Kegg Pathway: Limonene and pinene degradation

Arch Biochem Biophys. 2008 Sep 1; 477(1): 121-30
Jones CG, Keeling CI, Ghisalberti EL, Barbour EL, Plummer JA, Bohlmann J

Sandalwood, Santalum album (Santalaceae) is a small hemi-parasitic tropical tree of great economic value. Sandalwood timber contains resins and essential oils, particularly the santalols, santalenes and dozens of other minor sesquiterpenoids. These sesquiterpenoids provide the unique sandalwood fragrance. The research described in this paper set out to identify genes involved in essential oil biosynthesis, particularly terpene synthases (TPS) in S. album, with the long-term aim of better understanding heartwood oil production. Degenerate TPS primers amplified two genomic TPS fragments from S. album, one of which enabled the isolation of two TPS cDNAs, SamonoTPS1 (1731bp) and SasesquiTPS1 (1680bp). Both translated protein sequences shared highest similarity with known TPS from grapevine (Vitis vinifera). Heterologous expression in Escherichia coli produced catalytically active proteins. SamonoTPS1 was identified as a monoterpene synthase which produced a mixture of (+)-alpha-terpineol and (-)-Limonene, along with small quantities of linalool, myrcene, (-)-alpha-pinene, (+)-sabinene and geraniol when assayed with geranyl diphosphate. Sesquiterpene synthase SasesquiTPS1 produced the monocyclic sesquiterpene alcohol germacrene D-4-ol and helminthogermacrene, when incubated with farnesyl diphosphate. Also present were alpha-bulnesene, gamma-muurolene, alpha- and beta-selinenes, as well as several other minor bicyclic compounds. Although these sesquiterpenes are present in only minute quantities in the distilled sandalwood oil, the genes and their encoded enzymes described here represent the first TPS isolated and characterised from a member of the Santalaceae plant family and they may enable the future discovery of additional TPS genes in sandalwood.

Metab Eng. 2008 May-Jul; 10(3-4): 166-77
Muñoz-Bertomeu J, Ros R, Arrillaga I, Segura J

We generated transgenic spike lavender (Lavandula latifolia) plants constitutively expressing the Limonene synthase (LS) gene from spearmint (Mentha spicata), encoding the LS enzyme that catalyzes the synthesis of Limonene from geranyl diphosphate. Overexpression of the LS transgene did not consistently affect monoterpene profile in pooled leaves or flowers from transgenic T(0) plants. Analyses from cohorts of leaves sampled at different developmental stages showed that essential oil accumulation in transgenic and control plants was higher in developing than in mature leaves. Furthermore, developing leaves of transgenic plants contained increased Limonene contents (more than 450% increase compared to controls) that correlated with the highest transcript accumulation of the LS gene. The levels of other monoterpene pathway components were also significantly altered. T(0) transgenic plants were grown for 2 years, self-pollinated, and the T(1) seeds obtained. The increased Limonene phenotype was maintained in the progenies that inherited the LS transgene.

Planta Med. 2008 Apr; 74(5): 515-20
Verma M, Singh SK, Bhushan S, Pal HC, Kitchlu S, Koul MK, Thappa RK, Saxena AK

The essential oil of Tanacetum gracile (Accession no. AT-01 termed AT-01 in the manuscript), a cold desert alpine highly aromatic herb, has 40 constituents including lavendulol (21.5 %), lavendulol acetate (1.7 %), alpha-pinene (11.2 %), 1,8-cineole (15.2 %), CIS-beta-ocimene (6.9 %), borneol (6.1 %), Limonene (5.1 %) and chamazulene (3.7 %). AT-01 was evaluated for its anticancer activity. It inhibited HL-60 cell proliferation with an IC (50) of 27 microg/mL. Furthermore, AT-01 induced apoptosis in human leukemia HL-60 cells as measured by several biological end points. AT-01 induced apoptotic body formation, enhanced annexinV-FITC binding of the cells, increased sub-G (0) DNA fraction, loss of mitochondrial membrane potential (Deltapsi (mt)) and release of cytochrome c from mitochondria, activated caspase-9 as well as caspase-3, and increased cleavage of PARP in HL-60 cells. Thus, AT-01 induced apoptosis through the mitochondrial dependent pathway in HL-60 cells.

