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Metabolism of monoterpenes in mint (mentha) species.

Commercial mint ( MENTHA) species produce either C3-oxygenated or C6-oxygenated P-menthane monoterpene as the major essential oil components. The biosynthetic origin of both of these monoterpene families from the common precursor (-)-limonene is described. The metabolic turnover of monoterpenes occurs during late stages of plant development, and the pathway for the transport and catabolism of (-)-menthone in peppermint is reviewed as a model for this general process. The developmental regulation of monoterpene metabolism is briefly outlined.

Journal Article↗

Effect of Japanese mint (Mentha arvensis) oil as fumigant on nutritional quality of stored sorghum.

Japanese mint (Mentha arvensis) oil (JMO) can be used effectively as fumigant against Sitophilus oryzae in stored sorghum. The effect of JMO at a dose of 166 microliter/l of space on nutrient composition and protein quality was studied in infested and uninfested sorghum grains stored for 3 months. The results revealed non significant effect of JMO on gran moisture, total ash, crude fibre, crude fat, crude protein and fat acidity in infested and uninfested grains at the end of 3 months storage. The JMO treatment had small but significant effect on reducing and non-reducing sugars. The values of Protein Efficiency Ratio (PER) for uninfested JMO treated grains, infested JMO treated grains and for untreated control stored for 3 months were 1.11, 1.07 and 1.09, respectively against control casein diet for which it was 2.15.

Dietary Carbohydrates↗

Effect of Japanese mint (Mentha arvensis) oil as fumigant on stored sorghum: physical characteristics, sensory quality and germination.

The Japanese mint (Mentha arvensis) oil (JMO) was effective as fumigant against Sitophilus oryzae in sorghum (Sorghum bicolor). The observations on the effect of JMO treatment at a dose of 166.6 microliters/l of space to grains stored for 3 months in desiccators at 28 +/- 5 degrees C showed non significant (P approximately 0.05) effect on grain hardness, grain density and per cent water absorption. The cooking quality evaluated in terms of cooking time required for boiling of grains was also not significantly affected. The JMO treated samples of boiled sorghum scored significantly lower values for sensory quality characteristics viz. taste, aroma and overall acceptability compared to untreated samples. No effect of JMO on seed germination was observed. As sensory quality is lowered by use of JMO, the technique can only be recommended for seed sorghum preservation, not food.

Edible Grain↗

Functional expression of regiospecific cytochrome P450 limonene hydroxylases from mint (Mentha spp.) in Escherichia coli and saccharomyces cerevisiae.

The oxygenation pattern of the essential oil monoterpenes of commercial mint (Mentha) species is determined by regiospecific cytochrome P450-catalyzed hydroxylation of the common olefinic precursor (-)-4S-limonene. In spearmint (M. spicata), C6-allylic hydroxylation leads to (-)-trans-carveol and thence (-)-carvone, whereas in peppermint (M. x piperita), C3-allylic hydroxylation leads to (-)-trans-isopiperitenol and ultimately (-)-menthol. cDNAs encoding the C6-hydroxylase and C3-hydroxylase from spearmint and peppermint, respectively, were isolated by a combination of reverse genetic and homology-based cloning methods (S. Lupien, F. Karp, M. Wildung, and R. Croteau, Arch. Biochem. Biophys. 368, 181-192, 1999). Although both hydroxylase genes were confirmed by functional expression using the baculovirus-Spodoptera system, too little protein was available by this approach to permit detailed study of the structure-function relationships of these catalysts, especially the substrate binding determinants that underlie the regiochemistry and stereochemistry of the reactions. Therefore, heterologous overexpression systems based on Escherichia coli and Saccharomyces cerevisiae were developed to produce several N-terminally modified versions of the recombinant hydroxylases. Ancillary methods for the solubilization, purification, and reconstitution (with recombinant spearmint cytochrome P450 reductase) of the limonene hydroxylases were also devised, with which substrate binding behavior and precise regiochemistry and stereochemistry of product formation were determined.

Amino Acid Sequence↗

Effects of essential oil from mint (Mentha piperita) on Salmonella enteritidis and Listeria monocytogenes in model food systems at 4 degrees and 10 degrees C.

