Stereochemistry of matairesinol formation by <Emphasis Type="Italic">Daphne</Emphasis> secoisolariciresinol dehydrogenase

Secoisolariciresinol dehydrogenase activity was detected for the first time from Daphne odora and Daphne genkwa (Thymelaeaceae), which are known to produce optically pure (+)-matairesinol. In sharp contrast, (–)-matairesinol was formed selectively over the (+)-antipode by the secoisolariciresinol dehydrogenase preparation from both D. odora callus and D. genkwa shoots.     

Enantiomeric compositions and biosynthesis ofWikstroemia sikokiana lignans

Thymelaeaceae plants produce dextrorotatory dibenzylbutyrolactone lignans, which are opposite enantiomers to the lignans isolated from other plants (e.g.,Forsythia spp.). In our previous paper, (–)-pinoresinol (74% enantiomer excess), (+)-matairesinol (optically pure), and (+)-wikstromol (optically pure) were isolated fromWikstroemia sikokiana (Thymelaeaceae). In the present investigation, a survey of lignans and the determination of their enantiomeric compositions were continued. Four lignans, (–)-lariciresinol, (–)-secoisolariciresinol, (+)-kusunokinin, and (+)-methyltrachelogenin, were isolated from MeOH extracts ofW. sikokiana stem. To our knowledge, we have isolated (+)-methyltrachelogenin from plants for the first time. Chiral high-performance liquid chromatographic analysis showed that (+)-kusunokinin and (+)-methyltrachelogenin were optically pure, whereas (–)-lariciresinol and (–)-secoisolariciresinol were not (39% and 45% enantiomer excess, respectively). Feeding experiments with deuterium-labeled substrates demonstrated conversion of coniferyl alcohol to the lignans and interconversion of lignans. These reaction sequences are similar to the sequence catalyzed byForsythia enzymes. However, predominant enantiomers of the lignans, except for secoisolariciresinol isolated fromW. sikokiana, have absolute configurations opposite to those of the corresponding lignans isolated fromForsythia spp. Based on the results of the isolation and the feeding experiments, several differences betweenW. sikokiana andForsythia spp. are pointed out regarding stereochemical mechanisms for lignan biosynthesis.Parts of this report were presented at the 46th annual meeting of the Japan Wood Research Society, Kumamoto, April 1996; and the 47th annual meeting of the Japan Wood Research Society, Kochi, April 1997     

Lignan production inDaphne odora cell cultures

Lignan production in callus and cell suspension cultures ofDaphne odora is reported for the first time. The cell suspension culture produced pinoresinol, lariciresinol, secoisolariciresinol, matairesinol, and wikstromol. The production of matairesinol in the cell suspension culture was much higher than that inDaphne odora stem tissues.Part of this report was presented at the 51th annual meeting of the Japan Wood Research Society, Tokyo, April 2001     

Lignans ofLinum flavum var.compactum

A new dibenzylbutyrolactone lignan 7.6-dihydroxybursehernin, together with six known lignans (pinoresinol, lariciresinol, secoisolariciresinol,-peltatin,-peltatin, 5-methoxypodophyllotoxin) were isolated from the methanol extracts ofLinum flavum var.compactum. The enantiomeric analysis of pinoresinol and lariciresinol isolated from the species, which are upstream lignans in the lignan biosynthetic pathway, indicated that they are not optically pure, which is in accordance with our recent findings on lignans occurring in other plant species.Parts of this report were presented at the 42nd Lignin Symposium, Sapporo, October 1997; and the 43rd Lignin Symposium, Fuchu, October 1998     

