Purification and partial characterization of a peroxidase from plant cell cultures of Cassia didymobotrya and biotransformation studies

Vitali, Alberto; Botta, Bruno; Monache, Giuliano Delle; Zappitelli, Sabrina; Ricciardi, P.; Melino, Sonia; Petruzzelli, Raffaele; Giardina, Bruno

doi:10.1042/bj3310513

Cited by 34 publications

(20 citation statements)

References 25 publications

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“…The presence of seven phenyl protons and twelve phenyl carbons was confirmed with a 1D-NMR spectrum, and a keto group (d = 190.7) was also detected in the 13 C-NMR spectrum. In addition, 1 was also analyzed by HMQC and HMBC experiments, and the obtained data for 1 were very similar to the reported data for the meso form of 3,3¢¢-di(7,4¢-dihydroxyflavanone-3-yl) which had been obtained only as the biotransformation product [8], [9]. Although the signals of H-2, H-2¢¢, H-3 and H-3¢¢ almost could not be observed due to the fixed conformation at 23 8C, these appeared as doublets (d H = 4.86, 2H, d, J = 12.0 Hz, H-2/H-2¢¢ and 3.55, 2H, d, J = 12.0 Hz, H-3/H-3¢¢) on NMR analysis at 80 8C in DMSO-d 6 in a similar manner as reported previously [9], suggesting that H-2, H-2¢¢, H-3 and H-3¢¢ were each located at axial sites.…”

supporting

confidence: 56%

“…Compound 2 gave similar spectra upon 1 H-NMR, 13 C-NMR and MS analyses as the other biotransformation product, the diastereomer of 1 [9]. Although this biotransformation product of 1 has been reported as the racemate, which showed no optical rotation [9], 2 showed significant dextrorotation (+ 83.28 in MeOH at 25 8C). These findings suggested that 2 might be a diastereomer of 1, but the absolute stereostructure of 2 could not be determined in the present study.…”

Section: Resultsmentioning

confidence: 74%

“…These findings suggested that 2 might be a diastereomer of 1, but the absolute stereostructure of 2 could not be determined in the present study. Compound 1 and the racemate of 2 had been obtained only as biotransformation products from chalcones with peroxidases of cultured plant cells [8], [9], [10]. …”

Section: Resultsmentioning

confidence: 99%

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Antimalarial Activity of Biflavonoids from Ochna integerrima

Ichino¹,

Kiyohara²,

Soonthornchareonnon³

et al. 2006

Planta med

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IntroductionMalaria is the major parasitic infection in many tropical and subtropical regions, leading to more than one million deaths (principally among African children) out of 400 million cases each year [1] and to major consequent impacts on economic productivity and livelihood [2]. The incidence of malaria is now increasing because of the appearance of multi-drug resistant Plasmodium falcipalum, therefore new and more effective antimalarial drugs are urgently required.Ochna integerrima Merr. (Ochnaceae) is a tree that is widely distributed in Thailand [3], and the bark of O. integerrima Merr. has been used for digestive disorders as a folk medicine in Thailand [4]. In phytochemical studies on O. integerrima Merr, many flavonoids have been isolated [5], [6], [7]. During a screening for antimalarial active sources among Thai plants, it was found that the 80 % EtOH extract from the outer bark of O. integerrima Merr.showed significant anti-malarial activity. The active ingredients in the outer bark were clarified in the present study. Materials and Methods ApparatusOptical rotations were measured on a JASCO polarimeter. 1 H-and 13 C-NMR spectra were determined on a Varian Mercury-300. Mass spectra (MS) were obtained on JEOL MXA-AM505HA and JMS-700 MStation spectrometers. Chromatographic separations were carried out by column chromatography on Wakogel C-200 (75 ± 150 mm, Wako; Osaka, Japan). Preparative reverse phase HPLC was carried out on PEGASIL (250 20 mm i. d., Senshu Co. Ltd.; Tokyo, Japan). Antimalarial Activity of Biflavonoids from Ochna integerrimaChikara AbstractDuring the screening of antimalarial substances, the 80 % EtOH extract from the outer bark of Ochna integerrima Merr. (Ochnaceae) was shown to have a good anti-malarial activity (IC 50 value: 6.5 mg/mL) whereas extracts from the inner barks of O. integerrima showed no antimalarial activity. Biflavanone (1), which had not been found previously from a natural plant source, was isolated as a potent antimalarial active ingredient (IC 50 value: 80 ng/mL) from the extract of the outer barks. The stereoisomer of 1 ( = compound 2) was also isolated from this plant; however, its activity was significantly lower than that of 1. Key wordsAntimalarial´Ochnaceae´Ochna integerrima Merr.´biflavanones Original Paper 611This document was downloaded for personal use only. Unauthorized distribution is strictly prohibited. Analytical methodsAnalytical HPLC experiments were performed on an Agilent 1100 series HPLC instrument (Agilent Technologies Japan, Ltd.; Tokyo, Japan) equipped with a column of PEGASIL (250 4.6 mm i. d., Senshu Co. Ltd.; Tokyo, Japan). The solvent system used was a linear gradient of acetonitrile from 30 % to 60 % during 30 minutes in 10 mM phosphoric acid. Flow rate was 1 mL/min. Injection volume was 10 mL for MeOH solution of all the samples. Concentrations of the samples were 10 mg/mL for extracts and 1 mg/ mL for the isolated compounds. Plant material Extraction and isolationThe outer barks of O. integerrima (485 g) were extracted with 80...