J Microbiol Biotechnol. 2008 Mar; 18(3): 497-502
Lee JH, Yang HY, Lee HS, Hong SK

The essential oil from the cones of Pinus koraiensis was prepared after removing the seeds, and its chemical composition analyzed using gas chromatography-mass spectrometry (GC-MS). Hydrodistillation of the P. koraiensis cones yielded 1.07% (v/w) of essential oil, which was almost three times the amount of essential oil extracted from the needles of the same plant. Moreover, the antimicrobial activities of the oil against the growth of Gram-positive bacteria, Gram-negative bacteria, and fungi were evaluated using the agar disc diffusion method and broth microdilution method. Eighty-seven components, comprising about 96.8% of the total oil, were identified. The most abundant oil components were Limonene (27.90%), alpha-pinene (23.89%), beta-pinene (12.02%), 3-carene (4.95%), beta-myrcene (4.53%), isolongifolene (3.35%), (-)-bornyl acetate (2.02%), caryophyllene (1.71%), and camphene (1.54%). The essential oil was confirmed to have significant antimicrobial activities, especially against pathogenic fungal strains such as Candida glabrata YFCC 062 and Cryptococcus neoformans B 42419. Therefore, the present results indicate that the essential oil from the cones of Pinus koraiensis can be used in various ways as a nontoxic and environmentally friendly disinfectant.

Pest Manag Sci. 2008 Jul; 64(7): 736-47
Casado D, Gemeno C, Avilla J, Riba M

BACKGROUND: In recent years, many studies have been carried out on the behavioural and electrophysiological responses of Cydia pomonella (L.) to host volatile emissions, to find alternative attractants to the sex pheromone for pest monitoring. These studies have focused on apple and pear, and very little has been done on walnut. In the present work, the diurnal and seasonal variation in walnut volatile emissions and the electrophysiological response of C. pomonella have been studied. RESULTS: Ninety compounds were detected in walnut emissions, mainly monoterpenes and sesquiterpenes. The most abundant compound was beta-pinene, which, together with (Z)-3-hexenyl acetate, (E)-beta-ocimene, Limonene, germacrene D, 1,8-cineole, sabinene, (E)-beta-farnesene, (E)-beta-caryophyllene, beta-myrcene and beta-phellandrene, constituted between 81.9 and 90.5% of the total chromatographic area. Differences between seasonal periods were significant for 39 compounds, and between daytimes for 14 compounds. Discernible and consistent EAD responses were detected to 11 walnut-origin compounds, and confirmed with synthetics to seven of them. Except for alloocimene, pinocarvone and caryophyllene oxide, all these compounds are also emitted by apple. CONCLUSION: Walnut volatile emissions differ widely from apple ones, but both share many compounds that are EAD-active in C. pomonella. However, among EAD-active compounds there are three walnut-specific ones, which should be further tested in behavioural assays.

Plant Biol (Stuttg). 2008 Jan; 10(1): 163-9
Noe SM, Peñuelas J, Niinemets U

Research on biogenic volatile organic compound (BVOC) emissions has mainly focused on native species in natural ecosystems. However, much of the ozone and aerosol formation occurs in city atmospheres due to BVOC emissions by local urban vegetation. Plant composition of urban habitats is often dominated by non-native ornamental plant species, for which only limited data on BVOC emissions are available. To gain insight into the influence of ornamental vegetation on the urban atmospheric reactivity in Barcelona, Spain, we studied volatile isoprenoid emissions in 11 widespread ornamental tree species (three conifers and nine angiosperms). We found significant monoterpene emissions in all studied species, with normalized emission potentials (T=30 degrees C, photosynthetic photon flux density (PPFD)=1000 micromol x m(-2) x s(-1)) ranging between 0.2 to 110 microg x g(-1) (dry weight) h(-1). Depending on species, the emissions were dominated by alpha- and beta-pinene, myrcene, alpha- and beta-phellandrene, carene, Limonene and eucalyptol. These data demonstrate that ornamental plants may significantly contribute to the BVOC load in urban atmospheres and also underscore the importance of broadleaf angiosperms as significant monoterpene emitters.

