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排序方式: 共有211条查询结果,搜索用时 31 毫秒
71.
Tea leaf is rich in pyrogallol-type catechins, and their oxidation is important in the generation of black tea polyphenols. In the present study, the enzymatic oxidation of three pyrogallol-type catechins, (+)- and (−)-gallocatechins and (−)-epigallocatechin, was compared. The reactions yielded unstable quinone products, which were trapped as condensation products with o-phenylenediamine. The oxidation of (+)-gallocatechin proceeded very slowly compared to the reaction of (−)-epigallocatechin, and yielded a proepitheaflagallin-type dimer as the major product, though oxidation of (−)-epigallocatechin gave predominantly dehydrotheasinensin C. The cis-configuration of the C-3 hydroxyl group and the B-ring of (−)-epigallocatechin was apparently crucial for rapid and selective production of dehydrotheasinensin C. Oxidation of (−)-gallocatechin proceeded in a manner similar to that of (+)-gallocatechin, and produced an enantiomer of the (+)-gallocatechin product. The results suggest that enzymes catalyze oxidation of the pyrogallol B-ring to the o-quinone, with subsequent non-enzymatic coupling reactions proceed under highly steric control. 相似文献
72.
73.
In this study, polyphenol oxidase (PPO) was extracted from Prunus domestica and partially purified by ammonium sulfate precipitation, hydrophobic interaction chromatography, and ion exchange chromatography. The final purification step revealed a 32.81-fold purification, and the molecular mass was estimated to be 65 kDa by SDS-PAGE. The purified PPO showed enzymatic activity mainly toward five substrates, namely catechol, catechin, 4-methyl catechol, chlorogenic acid, and L-3,4-dihydroxyphenylalanine, whereas it showed no activity toward caffeic acid, ferulic acid, p-coumaric acid, p-cresol, and l-tyrosine. The optimum pH and temperature values were 6.0 and 25 °C, respectively. The enzyme showed high stability in the pH range of 5.0–7.0 and in the temperature range of 25–65 °C. The most effective inhibitors of this enzyme were found to be ascorbic acid and l-cysteine. The thermal inactivation followed a first-order kinetic model, with activation energy of Ea 150.46 ± 1.29 kJ/mol. PPO extracted from plum showed stability at high pressure, with enzyme activation at 500 MPa. 相似文献
74.
Polyphenol oxidases (PPOs) are copper-containing industrially important enzymes that catalyze the synthesis of many commercially important products by using polyphenols as substrate. Camellia sinensis polyphenol oxidase (CsPPO) is interesting because it oxidizes epicatechins to yield theaflavins and thearubigins. The present study aimed to optimize the expression of CsPPO in Escherichia coli. Because CsPPO had a large number of E. coli rare codons, it yielded a poor quantity of protein in E. coli Rosetta™ 2 cells, which have additional tRNAs for E. coli rare codons. Thus, synthetically constructed codon-optimized CsPPO was cloned into pET-47b(+) vector and expressed in a bacterial host. Ectopic expression led to the formation of inclusion bodies. However, extensive standardization of buffers and methods of refolding such as dialysis, on-column refolding, and rapid dilution yielded active PPO from solubilized inclusion bodies with copper content of 0.880 ± 0.095 atom/molecule of protein.Experimental data produced maximum PPO activity in a rapid dilution buffer containing 0.5 M L-arginine. Refolded CsPPO had an optimum pH of 5.0 and Km values of 3.10, 0.479, and 0.314 mM, and a Vmax of 163.9, 82.64, and 142.8 U/mg of protein for catechol, catechin, and epicatechin, respectively. 相似文献
75.
The structure of tyrosinase (Tyr) is reviewed from a double point of view. On the one hand, by comparison of all Tyr found throughout nature, from prokaryotic organisms to mammals and on the other, by comparison with the tyrosinase related proteins (Tyrps) that appeared late in evolution, and are only found in higher animals. Their structures are reviewed as a whole rather than focused on the histidine (His)‐bound metal active site, which is the part of the molecule common to all these proteins. The availability of crystallographic data of hemocyanins and recently of sweet potato catechol oxidase has improved the model of the three‐dimensional structure of the Tyr family. Accordingly, Tyr has a higher structural disorder than hemocyanins, particularly at the CuA site. The active site seems to be characterized by the formation of a hydrophobic pocket with a number of conserved aromatic residues sited close to the well‐known His. Other regions specific of the mammalian enzymes, such as the cytosolic C‐terminal tail, the cysteine clusters, and the N‐glycosylation sequons, are also discussed. The complete understanding of the Tyr copper‐binding domain and the characterization of the residues determinant of the relative substrate affinities of the Tyrps will improve the design of targeted mutagenesis experiments to understand the different catalytic capabilities of Tyr and Tyrps. This may assist future aims, from the design of more efficient bacterial Tyr for biotechnological applications to the design of inhibitors of undesirable fruit browning in vegetables or of color skin modulators in animals. 相似文献
76.