The effect of mint (Mentha piperita) essential oil (0.5, 1.0, 1.5 and 2.0%, v/w) on Salmonella enteritidis and Listeria monocytogenes in a culture medium and three model foods; tzatziki (pH 4.5), taramosalata (pH 5.0) and pâté (pH 6.8), inoculated at 10(7) cfu g-1, at 4 degrees and 10 degrees C for ca 1 week was studied. In the culture medium supplemented with the essential oil, no growth was observed over 2 d at 30 degrees C determined by a conductance method with a Malthus 2000 growth analyser. Salmonella enteritidis died in tzatziki in all treatments and declined in the other foods except for pâté at 10 degrees C as judged with viable counts. Listeria monocytogenes populations showed a declining trend towards the end of the storage period but was increased in pâté. Mint essential oil antibacterial action depended mainly on its concentration, food pH, composition, storage temperature and the nature of the micro-organism.

Animals↗

Effect of the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus fasciculatum on the essential oil yield related characters and nutrient acquisition in the crops of different cultivars of menthol mint (Mentha arvensis) under field conditions.

The effects of inoculation with vesicular-arbuscular mycorrhizal (VAM) fungus Glomusfasciculatum on the root colonization, growth, essential oil yield and nutrient acquisition of three cultivars of menthol mint (Mentha arvensis); Kalka, Shivalik and Gomti, were studied under field conditions. The VAM inoculation significantly increased the root colonization, plant height, fresh herbage and dry matter yield. oil content and oil yield as compared to non-inoculated cultivars. The effect of VAM inoculation on the root colonization, growth and yield of mint was more pronounced with the cv Shivalik than the cvs Kalka and Gomati, indicating Shivalik as a highly mycorrhizal dependent genotype. VAM inoculation significantly increased the uptake of N, P and K by shoot tissues of mint, but most markedly increased the uptake of P. The VAM-inoculated mint plants depleted the available N, P and K in the rhizosphere soil as compared to non-inoculated control plants, however the extent of nutrient depletion was greater for P than N and K. We conclude that the VAM inoculation could significantly increase the root colonization, growth, essential oil yield and nutrient acquisition of mint for obtaining economic production under field conditions.

Asteraceae↗

Regiospecific cytochrome P450 limonene hydroxylases from mint (Mentha) species: cDNA isolation, characterization, and functional expression of (-)-4S-limonene-3-hydroxylase and (-)-4S-limonene-6-hydroxylase.

The oxygenation pattern of the cyclic monoterpenoids of commercial mint (Mentha) species is determined by regiospecific cytochrome P450-catalyzed hydroxylation of the common olefinic precursor (-)-4S-limonene. In peppermint (Mentha x piperita), C3-allylic hydroxylation leads to (-)-trans-isopiperitenol, whereas in spearmint, C6-allylic hydroxylation leads to (-)-trans-carveol. The microsomal limonene-6-hydroxylase was purified from the oil glands of spearmint, and amino acid sequences from the homogeneous enzyme were used to design PCR primers with which a 500-bp amplicon was prepared. This nondegenerate probe was employed to screen a spearmint oil gland cDNA library from which the corresponding full-length cDNA was isolated and subsequently confirmed as the C6-hydroxylase by functional expression using the baculovirus-Spodoptera system. The probe was also utilized to isolate two closely related full-length cDNA species from a peppermint oil gland cDNA library which were confirmed as the limonene-3-hydroxylase by functional expression as before. Deduced sequence analysis of these regiospecific cytochrome P450 monooxygenases indicates that both enzymes bear a typical amino-terminal membrane anchor, consistent with the microsomal location of the native forms, exhibit calculated molecular weights of 56,149 (spearmint) and about 56,560 (peppermint), and are very similar in primary sequence (70% identity and 85% similarity). The availability of these regiochemically distinct, yet very closely related, recombinant hydroxylases and their corresponding genes provides a unique model system for understanding structure-function relationships in cytochrome P450 substrate binding and catalysis, and a means for transgenic manipulation of monoterpene biosynthetic pathways in plants.

Amino Acid Sequence↗

Hydroxylation of limonene enantiomers and analogs by recombinant (-)-limonene 3- and 6-hydroxylases from mint (Mentha) species: evidence for catalysis within sterically constrained active sites.

