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PubMed1996

Bell pepper fruit fatty acid hydroperoxide lyase is a cytochrome P450 (CYP74B).

Fatty acid hydroperoxide lyases cleave a C-C bond adjacent to a hydroperoxide group in lipoxygenase derived lipid hydroperoxides to form short-chain aldehydes and oxo-acids. Previously, we showed that fatty acid hydroperoxide lyase from bell pepper fruits is a heme protein whose spectrophotometric properties greatly resemble a cytochrome P450. In order to ascertain the relationship of it to the P450 gene family, we have cloned cDNA encoding fatty acid hydroperoxide lyase from immature bell pepper fruits. The cDNA encodes 480 amino acids, and shares homology with P450s mostly at the C terminus. The heme binding cysteine is recognizable at position 441. The most closely related P450 is allene oxide synthase (CYP74A), with which it has 40% identity. It qualifies the lyase as a member of a new P450 subfamily, CYP74B. From this finding, the enzyme is thought to be a novel member of P450 specialized for the metabolism of lipid peroxides.

Aldehyde-Lyases↗

PubMed1994

Rapid detection and differentiation of tobamoviruses infecting L-resistant genotypes of pepper by RT-PCR and restriction analysis.

A procedure involving reverse transcription followed by polymerase chain reaction (RT-PCR) was developed for typing pathotypes of the tobamoviruses infecting the L-resistant genotypes of pepper. The method provides a much simpler alternative to the bioassay tests for the different Capsicum spp. genotypes previously used. Discrimination between the two pathotypes, P1,2 and P1,2,3, which cannot be differentiated by serological means, was achieved by restriction enzyme analysis of the PCR products. The assay also detects and distinguishes both pathotypes in a single mixed-infected plant. The procedure should be useful for the diagnosis and control of the disease and helpful to breeders and biotechnologists when producing and evaluating resistance in pepper plants.

Base Sequence↗

PubMed1996

Capsaicin in hot chili pepper: carcinogen, co-carcinogen or anticarcinogen?

Capsaicin (trans-8-methyl-N-vanillyl-6-nonenamide) is a major pungent ingredient of the Capsicum fruits such as hot green and red peppers. Besides its use as a food additive in various spicy cuisines, capsaicin is currently utilized for therapeutic purposes to treat various peripheral painful conditions such as rheumatoid arthritis and diabetic neuropathy. Considering consumption of capsaicin as a food additive and its current medicinal application in humans, correct evaluation and precise assessment of any harmful effects of this compound are essential from the public health standpoint. Numerous investigations have been conducted to determine the potential mutagenic and carcinogenic activity of capsaicin and chili pepper, but results are discordant. This review briefly examines findings in the literature of studies testing mutagenicity and tumorigenicity of capsaicin and presents a possible mechanistic basis for the dual effects exerted by the compound.

Animals↗

PubMed2005

Identification of a CaRAV1 possessing an AP2/ERF and B3 DNA-binding domain from pepper leaves infected with Xanthomonas axonopodis pv. glycines 8ra by differential display.

We isolated a cDNA clone, CaRAV1, which exhibited significant similarity to those of Arabidopsis RAV proteins containing AP2/ERF and B3-like DNA-binding domains. CaRAV1 expression was rapidly and specifically induced in both host and non-host resistant responses against bacterial pathogens in the chili pepper plant. CaRAV1 also strongly increased following salicylic acid and ethephon treatments, whereas methyl-jasmonate only had mild effects. Furthermore, CaRAV1 transcript levels were also investigated in response to ABA and abiotic stress. No significant CaRAV1 expression was evident following ABA, mannitol, or cold treatments. These observations collectively provide initial evidence that the pepper RAV transcription factor homolog may function in plant defense responses.

Acetates↗

PubMed2004

The CaPRP1 gene encoding a putative proline-rich glycoprotein is highly expressed in rapidly elongating early roots and leaves in hot pepper (Capsicum annuum L. cv. Pukang).

