Purification, characterization and enzymatic synthesis of theaflavins of polyphenol oxidase isozymes from tea leaf (Camellia sinensis)

Teng, Jie; Gong, Zhihua; Deng, Yanli; Chen, Ling; Qin, Li; Shao, Yuanyuan; Li, Gang; Xiao, Wenjun

doi:10.1016/j.lwt.2017.05.065

Cited by 63 publications

(56 citation statements)

References 33 publications

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“…Extraction of enzyme extracts was based on our previous method with slight modifications ( Teng et al, 2017 ). 1.25-g tea leaves were ground with 0.75-g polyvinylpolypyrrolidone (PVPP) in a mortar on ice.…”

Section: Methodsmentioning

confidence: 99%

Biochemical Components Associated With Microbial Community Shift During the Pile-Fermentation of Primary Dark Tea

Qin

Chai

et al. 2018

Front. Microbiol.

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Primary dark tea is used as raw material for compressed dark tea, such as Fu brick tea, Hei brick tea, Hua brick tea, and Qianliang tea. Pile-fermentation is the key process for the formation of the characteristic properties of primary dark tea, during which the microorganism plays an important role. In this study, the changes of major chemical compounds, enzyme activities, microbial diversity, and their correlations were explored during the pile-fermentation process. Our chemical and enzymatic analysis showed that the contents of the major compounds were decreased, while the activities of polyphenol oxidase, cellulase, and pectinase were increased during this process, except peroxidase activity that could not be generated from microbial communities in primary dark tea. The genera Cyberlindnera, Aspergillus, Uwebraunia, and Unclassified Pleosporales of fungus and Klebsiella, Lactobacillus of bacteria were predominant in the early stage of the process, but only Cyberlindnera and Klebsiella were still dominated in the late stage and maintained a relatively constant until the end of the process. The amino acid was identified as the important abiotic factor in shaping the microbial community structure of primary dark tea ecosystem. Network analysis revealed that the microbial taxa were grouped into five modules and seven keystone taxa were identified. Most of the dominant genera were mainly distributed into module III, which indicated that this module was important for the pile-fermentation process of primary dark tea. In addition, bidirectional orthogonal partial least squares (O2PLS) analysis revealed that the fungi made more contributions to the formation of the characteristic properties of primary dark tea than bacteria during the pile-fermentation process. Furthermore, 10 microbial genera including Cyberlindnera, Aspergillus, Eurotium, Uwebraunia, Debaryomyces, Lophiostoma, Peltaster, Klebsiella, Aurantimonas, and Methylobacterium were identified as core functional genera for the pile-fermentation of primary dark tea. This study provides useful information for improving our understanding on the formation mechanism of the characteristic properties of primary dark tea during the pile-fermentation process.

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Section: Methodsmentioning

confidence: 99%

Biochemical Components Associated With Microbial Community Shift During the Pile-Fermentation of Primary Dark Tea

Qin

Chai

et al. 2018

Front. Microbiol.

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“…The optimum pH of cape gooseberry PPO was found to be 5.5 for catechol and 5.0 for chlorogenic acid (Bravo et al, 2016). Additionally, the optimum pH of the PPO enzyme from buriti pulp and tea leaf were determined to be pH 7.0 and 8.0 (Carvalho et al, 2017;Teng et al, 2017). Obviously, the optimal pH for PPO activity varies from approximately 5.0 to 8.0 depending on plant sources, the extraction methods, substrates and enzyme localization.…”

Section: Ph Optimum Temperature Optimum and Thermostabilitymentioning

confidence: 96%

“…Many common and effective inhibitors of PPO (Holzwarth, Wittig, Carle, & Kammerer, 2013) such as ascorbic acid, citric acid, sodium sulfite, phytic acid, sodium chloride and other compounds that have thiol groups (L-cysteine, glutathione) have been reported. More recently, the characterization of PPO has been studied in various plants food such as mung bean sprouts (Sikora & Świeca, 2018), bayberry juice (Cao, Cai, Wang, & Zheng, 2017), acai fruit (Jesus, Leite, & Cristianini, 2018), blueberry (Siddiq & Dolan, 2017), tea leaf (Teng et al, 2017), soursop nectar (Anaya-Esparza et al, 2017), cape gooseberry (Bravo & Osorio, 2016). However, few detailed studies have reported on the purification and characterization of water yam PPO.…”

Section: Introductionmentioning

confidence: 99%

Purification and characterization of polyphenol oxidase (PPO) from water yam (Dioscorea alata)