Lignans ofChamaecyparis obtusa

Heartwood ofChamaecyparis obtusa contains significant amounts of a dibenzylbutyrolactone lignan, hinokinin (8). This investigation demonstrated that the contents of 8 and a norlignan, hinokiresinol (12), were higher in the heartwood region than in the sapwood, indicating their nature of being heartwood extractives. Eleven lignans — xanthoxylol (1), 7-oxohinokinin (2), savinin (3), dihydrosesamin (4), isoactifolin (5), sesamin (6), piperitol (7), hinokinin (8), pluviatolide (9), haplomyrfolin (10), and rnatairesinol (11) — were isolated from young shoots ofChamaecyparis obtusa cv. Breviramea. Eight lignans (1, 2, 4, 5, 7, 9,10, and11) were isolated from this plant for the first time. Chiral high-performance liquid Chromatographie analysis showed that8, 9, 10, and11, were found to be levorotatory and optically pure (>99% e.e.). Based on the chemical structures of the isolated lignans, possible biosynthetic pathways of8 are discussed.Parts of this report were presented at the 44th annual meeting of the Japan Wood Research Society, Nara, April 1994; the 46th annual meeting of the Japan Wood Research Society, Kumamoto, April 1996; and the 44th Lignin Symposium, Gifu, October 1999     

Survey and enzymatic formation of lignans ofAnthriscus sylvestris

Gas chromatography — mass spectrometry analysis of the -glucosidase-treated MeOH extracts ofAnthriscus sylvestris showed, based on comparison of the mass spectra and retention times with those of authentic samples, the presence of lignans, yatein, secoisolariciresinol, lariciresinol, matairesinol, hinokinin, and pluviatolide. The existence of small amounts of bursehernin was suggested by mass chromatography. In addition, nemerosin and deoxypodophyllotoxin were tentatively identified by comparing the mass spectra with those reported in the literature. Enzyme preparations fromA. sylvestris catalyzed the formation of secoisolariciresinol and lariciresinol from coniferyl alcohol. Furthermore, the enzyme preparation catalyzed the formation of lariciresinol from (±)-pinoresinols and the formation of secoisolariciresinol from (±)-lariciresinols. Thus, pinoresinol/lariciresinol reductase (PLR) activity was detected. Chiral high-performance liquid chromatography analysis showed selective formation of (+)-lariciresinol and (–)-secoisolariciresinol from (±)pinoresinols with theA. sylvestris PLR preparation, indicating that the stereochemical property ofA. sylvestris PLR-catalyzed reduction was similar to those ofForsythia PLR andArctium lappa ripening seed PLR.Part of this report was presented at the 43rd Lignin Symposium, Fuchu, October 1998     

Phenolic constituents ofTaxus cuspidata I: lignans from the roots

The phenolic constituents of the roots ofTaxus cuspidata (Japanese yew) were investigated. Four lignans, [(+)-taxiresinol (1), (+)-lariciresinol (2), (–)-secoisolariciresinol (3), and (+)-pinoresinol (4)] were isolated and identified. The assignment of proton and carbon atoms for the lignans were finally solved by one- and twodimensional-nuclear magnetic resonance spectra. The enantiomeric excess of these lignans were determined by chiral high-performance liquid Chromatographic analyses. (+)-Lariciresinol and (–)-secoisolariciresinol were optically pure; (+)-taxiresinol was also suggested to be optically pure, although (+)-pinoresinol was not (77% enantiomeric excess).     

Stereochemistry and biosynthesis of (+)-lyoniresinol, a syringyl tetrahydronaphthalene lignan in Lyonia ovalifolia var. elliptica I: isolation and stereochemistry of syringyl lignans and predicted precursors to (+)-lyoniresinol from wood