Most of the proline-rich cell wall glycoprotein genes isolated from higher plants are preferentially expressed in the transmitting tissues of the flower organ. In conducting expressed sequence tag (EST) analysis, which was prepared from 5-day-old early roots of hot pepper (Capsicum annuum L. cv. Pukang), we identified a cDNA clone, pCaPRP1, encoding a putative cell wall proline-rich glycoprotein. CaPRP1 (Mr=28 kDa, pI=9.98) was most closely related to Nicotiana alata NaPRP4 (71%), while most distantly related to soybean PvPRP (37%). The predicted primary structure of CaPRP1 contains a putative N-terminal signal peptide, six repeats of the Lys-Pro-Pro tripeptide, four repeats of a five-amino acid sequence [Pro-(Ser/The)-Pro-Pro-Pro] and one potential N-glycosylation site (Asn-Asn-Ser). In contrast to most proline-rich cell wall glycoprotein genes, CaPRP1 was highly expressed in rapidly elongating very early roots and young leaves as well as developing flower tissues. Although the physiological function of CaPRP1 is not yet clear, there are several possibilities for its role in cell expansion and elongation during early development of hot pepper plants.

Amino Acid Sequence↗

PubMed2006

In vivo binding of hot pepper bZIP transcription factor CabZIP1 to the G-box region of pathogenesis-related protein 1 promoter.

We find that salicylic acid and ethephon treatment in hot pepper increases the expression of a putative basic/leucine zipper (bZIP) transcription factor gene, CabZIP1. CabZIP1 mRNA is expressed ubiquitously in various organs. The green fluorescent protein-fused transcription factor, CabZIP1::GFP, can be specifically localized to the nucleus, an action that is consistent with the presence of a nuclear localization signal in its protein sequence. Transient overexpression of the CabZIP1 transcription factor results in an increase in PR-1 transcripts level in Nicotiana benthamiana leaves. Using chromatin immunoprecipitation, we demonstrate that CabZIP1 binds to the G-box elements in native promoter of the hot pepper pathogenesis-related protein 1 (CaPR-1) gene in vivo. Taken together, our results suggest that CabZIP1 plays a role as a transcriptional regulator of the CaPR-1 gene.

Amino Acid Sequence↗

PubMed2004

Investigation of acute toxicity of fenitrothion on peppered corydoras (Corydoras paleatus) (Jenyns, 1842).

Fenitrothion, as an organophosphothionate insecticide, is a contact insecticide and selective acaricide, also used as a vector control agent for malaria in public health programs. A 96 h LC50 value of fenitrothion, a potential toxic pollutant contaminating aquatic ecosystems, was determined on the adult peppered corydoras (Corydoras paleatus). The experiments were repeated three times. The static test method of acute toxicity test was used. Water temperature was regulated at 23 +/- 1 degrees C. In addition, behavioral changes at each fenitrothion concentration were observed for the individual fish. Data obtained from acute toxicity tests were evaluated using the Probit Analysis Statistical Method. The 96 h LC50 value for peppered corydoras was estimated as 3.51 mg/l.

Animals↗

PubMed2004

Effect of red pepper Capsicum annuum var. conoides and garlic Allium sativum on plasma lipid levels and cecal microflora in mice fed beef tallow.

Antihyperlipidemia or hypocholesterolaemic and antibacterial activities of red hot pepper and garlic are well known. To determine the effect of the dietary spices ingested to suppress blood lipids on the intestinal condition, we examined plasma lipid levels and cecal microflora in mice that were fed diets containing 19% (w/w) beef tallow and 2% red pepper Capsicum annuum var. conoides 'Takanotume' (RP) or garlic Allium sativum 'White' (GP) for 4-weeks. Plasma triacylglyceride level was suppressed by the spices. RP lowered cecal bacteroidaceae, a predominant bacterial group (from 9.4 to 9.0 log CFU/g), bifidobacteria (from 8.7 to 7.6 log CFU/g), and staphylococci. Although GP increased the cecal weight including their contents, significant differences were not shown in the cecal microflora. These results suggest that RP can affect the intestinal condition and host health through the disturbance of intestinal microflora.