Plant Biol (Stuttg). 2008 Jan; 10(1): 108-22
Rapparini F, Llusià J, Peñuelas J

Plant roots interact with a wide variety of rhizospheric microorganisms, including bacteria and the symbiontic arbuscular mycorrhizal (AM) fungi. The mycorrhizal symbiosis represents a series of complex feedbacks between plant and fungus regulated by their physiology and nutrition. Despite the widespread distribution and ecological significance of AM symbiosis, little is known about the potential of AM fungi to affect plant VOC metabolism. The purpose of this study was to investigate whether colonization of plant roots by AM fungi and associated soil microorganisms affects VOC emission and content of Artemisia annua L. plants (Asteraceae). Two inoculum types were evaluated: one consisted of only an arbuscular mycorrhizal (AM) fungus species (Glomus spp.), and the other was a mixture of different Glomus species and associated soil bacteria. Inoculated plants were compared with non-inoculated plants and with plants supplemented with extra phosphorus (P) to obtain plants of the same size as mycorrhizal plants, thus excluding potentially-confounding mycorrhizal effects on shoot growth. VOC emissions of Artemisia annua plants were analyzed by leaf cuvette sampling followed by off-line measurements with pre-concentration and gas chromatography mass spectrometry (GC-MS). Measurements of CO(2) and H(2)O exchanges were conducted simultaneously. Several volatile monoterpenes were identified and characterized from leaf emissions of Artemisia annua L. by GC-MS analysis. The main components identified belong to different monoterpene structures: alpha-pinene, beta-pinene, camphor, 1,8-cineole, Limonene, and artemisia ketone. A good correlation between monoterpene leaf concentration and leaf emission was found. Leaf extracts included also several sesquiterpenes. Total terpene content and emission was not affected by AM inoculation with or without bacteria, while emission of Limonene and artemisia ketone was stimulated by this treatment. No differences were found among treatments for single monoterpene content, while accumulation of specific sesquiterpenes in leaves was altered in mycorrhizal plants compared to control plants. Growth conditions seemed to have mainly contributed to the outcome of the symbiosis and influenced the magnitude of the plant response. These results highlight the importance of considering the below-ground interaction between plant and soil for estimating VOC emission rates and their ecological role at multitrophic levels.

Int J Food Microbiol. 2008 Feb 29; 122(1-2): 135-9
Rasooli I, Fakoor MH, Yadegarinia D, Gachkar L, Allameh A, Rezaei MB

Aflatoxin B1 (AFB1) is a highly toxic and carcinogenic metabolite produced by Aspergillus species on food and agricultural commodities. Natural products may regulate the cellular effects of aflatoxins and evidence suggests that aromatic organic compounds of spices can control the production of aflatoxins. With a view to controlling aflatoxin production, the essential oils from Rosmarinus officinalis and Trachyspermum copticum L. were obtained by hydrodistillation. Antifungal activities of the oils were studied with special reference to the inhibition of Aspergillus parasiticus growth and aflatoxin production. Minimal inhibitory (MIC) and minimal fungicidal (MFC) concentrations of the oils were determined. T. copticum L. oil showed a stronger inhibitory effect than R. officinalis on the growth of A. parasiticus. Aflatoxin production was inhibited at 450 ppm of both oils with that of R. officinalis being stronger inhibitor. The oils were analyzed by GC and GC/MS. The major components of R. officinalis and T. copticum L. oils were Piperitone (23.65%), alpha-pinene (14.94%), Limonene (14.89%), 1,8-Cineole (7.43%) and Thymol (37.2%), P-Cymene (32.3%), gamma-Terpinene (27.3%) respectively. It is concluded that the essential oils could be safely used as preservative materials on some kinds of foods to protect them from toxigenic fungal infections.

