (1R,2R,4S)Acetate, (+)-(1S,2S,4R)-1,3,3-trimethyl-2-oxabicyclo[2.2.2]octan-6-olC12H20O3E.e.=100% [determined by gas chromatography analysis on Cp-Cyclodextrin-β-236-M-19][α]D25 +74.2 (c 0.96 in CHCl3)Source of chirality: microbial resolutionAbsolute configuration: (1S,2S,4R)

Biological stereoselective reduction of 4-methylcyclohexanone and 4-ethylcyclohexanone by anthracnose fungi

Co-reporter:Mitsuo Miyazawa;Shigeaki Okamura;Masashi Yamaguchi;Hiromu Kameoka

Journal of Chemical Technology and Biotechnology 2000 Volume 75(Issue 2) pp:143-146

Publication Date(Web):12 JAN 2000

DOI:10.1002/(SICI)1097-4660(200002)75:2<143::AID-JCTB195>3.0.CO;2-Z

The biotransformations of 4-methylcyclohexanone and 4-ethylcyclohexanone were investigated using 10 kinds of anthracnose fungi as biocatalysts. 4-Methylcyclohexanone and 4-ethylcyclohexanone were reduced to the corresponding cis- and trans-alcohols respectively. In the case of 4-methylcyclohexanone, it was transformed to mainly trans-4-methylcyclohexanol by all the fungi examined. In particular, the ratio of cis- and trans-alcohol products was shown to be 1:81 with high stereoselectivity by Colletotrichum lagenarium after a 7-day incubation period. The biotransformation of 4-ethylcyclohexanone by C lagenarium, C dematium MAFF410046, C trifolii MAFF305389, C fragariae, C atramentarium MAFF712102, C lindemuthianum (C-1), C lindemuthianum (C-3) and C lindemuthianum (C-13) produced mainly trans-4-ethylcyclohexanol. On the other hand, cis-alcohol was formed with stereoselectivity by Glomerella cingulata and C graminicola MAFF305460.

Biotransformation of (2Z,6Z)-farnesol by the plant pathogenic fungus Glomerella cingulata

Co-reporter:Hirokazu Nankai, Mitsuo Miyazawa, Hiromu Kameoka

Phytochemistry 1998 Volume 47(Issue 6) pp:1025-1028

Publication Date(Web):March 1998

DOI:10.1016/S0031-9422(98)80065-4

The microbial transformation of (2Z,6Z)-farnesol was investigated using the plant pathogenic fungus, Glomerella cingulata as a biocatalyst. Oxidation of the remote double bond and isomerization of the 2,3-double bond gave (2Z,6Z)-3,7,11-trimethyl-2,6-dodecadiene-1,10,11-triol and (2E,6Z)-3,7,11-trimethyl-2-6-dodecadiene-1,10,11-triol as major metabolites. One of the further degraded compounds, (Z)-9,10-dihydroxy-6,10-dimethyl-5-undecen-2-one, was also obtained.(2Z,6Z)-Farnesol (1) was transformed to two major metabolites, (2Z,6Z)-3,7,11-trimethyl-2,6-dodecadiene-1,10,11-triol (2) and (2E,6Z)-3,7,11-trimethyl-2,6-dodecadiene-1,10,11-triol (3) by Glomerella cingulata. (2Z,6Z)-3,7,11-trimethyl-2,6-dodecadiene-1,10,11-triol (Z)-9,10-Dihydroxy-6,10-dimethyl-5-undecen-2-one (4) was also obtained.

Resolution of racemic 2,6,6-trimethyl-7-oxa-bicyclo[3.1.1]octan-2-ol and 1,6,6-trimethyl-7-oxa-bicyclo[3.1.1]octan-2-ol by microbial esterification

Co-reporter:Mitsuo Miyazawa, Keisuke Tsuruno, Hiromu Kameoka

Tetrahedron: Asymmetry 1997 Volume 8(Issue 13) pp:2131-2132

Publication Date(Web):10 July 1997

DOI:10.1016/S0957-4166(97)00228-0

Resolution of racemic 2,6,6-trimethyl-7-oxa-bicyclo[3.1.1]octan-2-ol and racemic 1,6,6-trimethyl-7-oxa-bicyclo[3.1.1]octan-2-ol via esterification with malonic acid by Glomerella cingulata, is described. Both of alcohols and malonic esters could be obtained in enantiomerically pure states.The enantiomerically pure alcohols and malonic esters can be obtained by microbial esterification, Glomerella cingulata.