Steps leading to the biosynthesis of syringyl lignans and tetrahydronaphthalene and naphthalene lignans, especially the formation of the C2–C7′ linkage, have not been elucidated. Lyoniresinol is a typical syringyl lignan, as well as a tetrahydronaphthalene lignan found in Lyonia ovalifolia var. elliptica. To demonstrate the biosynthetic pathway for (+)-lyoniresinol, three putative biosynthetic intermediates of lyoniresinol, syringaresinol, 5,5′-dimethoxylariciresinol, and 5,5′-dimethoxysecoisolariciresinol, were isolated from wood. The identity of the putative intermediates was confirmed by spectroscopic analyses, as well as by comparison of spectral and chromatographic data with those of authentic samples previously synthesized. The stereochemistry (enantiomeric composition and absolute configuration) of the isolated lignans were determined as (±)-syringaresinol, (8S,8′S)-(−)-5,5′-dimethoxylariciresinol [46% enantiomeric excess (e.e.)], (8S,8′S)-(+)-5,5′-dimethoxysecoisolariciresinol (91% e.e.), and (8R,8′R)-(+)-lyoniresinol (42% e.e.). The absolute configurations of (+)-and (-)-5,5′-dimethoxylariciresinols, and (+)-and (-)-5,5′-dimethoxysecoisolariciresinols were determined by their synthesis (catalytic reduction) from (8R,8′R)-(+)-and (8S,8′S)-(-)-syringaresinols and by subsequent chiral high-performance liquid chromatography analysis. This report was presented at the 55th Annual Meeting of the Japan Wood Research Society, Kyoto, March 2005     

Biosynthesis of (+)-syringaresinol inLiriodendron tulipifera I: feeding experiments withl-[U-14C]phenylalanine and [8-14C]sinapyl alcohol

To clarify the biosynthesis of syringyl lignans and lignan formation by stereoselective coupling of monolignols, formation of (+)-syringaresinol and (+)-pinoresinol inLiriodendron tulipifera were investigated by means of feeding experiments. Following individual administration ofl-[U-¹⁴C]phenylalanine and [8-¹⁴C]sinapyl alcohol to excised shoots ofL. tulipifera and their subsequent metabolism for 3h, free [¹⁴C] lignans and [¹⁴C] lignan glucosides were extracted from both of the stems and leaves with methanol and divided into an ether fraction and an aqueous one, respectively. The glucosides were hydrolyzed by a combination of cellulase and-glucosidase to liberate [¹⁴C]lignans as aglycones.l-[U-¹⁴C]Phenylalanine was incorporated into free (+)-[¹⁴C]syringaresinol and its glucosides; the (+)-[¹⁴C]syringaresinols in the stems and leaves had 52% enantiomeric excess (% e.e.) and 42% e.e., respectively; and the (+)-[¹⁴C]syringaresinol aglycones from the glucosides in the stems and leaves had 20% e.e. and 22% e.e., respectively. Furthermore, [8-¹⁴C]sinapyl alcohol was incorporated into (+)-[¹⁴C]syringaresinol and its glucosides in the stems. These results suggest that the (+)-enantiomer of syringaresinol was enantioselectively formed from two molecules of sinapyl alcohol inL. tulipifera followed by transformation into the (+)-syringaresinol glucosides, accompanying the formation of racemic syringaresinol by nonselective coupling and the subsequent transformation of the racemate into their glucosides.l-[U-¹⁴C]Phenylalanine was incorporated also into free (+)-[¹⁴C]pinoresinol and its glucosides with 12%–42% e.e.Part of this paper was presented at the 47th Annual Meeting of the Japan Wood Research Society, Kochi, April 1997     

Abietane-type and labdane-type diterpenoids from the cones of Chamaecyparis obtusa

A novel compound, 8(17),12E,14-labdatrien-19-al (trans-communal), was isolated from ethyl acetate extract of young cones of hinoki (Chamaecyparis obtusa Endl.). The chemical structure of the compound was determined mainly with various nuclear magnetic resonance spectral techniques. Its stereochemistry was determined by derivation to a known compound,trans-communol. In addition to this compound, four known compounds — ferruginol, chamaecydin, 12-hydroxy-6,7-seco-abieta-8,l 1,13-triene-6,7-dial, and frans-communic acid — were isolated. All isolated compounds were subjected to an antifeedant bioassay against the pest insectSpodoptera litura. Results of the bioassay showed that chamaecydin and 12-hydroxy-6,7-seco-abieta-8,11,13-triene-6,7-dial had antifeedant activity.Part of this report was presented at 45th Annual Meeting of the Japan Wood Research Society, Tokyo, April 1995     

Individual variations in monoterpenes released from Cryptomeria japonica and Pinus thunbergii needles