Nat Prod Res. 2008 Jan 10; 22(1): 31-6
Boehme AK, Noletto JA, Haber WA, Setzer WN

The leaf essential oils of Zanthoxylum rhoifolium and Zanthoxylum setulosum (Rutaceae) from Monteverde, Costa Rica have been obtained by hydrodistillation and analyzed by gas chromatography-mass spectrometry. The principal constituents of Z. rhoifolium leaf oil were germacrene D (14.6%), Limonene (12.5%), trans-2-hexenal (11.3%), beta-elemene (9.2%), 2-undecanone (9.2%), myrcene (7.9%), bicyclogermacrene (7.5%), and germacrene A (5.2%). The leaf oil of Z. setulosum was composed largely of beta-phellandrene (37.5%), beta-caryophyllene (13.7%), alpha-pinene (11.9%), germacrene D (10.9%), myrcene (5.9%), and nerolidol (5.4%). The essential oils were screened for in-vitro cytotoxic activity against Hep G2, MCF-7, and PC-3 human tumor cell lines; antibacterial activity against Bacillus cereus, Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli; and for Artemia salina (brine shrimp) lethality. Neither Z. rhoifolium nor Z. setulosum leaf oils exhibited cytotoxicity or antibacterial activity. Both oils showed activity against A. salina.

Arch Biochem Biophys. 2007 Dec 1; 468(1): 140-6
Karp F, Zhao Y, Santhamma B, Assink B, Coates RM, Croteau RB

The tightly coupled nature of the reaction sequence catalyzed by monoterpene synthases has prevented direct observation of the topologically required isomerization step leading from geranyl diphosphate to the enzyme-bound, tertiary allylic intermediate linalyl diphosphate, which then cyclizes to the various monoterpene skeletons. X-ray crystal structures of these enzymes complexed with suitable analogues of the substrate and intermediate could provide a clearer view of this universal, but cryptic, step of monoterpenoid cyclase catalysis. Toward this end, the functionally inert analogues 2-fluorogeranyl diphosphate, (+/-)-2-fluorolinalyl diphosphate, and (3R)- and (3S)-homolinalyl diphosphates (2,6-dimethyl-2-vinyl-5-heptenyl diphosphates) were prepared, and compared to the previously described substrate analogue 3-azageranyl diphosphate (3-aza-2,3-dihydrogeranyl diphosphate) as inhibitors and potential crystallization aids with two representative monoterpenoid cyclases, (-)-Limonene synthase and (+)-bornyl diphosphate synthase. Although these enantioselective synthases readily distinguished between (3R)- and (3S)-homolinalyl diphosphates, both of which were more effective inhibitors than was 3-azageranyl diphosphate, the fluorinated analogues proved to be the most potent competitive inhibitors and have recently yielded informative liganded structures with Limonene synthase.

Tree Physiol. 2007 Dec; 27(12): 1701-10
Woodward S, Bianchi S, Bodles WJ, Beckett L, Michelozzi M

Forty-one 2-year-old clones of Picea sitchensis (Bong.) Carr. from three full-sib families (14 clones from each of two families and 13 clones from a third family) were either wounded and inoculated with an isolate of Heterobasidion annosum (Fr.) Bref. or wounded without inoculation. Lesion lengths on the inner bark from the point of inoculation varied among clones 35 days after treatment. There was no relationship between lesion length and relatedness within families. Two clones (21342 and 25202) with the shortest lesions, tentatively designated as less susceptible to H. annosum, and two clones (21176 and 27166) with the longest lesions, designated more susceptible, were selected for comparison of host anatomical and chemical responses to infection. The position and structure of the ligno-suberized boundary zone (LSZ) in the bark of the clones suggested that the less susceptible clones formed thicker layers of suberized cells in the LSZ following wounding plus inoculation. No LSZ was observed in two ramets of the more susceptible Clone 27166 following wounding and inoculation with H. annosum. Compared with more susceptible genotypes, clones of P. sitchensis with low susceptibility to H. annosum had high relative proportions of (+)-alpha-pinene, (-)-beta-pinene and one unidentified terpene constituent (Unknown-15) in cortical resin sampled 25 cm from the lesions. In contrast, more susceptible clones had higher relative proportions of (-)-Limonene, Unknown-16, Unknown-18 and Unknown-19. In the secondary resin produced in bark tissues surrounding the lesions, proportions of several monoterpene constituents varied; these changes included a decrease in the relative amount of beta-phellandrene and corresponding small increases in some minor constituents. The concentrations of the monoterpenes, except a few minor constituents, increased in the infected tissues. Wounding plus inoculation with H. annosum resulted in varied monoterpene responses, with distinct differences between less susceptible and more susceptible clones. In less susceptible clones, Unknown-19 increased following wounding plus inoculation, whereas in more susceptible clones, concentrations of delta-3-carene and Unknown-13 and Unknown-16 increased. Differences in both constitutive and induced resin monoterpene profiles may provide useful markers for resistance to H. annosum in selection and breeding programs.