Peng

et al. 2019

CyTA - Journal of Food

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Polyphenol oxidase (PPO) is responsible for the enzymatic browning of fruits and vegetables, and thus has negative effects on the quality of food. In this study, extraction and purification of PPO from water yam was performed successfully, and the characteristics of PPO and inhibitors of enzyme activity were analysed. The results showed that PPO activity improved by 4.58-fold through the DEAE-Sepharose and Superdex G-75 purification technique, which reached electrophoretic purity. The enzyme molecular weight was estimated to be 32 kDa. The optimum pH and temperature values of PPO were 6.0 and 35°C, respectively, and it was not thermostable at higher temperatures. PPO had the highest substrate affinity towards catechol (Km = 27.82 mmol/L and Vmax = 1.464 △A/ min). The most effective inhibitor was L-cysteine followed by ascorbic acid and sodium sulfite. Purificación y caracterización de la polifenol oxidasa (PPO) obtenida del ñame de agua (Dioscorea alata) RESUMEN La polifenol oxidasa (PPO) es responsable del pardeamiento enzimático de frutas y verduras; por lo tanto, tiene efectos negativos en la calidad de los alimentos. En el presente estudio se extrajo y se purificó con éxito la PPO proveniente del ñame de agua, y se analizaron sus características y los inhibidores de la actividad enzimática. En este sentido, los resultados indicaron que la actividad de la PPO mejoró 4.58 veces al emplear la técnica de purificación DEAE-Sepharose y Superdex G-75, que alcanzó la pureza electroforética. El peso molecular de la enzima se estimó en 32 kDa. Los valores óptimos de pH y temperatura de la PPO fueron 6.0 y 35°C, respectivamente, constatándose que no era termoestable a temperaturas más altas. La PPO mostró mayor afinidad de sustrato hacia el catecol (Km = 27.82 mmol/L y Vmax = 1.464 △A/min). Su inhibidor más efectivo fue la L-cisteína, seguida por el ácido ascórbico y el sulfito de sodio.

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“…PPOs play multiple roles in C. sinensis , such as the elicitation of defenses against E. obliqua and the oxidation of flavanols to theaflavins and thearubigins during tea processing [4,38,39]. However, which members of the CsPPO gene family are involved in the defense response of tea plants have not yet been identified.…”

Section: Discussionmentioning

confidence: 99%

Two New Polyphenol Oxidase Genes of Tea Plant (Camellia sinensis) Respond Differentially to the Regurgitant of Tea Geometrid, Ectropis obliqua

Huang

Jin

Zhang

et al. 2018

IJMS

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Polyphenol oxidases (PPOs) have been reported to play an important role in protecting plants from attacks by herbivores. Though PPO genes in other plants have been extensively studied, research on PPO genes in the tea plant (Camellia sinensis) is lacking. In particular, which members of the PPO gene family elicit the defense response of the tea plant are as yet unknown. Here, two new PPO genes, CsPPO1 and CsPPO2, both of which had high identity with PPOs from other plants, were obtained from tea leaves. The full length of CsPPO1 contained an open reading frame (ORF) of 1740 bp that encoded a protein of 579 amino acids, while CsPPO2 contained an ORF of 1788 bp that encoded a protein of 595 amino acids. The deduced CsPPO1 and CsPPO2 proteins had calculated molecular masses of 64.6 and 65.9 kDa; the isoelectric points were 6.94 and 6.48, respectively. The expression products of recombinant CsPPO1 and CsPPO2 in Escherichia coli were about 91 and 92 kDa, respectively, but the recombinant proteins existed in the form of an inclusion body. Whereas CsPPO1 is highly expressed in stems, CsPPO2 is highly expressed in roots. Further results showed that the expression of CsPPO1 and CsPPO2 was wound- and Ectropis obliqua-induced, and that regurgitant, unlike treatment with wounding plus deionized water, significantly upregulated the transcriptional expression of CsPPO2 but not of CsPPO1. The difference between regurgitant and wounding indicates that CsPPO2 may play a more meaningful defensive role against E. obliqua than CsPPO1. Meanwhile, we found the active component(s) of the regurgitant elicited the expression of CsPPO may contain small molecules (under 3-kDa molecular weight). These conclusions advance the understanding of the biological function of two new PPO genes and show that one of these, CsPPO2, may be a promising gene for engineering tea plants that are resistant to E. obliqua.

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Purification, characterization and enzymatic synthesis of theaflavins of polyphenol oxidase isozymes from tea leaf (Camellia sinensis)

Cited by 63 publications

References 33 publications

Biochemical Components Associated With Microbial Community Shift During the Pile-Fermentation of Primary Dark Tea

Biochemical Components Associated With Microbial Community Shift During the Pile-Fermentation of Primary Dark Tea

Purification and characterization of polyphenol oxidase (PPO) from water yam (Dioscorea alata)

Two New Polyphenol Oxidase Genes of Tea Plant (Camellia sinensis) Respond Differentially to the Regurgitant of Tea Geometrid, Ectropis obliqua

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