Limonene enantiomers and substrate analogs, including specifically fluorinated derivatives, were utilized to probe active site interactions with recombinant (-)-(4S)-limonene-3-hydroxylase (CYP71D13) and (-)-(4S)-limonene-6-hydroxylase (CYP71D18) from mint (Mentha) species. (-)-(4S)-Limonene is hydroxylated by both enzymes at the designated C3- and C6-allylic positions, with strict regio- and stereospecificity and without detectable allylic rearrangement, to give the corresponding products (-)-trans-isopiperitenol and (-)-trans-carveol. CYP71D13-catalyzed hydroxylation of (+)-(4R)-limonene also yields the corresponding trans-3-hydroxylated product ((+)-transisopiperitenol); however, the C6-hydroxylase converts (+)-(4R)-limonene to a completely different product profile dominated by the enantiopure cis-6-hydroxylated product (+)-cis-carveol along with several minor products, including both enantiomers of the trans-6-hydroxylated product ((+/-)-trans-carveol), indicating allylic rearrangement during catalysis. These results demonstrate that the regiospecificity and facial stereochemistry of oxygen insertion is dictated by the absolute configuration of the substrate. Fluorinated limonene analogs are also tightly bound by both enzymes and hydroxylated at the topologically congruent positions in spite of the polarizing effect of the fluorine atom on substrate reactivity. This strict retention of oxygenation geometry suggests a rigid substrate orientation imposed by multiple hydrophobic active site contacts. Structurally simplified substrate analogs are hydroxylated at slower rates and with substantial loss of regiospecificity, consistent with a loss of active site complementarity. Evaluation of the product profiles generated allowed assessment of the role of hydrophobic contacts in orienting the substrate relative to the activated oxygen species.

Catalytic Domain↗

Medicinal and aromatic plant materials as nitrification inhibitors for augmenting yield and nitrogen uptake of Japanese mint (Mentha arvensis L. Var. Piperascens).

Pot experiments were conducted to evaluate the relative performance of medicinal and aromatic plant materials and dicyandiamide (DCD) as nitrification inhibitors to regulate transformation of N from urea. Their effect on the efficiencies of use of N by Japanese mint (Mentha arvensis cv. Hy 77) was tested. Urea was coated with these materials viz., Mentha spicata, Artemisia annua or DCD at the rate of 5% (w/w) of fertilizer urea using an appropriate coating technique. Nimin (tetranortriterpenoids, an ethanol extract of neem (Azadirachta indica Juss) coating was done at the rate of 1% w/w of urea. Fertilizer nitrogen was applied at 100 and 200 mg kg(-1) soil. These natural coating materials significantly increased the herb and essential oil yields of the crop at both rates of fertilizer nitrogen compared to urea alone and were found to be as effective as DCD in retarding NO3- formation in soil. Herb yield increased by 6-81% when compared to uncoated urea. The increase in essential oil yield ranged between 3% and 68% due to coating. The effectiveness of the nitrification-inhibitor--coated urea, however, varied with the soils used and the rate of fertilizer nitrogen applied. The results suggest that the natural products could be potential nitrification inhibitors for increasing fertilizer N use efficiency.

Biological Availability↗

Analysis of Indian mint (Mentha spicata) for essential, trace and toxic elements and its antioxidant behaviour.

Mint, belonging to the genus Mentha in the family Labiatae (Lamiaceae) is pungent in taste with post digestive effects and hot potency. Ten samples of mint leaves, collected from four different locations in North-West parts of India (Roorkee, Dehradun, Baghpat and Uttarkashi) were analysed for seven minor (Al, Na, K, Ca, Cl, Mg, and P) and 20 trace (As, Au, Ba, Br, Co, Cr, Cs, Eu, Fe, Hf, Hg, La, Mn, Rb, Sb, Sc, Se, Sn, Th and Zn) elements by instrumental neutron activation analysis (INAA). Also Ni, Cu and Cd were determined by atomic absorption spectrophotometry (AAS). Samples along with reference materials (RMs) and synthetic primary standard were irradiated at approximately 10(13) n cm-2s-1 and its gamma-activity was measured using HPGe detector and MCA system. Most elements were found in widely varying amounts depending on the location, e.g. Na (0.21-0.86 mg/g), K (12.4-53.3 mg/g) and Ca (5.82-16.8 mg/g) whereas mean contents of other nutrient elements in mint were: Fe (108+/-22 microg/g), Mg (4.83+/-0.92 mg/g), Mn (53.5+/-9.6 microg/g), P (3.88+/-0.94 mg/g), Cu (16.9+/-1.8 microg/g), Zn (21.0+/-4.7 microg/g) and Se (0.18+/-0.03 microg/g). Further, DPPH free radical scavenging activity test in diethyl ether extract showed 100% activity at approximately 40 microg/L suggesting it to be antioxidant in accordance with literature reports.