Environ Sci Technol. 2007 Sep 1; 41(17): 6129-36
Heaton KJ, Dreyfus MA, Wang S, Johnston MV

The formation of secondary organic aerosol (SOA) by reaction of ozone with monoterpenes (beta-pinene, delta3-carene, Limonene, and sabinene) was studied on a short time scale of 3-22 s with a flow tube reactor. Online chemical analysis was performed with the Photoionization Aerosol Mass Spectrometer (PIAMS) to obtain molecular composition and the Nanoaerosol Mass Spectrometer (NAMS) to obtain elemental composition. Molecular composition data showed that dimers and higher order oligomers are formed within seconds after the onset of reaction, indicating that there is no intrinsic kinetic barrier to oligomer formation. Because oligomer formation is fast, it is unlikely that a large number of steps are involved in their formation. Therefore, ion distributions in the PIAMS spectra were interpreted through reactions of intermediates postulated in previous studies with monomer end products or other intermediates. Based on ion signal intensities in the mass spectra, organic peroxides appear to comprise a greater fraction of the aerosol than secondary ozonides. This conclusion is supported by elemental composition data from NAMS that gave C:O ratios in the 2.2-2.7 range.

Arch Biochem Biophys. 2007 Sep 15; 465(2): 417-29
Landmann C, Fink B, Festner M, Dregus M, Engel KH, Schwab W

The essential oil of lavender (Lavandula angustifolia) is mainly composed of mono- and sesquiterpenes. Using a homology-based PCR strategy, two monoterpene synthases (LaLIMS and LaLINS) and one sesquiterpene synthase (LaBERS) were cloned from lavender leaves and flowers. LaLIMS catalyzed the formation of (R)-(+)-Limonene, terpinolene, (1R,5S)-(+)-camphene, (1R,5R)-(+)-alpha-pinene, beta-myrcene and traces of alpha-phellandrene. The proportions of these products changed significantly when Mn(2+) was supplied as the cofactor instead of Mg(2+). The second enzyme LaLINS produced exclusively (R)-(-)-linalool, the main component of lavender essential oil. LaBERS transformed farnesyl diphosphate and represents the first reported trans-alpha-bergamotene synthase. It accepted geranyl diphosphate with higher affinity than farnesyl diphosphate and also produced monoterpenes, albeit at low rates. LaBERS is probably derived from a parental monoterpene synthase by the loss of the plastidial signal peptide and by broadening its substrate acceptance spectrum. The identification and description of the first terpene synthases from L. angustifolia forms the basis for the biotechnological modification of essential oil composition in lavender.

Plant Mol Biol. 2007 Sep; 65(1-2): 107-24
Roeder S, Hartmann AM, Effmert U, Piechulla B