Variations in the compositions of low boiling point (LBP) monoterpenes in needle samples from 99 sugi (Cryptomeria japonica) and 100 kuromatsu (Pinus thunbergii) trees were investigated using a headspace technique. Considerable variations in the proportions of monoterpenes were revealed in both species. In sugi, the proportions of sabinene and α-pinene in the total LBP monoterpenes, ranging from 8.8% to 73.3% and from 14.5% to 73.3%, respectively, showed enormous variations among nine monoterpenes. The proportions of 3-carene and limonene, ranging from 0.1% to 29.5% and from 0.2% to 20.4%, respectively, also showed very specific variations. In kuromatsu, the proportions of β-pinene and α-pinene in the total LBP monoterpenes, ranging from 26.5% to 66.3% and from 18.7% to 46.9%, respectively, showed considerable variations among ten monoterpenes. The proportions of myrcene and 1,8-cineole, ranging from 0.9% to 18.5% and from 0.8% to 12.3%, respectively, also showed specific variations. Part of this article was presented at the 50th Annual Meeting of the Japan Wood Research Society, Kyoto, April, 2000, and the 13th Annual Meeting of the Chugoku Shikoku Branch of the Japan Wood Research Society, Okayama, September 2001     

Stereochemistry and biosynthesis of (+)-lyoniresinol,a syringyl tetrahydronaphthalene lignan in <Emphasis Type="Italic">Lyonia ovalifolia</Emphasis> var. <Emphasis Type="Italic">elliptica</Emphasis> II: feeding experiments with <Superscript>14</Superscript>C labeled precursors

To clarify the biosynthetic pathway for syringyl lignans, especially syringyl tetrahydronaphthalene lignans and formation of the C2–C7′ linkage, production of (+)-lyoniresinol (LYR) and its predicted intermediates [syringaresinol (SYR), 5,5′-dimethoxylariciresinol (DMLR), and 5,5′-dimethoxysecoisolariciresinol (DMSLR)] in Lyonia ovalifolia var. elliptica was investigated by means of feeding experiments with radiolabeled precursors. Following individual administration of l-[U-¹⁴C]phenylalanine (Phe), [8-¹⁴C]sinapyl alcohol (SA), and [8,8′-¹⁴C]SYR to excised young shoots of L. ovalifolia and their subsequent metabolism, free [¹⁴C]lignans and [¹⁴C]lignan glycosides were extracted with methanol from stems and leaves and were divided into ethyl acetate-soluble fractions (lignans) and aqueous fractions (lignan glycosides), respectively. Using a combination of xylanase, cellulase, and β-glucosidase, the glycosides were hydrolyzed to liberate [¹⁴C]lignans as aglycones. l-[U-¹⁴C]Phe was incorporated into (+)-[¹⁴C]SYR [stem 0.38%, 8% enantiomeric excess (e.e.)], (−)-[¹⁴C]SYR (leaves 2.75%, 72% e.e.), (+)-[¹⁴C]DMLR (stem 0.07%, 18% e.e. and leaves 0.009%, 58% e.e.), (−)-[¹⁴C]DMSLR (stem 0.03%, 46% e.e. and leaves 0.05%, 20% e.e.), (+)-[¹⁴C]LYR (leaves 0.013%, 22% e.e.) and glycosides of (+)-[¹⁴C]LYR (stem 0.036%, 50% e.e.) in 24h. Based on the percent incorporation and enantiomeric composition of the lignans, the biosynthetic pathway of (8R,8′R)-(+)-LYR was proposed as follows: a nonselective dehydrogenative dimerization of sinapyl alcohol yields (±)-SYR, which is reduced with low specificity to give (8R,8′R)-(+)-DMLR. This is cyclized to directly give (+)-LYR as well as reduced again to (8R,8′R)-(−)-DMSLR. Although further transformation of (−)-DMSLR also leads to the formation of (+)-LYR, cyclization could be a main pathway for (+)-LYR biosynthesis. This report was presented at the IAWPS 2005 International Symposium on Wood Science and Technology, Yokohama, November 2005     