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supporting

confidence: 56%

Section: Resultsmentioning

confidence: 74%

See 1 more Smart Citation

Antimalarial Activity of Biflavonoids from Ochna integerrima

Ichino¹,

Kiyohara²,

Soonthornchareonnon³

et al. 2006

Planta med

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“…The reaction was started by the addition of the protein extract at room temperature. Enzymatic activity was calculated with an extinction coefficient of 26.6 mM 21 cm 21 at 470 nm (Vitali et al, 1998). Values are means 6 SE of three replicates.…”

Section: Enzyme Assaysmentioning

confidence: 99%

“…Enzyme activity was assayed according to a published procedure (Vitali et al, 1998) in solutions consisting of 2.8 mL of buffer (0.1 M potassium phosphate buffer, pH 7.0), 50 mL of substrate (18 mM), 50 mL of H 2 O 2 (30%, 1:100 diluted, optical density at 240 nm = 0.4), and 100 mL of crude FaPRX27 extract (1 mg mL 21 ). Reactions were monitored photometrically at the wavelength of maximum absorption of the respective substrates.…”

Section: Enzyme Assaysmentioning

confidence: 99%

Metabolic Interaction between Anthocyanin and Lignin Biosynthesis Is Associated with Peroxidase FaPRX27 in Strawberry Fruit

et al. 2013

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Plant phenolics have drawn increasing attention due to their potential nutritional benefits. Although the basic reactions of the phenolics biosynthetic pathways in plants have been intensively analyzed, the regulation of their accumulation and flux through the pathway is not that well established. The aim of this study was to use a strawberry (Fragaria × ananassa) microarray to investigate gene expression patterns associated with the accumulation of phenylpropanoids, flavonoids, and anthocyanins in strawberry fruit. An examination of the transcriptome, coupled with metabolite profiling data from different commercial varieties, was undertaken to identify genes whose expression correlated with altered phenolics composition. Seventeen comparative microarray analyses revealed 15 genes that were differentially (more than 200-fold) expressed in phenolics-rich versus phenolics-poor varieties. The results were validated by heterologous expression of the peroxidase FaPRX27 gene, which showed the highest altered expression level (more than 900-fold). The encoded protein was functionally characterized and is assumed to be involved in lignin formation during strawberry fruit ripening. Quantitative trait locus analysis indicated that the genomic region of FaPRX27 is associated with the fruit color trait. Down-regulation of the CHALCONE SYNTHASE gene and concomitant induction of FaPRX27 expression diverted the flux from anthocyanins to lignin. The results highlight the competition of the different phenolics pathways for their common precursors. The list of the 15 candidates provides new genes that are likely to impact polyphenol accumulation in strawberry fruit and could be used to develop molecular markers to select phenolics-rich germplasm.

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Anionic peroxidase production byArnebia euchromacallus

Farhadi¹,

Haghbeen²,

Marefatjo³

et al. 2011

Biotech and App Biochem

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Arnebia euchroma callus, obtained from the root cell culture of an Iranian native specimen, has gained a doubling time of 63 H after regular subculturing on Linsmaier-Skoog (LS) medium containing sugar (50 g/L), 2,4-dichlorophenoxyacetic acid (10(-6) M), and kinetin (10(-5) M) under darkness at 25°C. Despite the observed somaclonal variations, peroxidase production by the A. euchroma calli has been stable over 4 years under the aforementioned conditions. Isoelectric focusing experiments revealed that the partially purified A. euchroma peroxidases (AePoxs) are mainly anionic with pI values of about 5.5 and 6.6. AePox reaches its optimal activity at 55°C and pH 7.5. Results of the various kinetic studies suggest that AePox belongs to the type III plant peroxidases with no activity for the oxidation of 3-indoleacetic acid, but seems to play a role in the lignin biosynthesis and H(2) O(2) regulation during the proliferation of the A. euchroma cells on LS medium. Comparing the biochemical properties of AePox with horseradish peroxidase and in view of the ease of solid cell culture, the A. euchroma callus could be considered as a source of plant peroxidase for some biotechnological applications.

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Purification and partial characterization of a peroxidase from plant cell cultures of Cassia didymobotrya and biotransformation studies

Cited by 34 publications

References 25 publications

Antimalarial Activity of Biflavonoids from Ochna integerrima

Antimalarial Activity of Biflavonoids from Ochna integerrima

Metabolic Interaction between Anthocyanin and Lignin Biosynthesis Is Associated with Peroxidase FaPRX27 in Strawberry Fruit

Anionic peroxidase production byArnebia euchromacallus

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