The white flowers of N. suaveolens emit a complex bouquet of fragrance volatiles. The dominant compounds are benzenoids (e.g. methyl benzoate, methyl salicylate, benzyl benzoate and benzyl salicylate), monoterpenes (1,8-cineole, Limonene, sabinene, E-beta-ocimene, beta-beta-myrcene, alpha- and beta-pinene and alpha-terpineole) and sesquiterpenes (e.g. caryophyllene), which are all emitted at higher levels during the night. Here, we show that the simultaneous nocturnal emission of most monoterpenes is realized by a single floral-specific multi-product enzyme (1,8-cineole synthase, CIN), which synthesizes the monoterpenes of the "cineole cassette". Interestingly, N. suaveolens is the only known taxon of the Suaveolentes section to have a flower emitting "cineole cassette of monoterpenes" which is otherwise typical for the Alatae section. Gene sequence analysis of CIN has revealed the highest similarities to other angiosperm monoterpene synthases from Vitis vinifera, Quercus ilex, Citrus unshiu and C. limon, which cluster in the same branch of the terpene synthase B subfamily. However, based on its synthesized products, N. suaveolens CIN shares similarity with enzymes of the Arabidopsis thaliana root and Salvia officinalis leaf. The N. suaveolens CIN gene is only expressed in the stigma/style tissue and petals. Thin sections of petals present the enzyme primarily in the adaxial and abaxial epidermis; this facilitates the comprehensive emission of volatiles in all spacial directions. The oscillation of monoterpene emission is a consequence of the regulation of the CIN gene by the circadian clock, with oscillations occurring at the level of transcript and protein accumulations and of enzyme activity. Light/dark or dark/light transition signals synchronize the slow-running endogenous clock. Two strategies for synchronized scent emission have been established in N. suaveolens flowers: (i) the synthesis of volatile organic compounds by a multi-product enzyme and (ii) the coordination of biosynthetic pathways by a circadian clock.

FEMS Microbiol Lett. 2007 Apr; 269(2): 323-30
Rhodes AH, Owen SM, Semple KT

It has been suggested that monoterpenes emitted within the soil profile, either by roots or by decaying biomass, may enhance the biodegradation of organic pollutants. The aim of this study was to evaluate the effect of biogenic volatile organic compounds (VOCs) on the catabolism of 2,4-dichlorophenol in soils. Soils were collected from areas surrounding monoterpene (woodland) and nonmonoterpene (grassland)-emitting vegetation types. Soils were spiked with [UL-14C] 2,4-dichlorophenol at 10 mg kg(-1) and amended with alpha-pinene, p-cymene or a mix of monoterpenes (alpha-pinene, Limonene and p-cymene in 1:1:1 ratio). The effects of monoterpene addition on the catabolism of [UL-14C] 2,4-dichlorophenol to 14CO2 by indigenous soil microbial communities were assessed in freshly spiked and 4-week-aged soils. It was found that aged woodland soils exhibited a higher level of [UL-14C] 2,4-dichlorophenol degradation, which was subsequently enhanced by the addition of monoterpenes (P<0.001), with the VOC mix and alpha-pinene amendments showing increased [UL-14C] 2,4-dichlorophenol catabolism. This study supports claims that the addition of biogenic VOCs to soils enhances the degradation of xenobiotic contaminants.

Proc Natl Acad Sci U S A. 2007 Mar 27; 104(13): 5360-5
Hyatt DC, Youn B, Zhao Y, Santhamma B, Coates RM, Croteau RB, Kang C

The crystal structure of (4S)-Limonene synthase from Mentha spic ata, a metal ion-dependent monoterpene cyclase that catalyzes the coupled isomerization and cyclization of geranyl diphosphate, is reported at 2.7-A; resolution in two forms liganded to the substrate and intermediate analogs, 2-fluorogeranyl diphosphate and 2-fluorolinalyl diphosphate, respectively. The implications of these findings are described for domain interactions in the homodimer and for changes in diphosphate-metal ion coordination and substrate binding conformation in the course of the multistep reaction.

J Chem Ecol. 2007 Apr; 33(4): 749-65
Füssel U, Dötterl S, Jürgens A, Aas G

The floral scent composition of 32 European and two Asian Salix L. species (Salicaceae) was analyzed. Intra- and interspecific variation was compared for a subset of 8 species. All Salix species are dioecious and floral scent was collected from both male and female individuals by using a dynamic headspace MicroSPE method, and analyzed by GC-MS. A total of 48 compounds were detected, most of them being isoprenoids and benzenoids. Commonly occurring compounds included trans-beta-ocimene, cis-beta-ocimene, benzaldehyde, D-Limonene, alpha-pinene, cis-3-hexenyl aceatate, linalool, 1,4-dimethoxybenzene, and beta-pinene. Two compounds, 1,4-dimethoxybenzene and trans-beta-ocimene, were responsible for most of the interspecific variation. In a subset of eight extensively sampled species, six had a characteristic floral scent composition; half of the pairwise species comparisons confirmed significant differences. In three of these eight species, intraspecific variability could be explained by sex differences. Variation in Salix floral scent may provide specific signals that guide pollinators and thus contribute to the reproductive isolation of compatible and cooccurring species.