Animals↗

PubMed2005

Activation of pepper basic PR-1 gene promoter during defense signaling to pathogen, abiotic and environmental stresses.

The basic PR-1 gene, CABPR1, accumulates in pepper leaf tissues during pathogen infection as well as after ethylene treatment. We isolated and functionally characterized the CABPR1 promoter region in tobacco leaves to identify the cis-acting regulatory sequences that are involved in CABPR1 gene expression. Constructs harboring the 5'-serially deleted CABPR1 promoter, which was fused to the beta-glucuronidase (GUS) gene, were evaluated for their promoter activity in the tobacco leaves. The CABPR1 promoter of 1670 bp in size was locally or systemically induced during a compatible interaction with Pseudomonas syringae pv. tabaci. The CABPR1 promoter also was differentially activated by treatment with ethylene, salicylic acid, nitric oxide, high salinity, drought and low temperature. The expression of the pepper transcription factors, CAZFP1 and CARAV1, activated the CABPR1 promoter. Analyses of a series of 5'-deletions of the CABPR1 promoter indicated that novel cis-acting elements essential for induction by pathogen and abiotic elicitors are localized in the region between -1670 bp and -1466 bp upstream from the translation start site. These results suggest that CABPR1 promoter is essential for regulating CABPR1 gene expression in response to pathogen, abiotic and environmental stresses, possibly by transactivating the CAZFP1 and CARAV1 transcription factors.

Base Sequence↗

PubMed2005

Composting of a solid olive-mill by-product ("alperujo") and the potential of the resulting compost for cultivating pepper under commercial conditions.

A pollutant solid material called "alperujo" (AL), which is the main by-product from the Spanish olive oil industry, was composted with a cotton waste as bulking agent, and the compost obtained (ALC) was compared with a cattle manure (CM) and a sewage sludge compost (SSC) for use as organic amendment on a calcareous soil. The experiment was conducted with a commercial pepper crop in a greenhouse using fertigation. Composting AL involved a relatively low level of organic matter biodegradation, an increase in pH and clear decreases in the C/N and the fat, water-soluble organic carbon and phenol contents. The resulting compost, which was rich in organic matter and free of phytotoxicity, had a high potassium and organic nitrogen content but was low in phosphorus and micronutrients. The marketable yields of pepper obtained with all three organic amendments were similar, thus confirming the composting performance of the raw AL. When CM and SSC were used for soil amendment, the soil organic matter content was significantly reduced after cultivation, while it remained almost unchanged in the ALC-amended plots.

Agriculture↗

PubMed2000

Clinical effects of oleoresin capsicum (pepper spray) on the human cornea and conjunctiva.

OBJECTIVE: To evaluate the effects of oleoresin capsicum (OC) on the human cornea and conjunctiva and to test the effectiveness of topical anesthetics for relief of pain. DESIGN: Prospective, randomized clinical trial. METHODS: Forty-seven subjects were examined before and at 10 minutes and 1 hour after exposure to pepper spray during a training exercise. Eleven subjects were reexamined at 1 week after exposure. A short, subjective questionnaire was given asking subjects to rate their pain, blurring of vision, and tearing. After exposure, subjects were randomly given a placebo, a topical nonsteroidal antiinflammatory agent, or a topical anesthetic. MAIN OUTCOME MEASURES: Visual acuity and corneal sensitivity with a Cochet-Bonnet aesthesiometer (scale of 1-6 cm) was measured and the eyes were examined with a portable slit lamp using fluorescein. Symptoms of pain, blurring of vision, and tearing were recorded in a ranking of 0 to 10. RESULTS: Visual acuity was unaffected by exposure to pepper spray. Corneal sensitivity was reduced from a pretest mean of 5.7 cm to a posttest mean of 0.6 cm 10 minutes after exposure. At 1 hour, the mean corneal sensitivity had recovered to 2.9 cm. Twenty-one percent of eyes had evidence of punctate epithelial erosions, but no corneal abrasions were found. All subjects reported significant pain, blurring of vision, and tearing at 10 minutes that was much improved by 1 hour. Topical flurbiprofen 0.03% improved symptoms in two of 11 subjects, whereas topical proparacaine hydrochloride 0.5% improved symptoms in 16 of 29 eyes. At 1 week after exposure, corneal sensation returned to baseline, and no corneal abnormalities were noted. CONCLUSIONS: The predominant symptom after exposure to OC was pain. Topical flurbiprofen was not helpful in reducing symptoms of exposure, whereas topical proparacaine was effective in relieving pain in most subjects. Corneal sensitivity was dramatically reduced at 10 minutes after exposure and was improved after 1 hour. At 1 week, corneal sensation had returned to normal, as had slit-lamp appearance on all subjects examined.