Antioxidants↗

[Diversity of phytophagous and pathogens and their damage to mints (Mentha)].

UNLABELLED: The aim of this work was to estimate the phytopathologic condition of mints grown and self-growing in Kaunas Botanical Garden of Vytautas Magnus University and elsewhere in Lithuania; and to identify the specific constitution, the spread and damage of the main pests and disease agents. MATERIAL AND METHODS: Observations and examination were carried out with two local, four introduced mint species and with four hybrids in 1997-2003. We estimated the plant condition by identifying the intensity of the diseases and the abundance, spreading and damage of the pests. During the study we detected and identified the following pests--agents of diseases: Puccinia menthae, Erysiphe biocellata and Verticillium sp.; and phytophagous: Tetranychus urticae, Eupteryx atropunctata, Ovatus crataegarius, Polia persicariae, Agriotes lineatus, Chrysomela coerulans, Cassida viridis, Longitarsus lycopi. The most dangerous mint disease in Lithuania is rust; pests causing more harm are: Eupteryx atropunctata, Longitarsus lycopi. According to our data the least resistant species to most diseases and pests were Mentha arvensis and Mentha piperita, however separate samples of these species were exceptionally resistant. More resistant species was Mentha spicata and its hybrids.

Ecology↗

Optimization of water and nitrogen application to menthol mint (Mentha arvensis L.) through sugarcane trash mulch in a sandy loam soil of semi-arid subtropical climate.

Studies were carried out to optimize the use of water and nutrients by the crop with three moisture regimes [0.9, 1.2 and 1.5 irrigation water:cumulative pan evaporation (IW:CPE) ratios], two variables of organic mulch (control and sugarcane trash at 7 t/ha) and three levels of nitrogen (0, 100 and 200 kg/ha). Soil moisture regimes maintained at 1.2 IW:CPE ratio significantly increased the crop growth and herb and essential oil yields as compared with that of 0.9 IW:CPE ratio. The increase in herb yield due to 1.5 and 1.2 IW:CPE ratios was recorded to be 28.5% and 19%, respectively, over the irrigation given at 0.9 IW:CPE ratio, with the corresponding increase in essential oil yield to the extent of 23.5% and 15.5%. Interaction effect of moisture regimes and nitrogen rates indicated that increasing levels of irrigation at the highest level of N (200 kg/ha) improved essential oil yield of the crop. Application of N at 200 kg/ha in the mulched plots significantly enhanced the N uptake by the crop and essential oil yield over the control and 100 kg N/ha applied in the mulched/or unmulched plots and 200 kg N/ha applied in the unmulched plots. Application of organic mulch and nitrogen at 200 kg/ha improved the water use efficiency (WUE) in menthol mint crop. Higher moisture regimes maintained up to 1.2 IW:CPE ratio increased the WUE. The quality of essential oil in terms of its major constituent, menthol, improved slightly with 1.2 IW:CPE ratio as compared to 0.9 and 1.5 IW:CPE ratios at first and second harvests of the crop. It is recommended that menthol mint crop could be grown profitably by providing 16 irrigations, that is 80 cm water (based on 1.2 IW:CPE ratio) and nitrogen at 200 kg/ha in the sugarcane trash mulched plots, which could give a highest benefit:cost ratio from menthol mint cropping.

Agriculture↗

Demonstration that menthofuran synthase of mint (Mentha) is a cytochrome P450 monooxygenase: cloning, functional expression, and characterization of the responsible gene.

(+)-Menthofuran is an undesirable monoterpenoid component of peppermint (Mentha x piperita) essential oil that is derived from the alpha,beta-unsaturated ketone (+)-pulegone. Microsomal preparations, from the oil gland secretory cells of a high (+)-menthofuran-producing chemotype of Mentha pulegium, transform (+)-pulegone to (+)-menthofuran in the presence of NADPH and molecular oxygen, implying that menthofuran is synthesized by a mechanism analogous to that of mammalian liver cytochrome P450s involving the hydroxylation of the syn-methyl group of (+)-pulegone, spontaneous intramolecular cyclization to the hemiketal, and dehydration to the furan. An abundant cytochrome P450 clone from a peppermint oil gland cell cDNA library was functionally expressed in Saccharomyces cerevisiae and Escherichia coli and shown to encode the (+)-menthofuran synthase (i.e., (+)-pulegone-9-hydroxylase). The full-length cDNA contains 1479 nucleotides, and encodes a protein of 493 amino acid residues of molecular weight 55,360, which bears all of the anticipated primary structural elements of a cytochrome P450 and most closely resembles (35% identity) a cytochrome P450 monoterpene hydroxylase, (+)-limonene-3-hydroxylase, from the same source. The availability of this gene permits transgenic manipulation of peppermint to improve the quality of the derived essential oil.