Biosynthesis of a syringyl 8-O-4′ neolignan in Eucommia ulmoides: formation of syringylglycerol-8-O-4′-(sinapyl alcohol) ether from sinapyl alcohol

To investigate the biosynthesis and stereochemistry of syringylglycerol-8-O-4′-(sinapyl alcohol) ether (SGSE), a syringyl 8-O-4′ neolignan, feeding experiments and enzyme assays using Eucommia ulmoides were carried out. Diastereoselective formation of erythro-SGSE was found. When [8-¹⁴C]sinapyl alcohol was administered to excised shoots of E. ulmoides, ¹⁴C was incorporated into free SGSE and SGSE glucosides. In stems, incorporation into (+)-erythro-[¹⁴C]SGSE (0.037%) with 9.1% enantiomeric excess (% e.e.) was found; incorporation into the threo isomer was not detectable. Erythro-[¹⁴C]SGSE glucosides (0.047%) dominated over threo forms (0.007%) with 74.0% diastereomeric excess (% d.e.); both diastereomers were levorotatory with 32.0% e.e. and 18.3% e.e., respectively. In leaves, higher incorporation into (−)-erythro-[¹⁴C]SGSE (0.500%, 15.9% e.e.) than into the threo isomer (0.206%, 7.4% e.e.) was observed (41.6% d.e.). (−)-Erythro-[¹⁴C]SGSE glucosides (1.692%, 25.0% e.e.) were produced at higher rates than threo isomers (0.177%, 16.4% e.e.) with 81.0% d.e. In incubations of a mixture of [8-¹⁴C]sinapyl and [8-¹⁴C]coniferyl alcohols with an insoluble enzyme preparation from stems of E. ulmoides, erythro-SGSE was preferentially produced. The highest % d.e. (82.8) was observed at 60 min with the (+)-erythro isomer (21.4% e.e.) and the (−)-threo form (4.3% e.e.).Part of this report was presented at the 52nd Annual Meeting of the Japan Wood Research Society, Gifu, April 2002, and the 47th Lignin Symposium, Fukuoka, October 2002     

Chemical constituents investigation of Daphne tangutica

A phytochemical study of an ethanol-soluble extract from the root barks of Daphne tangutica Maxim., a traditional Tibetan herb medicine, led to the isolation of 30 compounds, including eight daphnane diterpenes, nine coumarines, six lignans, five phenylpropanoid derivatives, β-sitosterol and p-hydroxy benzonate. Two compounds out of these isolates are new daphne diterpene analogs, and their structures were established as 1,2α-dihydro-5β-hydroxy-6α,7α-epoxy-resiniferonol-14-benzonate, and 1,2β-dihydro-5β-hydroxy-6α,7α-epoxy-resiniferonol-14-benzonate, respectively, on the basis of spectroscopic methods. Additionally, this is the first time that 13 known compounds have been isolated and identified from this traditional Tibetan medicinal plant.     

Heartwood extractives from the Amazonian trees <Emphasis Type="Italic">Dipteryx odorata,Hymenaea courbaril</Emphasis>, and <Emphasis Type="Italic">Astronium lecointei</Emphasis> and their antioxidant activities

Heartwood extracts from Amazonian trees cumaru-ferro (Dipteryx odorata), jatoba (Hymenaea courbaril), and guarita (Astronium lecointei) exhibit antioxidant activities comparable with that of α-tocopherol, a well-known antioxidant. This article reports the characterization of the antioxidant compounds in the extracts of the three heartwoods. Silica gel column chromatography of the cumaru-ferro EtOAc extract yielded (−)-(3R)-7,2′,3′-trihydroxy-4′-methoxyisoflavan and (+)-(3R)-8,2′,3′-trihydroxy-7,4′-dimethoxyisoflavan. Silica gel column chromatography followed by preparative high-performance liquid chromatography of the jatoba EtOAc extract yielded (−)-fisetinidol and (+)-trans-taxifolin. Chemical structures were assigned using electron-ionization mass spectrometry, ¹H and ¹³C nuclear magnetic resonance (NMR) spectroscopy including nuclear Overhauser effect spectroscopy (NOESY), as well as optical rotation and circular dichroism. Gas chromatography-mass spectrometry demonstrated that the isolated compounds were predominant in the EtOAc extracts. In the guarita EtOAc extract, catechin and gallic acid were identified by comparing their retention times and mass fragmentation patterns with those of authentic samples. Antioxidant activity determined by the 1,1-diphenyl-2-picrylhydrazyl assay demonstrated that all these compounds had activities comparable with that of α-tocopherol. Part of this report was presented at the 57th Annual Meeting of the Japan Wood Research Society, Hiroshima, August 2007     