Adult↗

PubMed2003

Calcium restores prepenetration morphogenesis abolished by polyamines in Colletotrichum gloeosporioides infecting red pepper.

Colletotrichum gloeosporioides forms a specialized infection structure, an appressorium, to infect its host, red pepper. Polyamines (putrescine, spermidine, and spermine) as well as S-adenosyl methionine inhibitor, methylglyoxal-bis-guanyl hydrazone (MGBG), impaired conidial germination and appressorium formation of C. gloeosporioides. Curtailment of cell differentiation by polyamines and MGBG was more evident in conidial germination than in appressorium development. Exogenous addition of calcium restored conidial germination and appressorium formation and expression of calmodulin-encoding gene (CgCaM) inhibited by polyamines. Taken together, proper regulation of intracellular polyamine concentration is indispensable for conidial germination and appressorium formation, and involved in Ca(2+)/calmodulin-dependent signaling pathways of C. gloeosporioides infecting red pepper.

Calcium↗

PubMed2002

CYP3A4 inhibitory activity of new bisalkaloids, dipiperamides D and E, and cognates from white pepper.

Two new bisalkaloids, dipiperamides D and E, were isolated as inhibitors of a drug metabolizing enzyme cytochrome P450 (CYP) 3A4 from the white pepper, Piper nigrum. Their structures were elucidated by spectroscopic methods. Dipiperamides D and E showed potent CYP3A4 inhibition with IC(50) values of 0.79 and 0.12 microM, respectively, and other metabolites from the pepper were moderately active or inactive.

Alkaloids↗

PubMed2001

Isolation and spectroscopic characterization of a recombinant bell pepper hydroperoxide lyase.

Fatty acid hydroperoxide (HPO) lyase is a component of the oxylipin pathway and holds a central role in elicited plant defense. HPO lyase from bell pepper has been identified as a heme protein which shares 40% homology with allene oxide synthase, a cytochrome P450 (CYP74A). HPO lyase of immature bell pepper fruits was expressed in Escherichia coli and the enzyme was purified and characterized by spectroscopic techniques. The electronic structure and ligand coordination properties of the heme were investigated by using a series of exogenous ligands. The various complexes were characterized by using UV-visible absorption and electron paramagnetic resonance spectroscopy. The spectroscopic data demonstrated that the isolated recombinant HPO lyase has a pentacoordinate, high-spin heme with thiolate ligation. Addition of the neutral ligand imidazole or the anionic ligand cyanide results in the formation of hexacoordinate adducts that retain thiolate ligation. The striking similarities between both the ferric and ferrous HPO lyase-NO complexes with the analogous P450 complexes, suggest that the active sites of HPO lyase and P450 share common structural features.

Aldehyde-Lyases↗

PubMed2000

Antioxidative activity in the pericarp and seed of Japanese pepper (Xanthoxylum piperitum DC).

The antioxidative activity of the dried pericarp and seed of Japanese pepper was studied. The ethyl acetate extract from the pericarp and the methanol extract from the seed showed strong antioxidative activity against linoleic acid by the ferric thiocyanate and thiobarbituric acid (TBA) methods. Japanese pepper contained 3.9 and 2.9 mg/100 g of dry weight (dw) of tocopherols in the pericarp and seed, respectively, alpha-Toc in the former constituting 82% of total tocopherol and gamma-Toc in the latter constituting 96%. Arbutin and magnoflorine were isolated as antioxidants and their chemical structures determined by instrumental analyses. The contents of arbutin evaluated as the trifluoroacetate derivative by GC-MS were 35 and 3.0 mg/100 g of dw in the pericarp and seed, respectively. Magnoflorine was present only in the seed, and not in the pericarp. Both arbutin and magnoflorine exhibited antioxidative activity against linoleic acid and radical-scavenging activity against the DPPH radical.