Biotransformation of (+)-(1R,2S,4R)-borneol and (−)-(1S,2R,4S)-borneol by Spodoptera litura (common cutworm) larvae

Co-reporter:Shinsuke Marumoto, Yoshiharu Okuno, Yohei Miyamoto, Mitsuo Miyazawa

Journal of Molecular Catalysis B: Enzymatic (May 2015) Volume 115() pp:160-167

Publication Date(Web):1 May 2015

DOI:10.1016/j.molcatb.2015.01.006

•The biotransformation of (+)- and (−)-borneols by insect Spodoptera litura larvae was investigated.•Eight new products were obtained.•A fatty acid conjugation reaction was the main metabolic pathway.In this study, we investigated the use of insects as biocatalysts for the biotransformation of monoterpenoids such as (+)-(1R,2S,4R)-borneol (1) and (−)-(1S,2R,4S)-borneol (2) by the larvae of Spodoptera litura. The substrates were administrated to the fourth instar larvae of S. litura through food, and the metabolites were collected in the feces. Analysis showed that substrate 1 was converted into the following 12 metabolites: (+)-(1R,2S,4R)-borneol linoleate (1–1), (+)-(1R,2S,4R)-borneol linolenate (1–2), (+)-(1R,2S,4R)-borneol-2-O-β-d-glucopyranoside (1–3), (+)-(1R,2S,4R,5S)-5-endo-hydroxyborneol (1–4), (+)-(1R,4R,5S)-5-endo-hydroxycamphor (1–5), (+)-(1R,2S,4R,5R)-5-exo-hydroxyborneol (1–6), (+)-(1R,4R,5R)-5-exo-hydroxycamphor (1–7), (+)-(1R,2S,4R,7R)-8-hydroxyborneol (1–8), (+)-(1R,4R,7R)-8-hydroxycamphor (1–9), (+)-(1R,2S,4R,7S)-9-hydroxyborneol (1–10), (+)-(1R,2S,4R)-10-hydroxyborneol (1–11), and (+)-(1R,2R,3S,4S)-3-endo-hydroxyborneol (1–12). The metabolism of substrate 2 generated 12 enantiomers of the above metabolites, namely 2–1 to 2–12. Compounds 1–1, 1–2, 1–3, 2–1, 2–2, 2–8, 2–10, and 2–12 are novel compounds. The main metabolites were generated by phase II reactions such as esterification or glucosidation of the parent substrates. The minor metabolites originated from phase I reactions such as hydroxylation or oxidation, probably by cytochrome P450 enzymes. These fatty acid and glucose conjugation reactions are new metabolic pathways, compared with those reported in previous studies on monoterpenoids in S. litura.Download full-size image

Stereoselective biocatalytic reduction of α-ionone by Glomerella cingulata

Co-reporter:Mitsuo Miyazawa, Kazuya Shimizu

Journal of Molecular Catalysis B: Enzymatic (January 2012) Volume 74(Issues 1–2) pp:6-8

Publication Date(Web):1 January 2012

DOI:10.1016/j.molcatb.2011.07.021

The biotransformation of (±)-α-ionone (1) by Glomerella cingulata was investigated. Compound 1 was transformed into two compounds (2,3). The ketone was reduced to α-ionol and the olefin was reduced to dihydrio-α-ionol, respectively. (−)-(6S,9R)- and (+)-(6R,9S)-α-ionol proceeded the corresponding allylic alcohols in high enantiomeric excess >99%; (−)-(S)-α-ionone is preferentially metabolized by hydroxylation in the ketone at C-9. Especially on the metabolic pathway, olefine reduction was via after ketone reduction.Graphical abstractDownload full-size imageHighlights► The biotransformation of (±)-α-ionone (1) by Glomerella cingulata was investigated. ► Biocatalytic reactions are often highly enanioselective at the corresponding allylic alcohols. ► Especially, on the metabolic pathway, this conversion route was unique.