Cultivation of <Emphasis Type="Italic">Pleurotus eryngii</Emphasis> on umbrella plant (<Emphasis Type="Italic">Cyperus alternifolius</Emphasis>) substrate

Mycelial growth and mushroom yields of three strains of Pleurotus eryngii produced on wheat bran-supplemented umbrella plant (Cyperus alternifolius) substrate were assessed using surface brightness, bromophenol blue color reactions, ergosterol and glucosamine contents, and water potential as indicators of strain performance. Mycelial growth was 31%–46% greater, depending on strain, on the umbrella plant substrate compared with the mushroom industry standard sugi (Cryptomeria japonica) substrate. Mushroom yields on the first flush were 20%–23% higher, depending on strain, on the plastic bottle-contained umbrella plant substrate. However, yields on the second break were lower from the umbrella plant substrate. Because many growers in Japan only harvest one flush, production of P. eryngii on umbrella plant substrate may offer commercial producers an alternative basal ingredient to diminishing supplies of sugi sawdust.Part of this report was presented at the 52nd Annual Meeting of the Japan Wood Research Society, Gifu, April 2002.     

Antimicrobial activity against <Emphasis Type="Italic">Streptococcus sobrinus</Emphasis> and glucosyltransferase inhibitory activity of taxifolin and some flavanonol rhamnosides from kempas (<Emphasis Type="Italic">Koompassia malaccensis</Emphasis>) extracts

Twenty plant materials collected from the islands of Java and Kalimantan in Indonesia were extracted with 50% aqueous ethanol (crude extract). The crude extracts were assayed for antimicrobial activities against Streptococcus sobrinus and for glucosyltransferase (GTase) inhibition. Fourteen extracts inhibited the growth of S. sobrinus by more than 50% and six extracts inhibited GTase activity by more than 50% at a concentration of 100 μg/ml. Koompassia malaccensis (kempas) extracts showed 90% depression of S. sobrinus growth and 80% inhibition of GTase activity at a concentration of 100 μg/ml. Kempas crude extracts were subjected to column chromatography using Sephadex LH-20 and then preparative high-performance liquid chromatography to isolate four compounds A, B, C, and D. These compounds were identified as taxifolin and the flavanonol rhamnoside isomers neoastilbin, astilbin, and isoastilbin, respectively, from ¹H and ¹³C nuclear magnetic resonance (NMR) spectra and other two-dimensional NMR techniques (COSY, HMBC, and HMQC). Each compound depressed the growth of S. sobrinus over a concentration range of 9.3242.7 μg/ml and showed GTase inhibitory activity with IC₅₀ values in the range 27.4–57.3 μg/ml. Taxifolin and flavanonol rhamnoside isomers isolated for the first time from kempas could be potent compounds for preventing dental caries. Part of this report was presented at the 57th Annual Meeting of the Japan Wood Research Society Conference, Hiroshima, 2007     

Extractives of <Emphasis Type="Italic">Quercus crispula</Emphasis> sapwood infected by the pathogenic fungi <Emphasis Type="Italic">Raffaelea quercivora</Emphasis> I: comparison of sapwood extractives from noninfected and infected samples