Daniel Lpez‐serrano Antonio SAnchez‐Amat Francisco Solano 《Pigment cell & melanoma research》2002,15(2):104-111
The sequence of the tyrosinase gene cloned from Marinomonas mediterranea is reported. It is the second tyrosinase cloned from a Gram negative bacterium. Its size is higher than that of Gram positive tyrosinases from Streptomyces, and more similar to the eukaryotic enzymes. Its sequence shares the features of copper‐binding sites found in all tyrosinases. Based in the comparison of tyrosinases from all types of organisms, an extension of the characteristic signatures existing at Prosite is proposed. This tyrosinase shares with some plant and amphibian tyrosinases a strong specific activation by submicellar concentrations of SDS. Intrinsic fluorescence and kinetic properties indicate that the activation is caused by an SDS‐dependent conformational change that facilitates the substrate accessibility to the dicopper active site. 相似文献
77.
不同品种南瓜POD及PPO同工酶的比较研究 总被引:17,自引:0,他引:17
采用垂直平板聚丙烯酰胺凝胶电泳技术,对美洲南瓜(Cucurbita pepo L.)和中国南瓜(C.moschata Duch.)共10个不同栽培品种的过氧化物酶(POD)及多酚氧化酶(PPO)同工酶进行了分析研究,并采用聚类分析法对其亲缘关系进行了比较和分类。结果表明,10个不同品种的南瓜根POD共分离出10条酶带,其中el、e4、e5、e6和e9为2个栽培种所共有;e4、e5、e6分离清晰,可作为2个栽培种的特征酶带;PPO同工酶分离出6条酶带,其中e′1和e′4分离清晰,为2个栽培种所共有。 相似文献
78.
Effects of putrescine and ethephon on some oxidative stress enzyme activities and proline content in salt stressed spinach leaves 总被引:1,自引:0,他引:1
The effects of putrescine and ethephon on peroxidase (POD; EC 1.11.1.7), polyphenol oxidase (PPO; EC 1.14.18.1), catalase (CAT; EC 1.11.1.6) activities and proline content in spinach leaves under saline stress were investigated. In control conditions, putrescine increased PPO and CAT activities and proline content, but decreased POD activity. Ethephon increased these three enzyme activities but did not affect proline content. In saline conditions, putrescine increased POD and CAT activities and proline content, while it decreased PPO activity. Ethephon increased both PPO and CAT activities and proline content, but decreased POD activity. Putrescine and ethephon have opposite effects on the enzyme activities and proline accumulation because they acts as antagonists. 相似文献
79.
为探讨嘉宝果(Myrciaria cauliflora)叶片多酚的分离纯化方法,对4种树脂(NKA-2、NKA-9、HPD-826和HPD-400A)进行了筛选,并分析了其多酚的抗氧化、体外降糖活性和组成成分。结果表明,NKA-9树脂适于嘉宝果叶片多酚纯化,最佳工艺条件为:上样液质量浓度2.00 mg/mL、洗脱液乙醇体积分数70%、上样流速1.0 mL/min、上样量204 mL、洗脱流速0.9 mL/min、洗脱量70 mL。嘉宝果叶多酚纯度可达69.86%。嘉宝果叶片纯化后的多酚抗氧化及α-葡萄糖苷酶抑制活性高于纯化前,但α-淀粉酶抑制活性低于纯化前。HPLC结果表明,嘉宝果叶片中含有杨梅苷、芦丁、金丝桃苷和鞣花酸,其中鞣花酸含量最高[(16.15±0.49) mg/g]。因此,NKA-9树脂适合分离纯化嘉宝果叶片多酚,纯化后的多酚抗氧化及α-葡萄糖苷酶抑制活性增强。 相似文献
80.
Dong-Hyun Lee Sung-Sik Choi Bo-Bae Kim Se-Young Kim Bo-Sik Kang Sung-Joon Lee Hyun-Jin Park 《Process Biochemistry》2013,48(12):1964-1971
Polyphenolic composition of alcohol-free red wine concentrates (AFRWC) was determined by LC–MS/MS. The concentration of salicylic acid in non-flavonoid class and malvidin in flavonoid class was the highest among all the polyphenols determined in AFRWC. In the in vitro model using HepG2 cells, AFRWC was found to be more effective for the reduction of total cholesterol than lovastatin. For the in vivo model, animals were provided with AFRWC at ~750 mg of total polyphenols/kg body weight per day by oral administration. The amount of AFRWC was established by extrapolation to be equivalent to 375 ml/day wine consumption, which is ~2–3 glasses of wine per day for a 60 kg human. Despite a high cholesterol diet, a significant reduction in both total cholesterol and LDL-cholesterol was observed when supplemented with AFRWC, but the increase of HDL-cholesterol was not observed. The expression level of mRNA of some hepatic genes participating in cholesterol biosynthesis, cholesterol esterification was found to be influenced by AFRWC supplementation, whereas reverse cholesterol transport involved with HDL-cholesterol was seldom affected showing discrepancy in the expression of associated genes. 相似文献