J Chromatogr A. 2007 Jun 8; 1152(1-2): 262-7
Yang Y, Kayan B, Bozer N, Pate B, Baker C, Gizir AM

In the first part of this study, the stability of five terpenes (alpha-pinene, Limonene, camphor, citronellol, and carvacrol) under subcritical water conditions was investigated. The stability studies were carried out at four different temperatures (100, 150, 200, and 250 degrees C) with two different heating times (30 and 300 min). When water temperature was increased, the degradation of terpenes became more serious. Prolonged exposure time to each heating temperature also caused decreased terpene stability. The terpene recoveries were determined by conducting subcritical water extraction of sand spiked with terpenes. The recoveries are typically around 70 to 80% for extractions at 100 degrees C. Terpene recoveries were decreased with increasing water temperature due to poorer stability of terpenes. After the degradation and recovery studies, basil and oregano leaves were extracted using water at both 100 and 150 degrees C. The concentrations of each individual terpene in the water extract generally ranged from trace quantity to 65 microg terpene/g herb. However, the concentration of carvacrol in the oregano-water extract at 150 degrees C was found to be as high as 4270 microg carvacrol/g oregano.

Appl Biochem Biotechnol. 2006; 129-132: 1077-88
Jeong GT, Park DH, Lee GY, Cha JM

Biofiltration is a biological process which is considered to be one of the more successful examples of biotechnological applications to environmental engineering, and is most commonly used in the removal of odoriferous compounds. In this study, we have attempted to assess the efficiency with which both single and complex odoriferous compounds could be removed, using one- or two-stage biofiltration systems. The tested single odor gases, Limonene, alpha-pinene, and iso-butyl alcohol, were separately evaluated in the biofilters. Both Limonene and alpha-pinene were removed by 90% or more EC (elimination capacity), 364 g/m3/h and 321 g/m3/h, respectively, at an input concentration of 50 ppm and a retention time of 30 s. The iso-butyl alcohol was maintained with an effective removal yield of more than 90% (EC 375 g/m3/h) at an input concentration of 100 ppm. The complex gas removal scheme was applied with a 200 ppm inlet concentration of ethanol, 70 ppm of acetaldehyde, and 70 ppm of toluene with residence time of 45 s in a one- or two-stage biofiltration system. The removal yield of toluene was determined to be lower than that of the other gases in the one-stage biofilter. Otherwise, the complex gases were sufficiently eliminated by the two-stage biofiltration system.

Nutr Neurosci. 2006 Feb-Apr; 9(1-2): 73-80
Fukumoto S, Sawasaki E, Okuyama S, Miyake Y, Yokogoshi H

Citrus essential oils have been utilized widely in traditional medicine, and there are various reports of actions such as effects on behavior and effects on pain stimulation response due to exposure. However, there are no reports concerning effects on neurotransmitters after ingestion, and uptake within the brain. We used brain tissue slices to investigate the effect of compounds in lemon essential oil on monoamine release. We investigated R-Limonene, gamma-terpinene and citral, major components of lemon essential oil; S-Limonene, an isomer of R-Limonene and metabolites of these compounds. The effect of each compound on monoamine release from brain tissue slices was found to be dose-dependent. R-Limonene and its S-isomer demonstrated differences with regard to monoamine release from brain tissue. S-Limonene and its metabolites were found to have a stronger effect than the R-isomer. Limonene metabolites taken up in vivo were also found to have a stronger effect on monoamine release than both the R-form and the S-form. In an investigation of dopamine release using stratum slices, terpinene and pinene demonstrated no clear differences in activity attributable to isomers. However, rho-cymene, a gamma-terpinene metabolite, was found to have a stronger effect than gamma-terpinene. These results suggest that the metabolites of these monoterpene compounds contained in citrus essential oils have a stronger effect on monoamine release from brain tissue than the monoterpene compounds themselves.