Antioxidants↗

PubMed2001

Formation by mechanical stimulus of the flavor compounds in young leaves of Japanese pepper (Xanthoxylum piperitum DC.).

The volatile compounds formed in slapped and crushed young leaves of Japanese pepper (Xanthoxylum piperitum DC.) were compared with those of intact leaves by using a dynamic headspace gas analysis combined with GC-MS in an on-line system, together with the results of a sensory evaluation. The results indicated that the factors influencing the aroma note were mainly the constituent oxygenated monoterpenes and C(6) compounds and the quantity of terpene hydrocarbons. To clarify the formation mechanism for the aroma, the activities of lipoxygenase and beta-D-glucosidase were investigated. The results revealed that the hydrolysis of glycosides and the degradation of unsaturated fatty acids both played an important role in the formation of the major aroma compounds in young leaves of Japanese pepper.

Humans↗

PubMed2001

Isolation of some glycosides as aroma precursors in young leaves of Japanese pepper (Xanthoxylum piperitum DC.).

To clarify the formation mechanism for the major alcoholic aroma compounds in young leaves of Japanese pepper, the glycosides were isolated as aroma precursors. The presence of glycosides of the main alcoholic aroma constituents was indirectly determined by enzymatic hydrolysis and trifluoroacetylation (TFA) of the glycoside-containing fraction. After Amberlite XAD-2 column chromatography, ODS flash chromatography, and high-performance liquid chromatography (HPLC), two new compounds, namely, (3S,6S)-cis-linalool-3,7-oxide beta-D-glucopyranoside and 2-methylpropanyl 6-O-beta-D-apiofuranosyl-beta-D-glucopyranoside, were isolated. In addition, (3S,6R)-cis-linalool-3,6-oxide beta-D-glucopyranoside, which absolute configuration was the first determined, and six known glycosides, citronellyl beta-D-glucopyranoside, linalyl 6-O-beta-D-apiofuranosyl-beta-D-glucopyranoside, (Z)-3-hexenyl beta-D-glucopyranoside, benzyl 6-O-beta-D-apiofuranosyl-beta-D-glucopyranoside, dendranthemoside A, and 3,6-dihydroxy-5,6-dihydro-beta-ionol 9-beta-D-glucopyranoside, were isolated. All of these glycosides were isolated for the first time from the leaves of Japanese pepper. Their structures were established on the basis of spectral data and chemical evidence. The ratios of stereoisomers of the aglycon moieties of citronellyl beta-D-glucopyranoside and linalyl 6-O-beta-D-apiofuranosyl-beta-D-glucopyranoside were investigated by a chiral GC analysis and compared with those of free citronellol and linalool in the aroma concentrate.

Capsicum↗

PubMed2001

Structure-reactivity relationship in the oxidation of carotenoid pigments of the pepper (Capsicum annuum L.).

The relationship between the degradation rate and structure of each pigment of the pepper carotenoid profile was studied in mixtures of dehydrated fruit with lipid substrates of differing degrees of unsaturation and in different proportions (20 and 40%). The differences in structural nature of the carotenoids present in the pepper fruit produce a variable rate of oxidation, resulting in nonuniform degradation. The yellow xanthophylls and beta-carotene have the highest rates of oxidation, with the ketocarotenoids and violaxanthin degrading at lower rates. Autoxidation is greater or lesser depending on the functional groups, which stabilize the radical intermediaries of the reaction. The behavior of capsanthin and capsorubin is that expected of carotenoids having structures that include keto groups: a markedly greater stability to autoxidation processes. This increases their antioxidant capacity, adding to their beneficial impact by reducing the proliferation of radical processes, which are detrimental to health.

Capsicum↗