The extracts of Quercus crispula infected by the ambrosia fungus, Raffaelea quercivora, were investigated. Phenol and tannin analyses indicated that normal sapwood (NS) contained a considerable amount of hydrolysable tannins, while infected colored sapwood (IS) contained less hydrolysable tannins and more phenols than NS. In treating pentagalloyl glucose (PGG), which is a model compound of hydrolysable tannins, with a culture medium of R. quercivora, PGG was rapidly hydrolyzed to produce gallic acid. The resulting gallic acid decreased in concentration over the subsequent cultivation period eventually disappeared. Measuring tannase and laccase activities of the culture medium of R. quercivora, tannase activity increased gradually from the beginning, while laccase activity increased rapidly at 5 days of incubation and disappeared at 8 days. An oxidative product from gallic acid treated with laccase was isolated by preparative high performance liquid chromatography, and was identified as purprogallincarboxylic acid (PGCA) by nuclear magnetic resonance spectroscopy and electron-impact mass spectrometry. PGCA was present in a 70% aqueous acetone extract of IS, and showed slight growth inhibition against R. quercivora. Part of this study was presented at the 57th Annual Meeting of the Japan Wood Research Society, Hiroshima, Japan, 2007     

Screening of extracts of Japanese woods for melanin biosynthesis inhibition

The effects of 120 methanol extracts prepared from bark and heartwood of 69 types of Japanese wood on the melanin production of B16 melanoma cells were examined. The melanin content of B16 melanoma cells was determined spectrophotometrically at 405nm. The extracts were also examined for their effects on cell viability. We found that the methanol extracts of Fagus crenata (buna, wood, 100μg/ml), Sapium sebiferum (Nankinhaze, wood, bark, 10μg/ml), and Zelkova serrata (keyaki, wood, 10μg/ml) greatly inhibited the melanin production of B16 melanoma cells without significant cytotoxicity. However, these extracts did not inhibit tyrosinase activity at the concentration of 100μg/ml. These findings indicate that the depigmenting mechanism of these extracts involves the suppression of some pigmenting signals in stimulating melanogenesis rather than the inhibition of tyrosinase activity. Part of this study was presented at the 53rd Annual Meeting of the Japan Wood Research Society, Fukuoka, Japan, March 2003     

Stereochemistry and biosynthesis of 8-O-4′ neolignans in Eucommia ulmoides: diastereoselective formation of guaiacylglycerol-8-O-4′-(sinapyl alcohol) ether

Stereochemistry and biosynthesis of guaiacylglycerol-8-O-4′-(sinapyl alcohol) ether (GGSE), an 8-O-4′ neolignan, which consists of coniferyl and sinapyl alcohol moieties, in Eucommia ulmoides were investigated. Four 8-O-4′ neolignans, GGSE, syringylglycerol-8-O-4′-(coniferyl alcohol) ether (SGCE), guaiacylglycerol-8-O-4′-(coniferyl alcohol) ether (GGCE), and syringylglycerol-8-O-4′-(sinapyl alcohol) ether (SGSE), were synthesized. Their erythro and threo diastereomers were separated through acetonide derivatives, intermediates of the synthesis, and identified by means of nuclear magnetic resonance (NMR) spectroscopy. All of the erythro-acetonide derivatives have larger coupling constants (ca 9 Hz) for the C₇-H resonances than those of the threo ones (1.5–2 Hz). In the case of the four 8-O-4′ neolignans, the C₇-H coupling constants of the threo-isomers are not smaller than those of the erythro ones. GGSE isolated previously from this plant was identified as the erythro isomer by comparison of the ¹³C-NMR data with synthetic erythro-GGSE and threo-GGSE and the other 8-O-4′ neolignans mentioned as above. Administration of a mixture of [8-¹⁴C]coniferyl alcohol and [8-¹⁴C]sinapyl alcohol to excised shoots of E. ulmoides was carried out and the incorporation of ¹⁴C into erythro-[¹⁴C]GGSE was found to be higher than that in threo-[¹⁴C]GGSE. The occurrence of diastereoselective formation of erythro-GGSE by cross coupling of coniferyl and sinapyl alcohols is suggested.Part of this paper was presented at the 47th Lignin Symposium, Fukuoka, October 2002 and the 53rd Annual Meeting of the Japan Wood Research Society, Fukuoka, April 2003