Amino Acid Sequence↗

Mint virus X: a novel potexvirus associated with symptoms in 'Variegata' mint.

Mentha x gracilis 'Variegata', an ornamental mint clone first described about 200 years ago, exhibits virus-like vein banding symptoms. Double-stranded RNA and virion isolations revealed the presence of three viruses in a 'Variegata' plant. Cloning and sequencing disclosed that one of the viruses was a previously unidentified species with similarities to members of the Flexiviridae family, designated as Mint virus X (MVX). The complete nucleotide sequence of the virus was determined. Phylogenetic analysis divulged the close relationship of the virus with lily virus X and strawberry mild yellow edge virus, members of the Potexvirus genus. A reverse transcription-polymerase chain reaction protocol was developed and used for detection of MVX in other 'Variegata' plants. All clones tested, obtained from nurseries around the United States were infected with MVX, making the virus a possible causal agent of the variegated symptoms.

Genome, Viral↗

Influence of the leaf extract of Mentha arvensis Linn. (mint) on the survival of mice exposed to different doses of gamma radiation.

BACKGROUND: The aim of the present study was to evaluate the radioprotective effect of Mentha arvensis (mint) on the survival of mice exposed to various doses of whole-body gamma radiation. MATERIAL AND METHODS: The radioprotective effect of various doses (0, 2.5, 5, 10, 20, 40 and 80 mg/kg body weight) of chloroform extract of mint (Mentha arvensis Linn.) was studied in mice exposed to 10 Gy gamma radiation. RESULTS: The 10 mg/kg of mint extract was found to afford best protection as evidenced by the highest number of survivors in this group at 30 days post-irradiation, and further experiments were carried out using this dose of mint extract. The mice treated with 10 mg/kg body weight mint extract or oil were exposed to 6, 7, 8, 9 and 10 Gy of gamma radiation and observed for the induction of radiation-sickness and mortality up to 30 days post-irradiation. The mint extract pretreatment was found to reduce the severity of symptoms of radiation sickness and mortality at all exposure doses and a significant increase in the animal survival was observed when compared with the oil + irradiation group. All of the animals that were treated with 10 mg/kg mint extract and then exposed to 7 Gy irradiation were protected against the radiation-induced mortality when compared with the concurrent oil + irradiation group, in which 20% animals died by 30 days post-irradiation. The mint extract treatment protected the mice against the gastrointestinal death as well as bone marrow deaths. The DRF was found to be 1.2. The drug was non-toxic up to a dose of 1,000 mg/kg body weight, the highest drug dose that could be tested for acute toxicity. CONCLUSION: From our study it is clear that mint extract provides protection against the radiation-induced sickness and mortality and the optimum protective dose of 10 mg/kg is safe from the point of drug-induced toxicity.

Animals↗

Anti-HIV-1 activity of herbs in Labiatae.

The anti-HIV-1 activity of aromatic herbs in Labiatae was evaluated in vitro. Forty five extract from among 51 samples obtained from 46 herb species showed significant inhibitory effects against HIV-1 induced cytopathogenicity in MT-4 cells. In particular, the aqueous extracts of Melissa officinalis, a family of Mentha x piperita "grapefruit mint," Mentha x piperita var. crispa, Ocimum basilicum cv "cinnamon," Perilla frutescens var. crispa f. viridis, Prunella vulgaris subsp. asiatica and Satureja montana showed potent anti-HIV-1 activity (with an ED of 16 microg/ml). The active components in the extract samples were found to be water-soluble polar substances, not nonpolar compounds such as essential oils. In addition, these aqueous extracts inhibited giant cell formation in co-culture of Molt-4 cells with and without HIV-1 infection and showed inhibitory activity against HIV-1 reverse transcriptase.

Anti-HIV Agents↗