A reliable technique to identify superior quality clones from tea germplasm

Variations in the substrate level and enzyme activity of prominent south Indian tea germplasm were studied. The content of polyphenols, catechins (substrates) and polyphenol oxidase (PPO) showed variation, which influenced the final black tea quality. The enzyme PPO occurs in tea shoots and catalyzes the reactions between catechins to form theaflavins in the presence of oxygen. The catechins mainly epicatechin (EC), epigallo catechin gallate (EGCG), epicatechin gallate (ECG) and epigallo catechin (EGC) get mixed with PPO during the oxidation process to form quality constituents like theaflavins (TF), thearubigins (TR) and high polymerized substances (HPS). Theaflavins and their fractions such as simple theaflavin, theaflavin monogallate, (TFMG), theaflavin digallate (TFDG) in black tea are the essential quality constituents that are responsible for the liquor characteristics where as TR and high polymerized substances impart colour to the liquor. As oxidation of macerated leaves proceed through different stages of tea manufacture, a decline in PPO activity, polyphenol and catechin contents were observed. Data revealed that the oxidation reaction was faster during the initial stages of oxidation. During the period, oxygen consumption was higher and declined thereafter. Ratio between the enzyme (PPO) and its substrate (catechins) were used to characterize the quality potential of tea clones. An attempt has also been made to categorize prominent tea clones as high, moderate and average quality clones based on their enzyme substrate ratio. Theaflavin content (oxidation product) of different tea clones suggests that the ratio between PPO and catechins forms an important criterion which determines black tea quality. Results obtained were compared with standard clones of known high quality (CR-6017) and moderate quality (SA-6). The study reveals that the enzyme substrate ratio can be used to identify superior quality clones from the existing tea germplasm.     

茶制品的发展现状及趋势

茶叶的深加工与综合利用是未来茶产业可持续发展的关键所在,茶制品的发展给整个茶行业带来了巨大的经济效益。目前茶制品主要形式为以茶饮料(茶汤)、调味茶饮料、复(混)茶饮料等茶品形式的茶饮料,以茶餐、茶味坚果、茶味糖果、茶味糕点等风味化的茶食品,以茶功能性成分如茶多酚、咖啡碱、茶氨酸等为主体的保健品,以茶味牙膏、茶枕、茶袜子等形式的日用品。随着现代食品加工技术的发展,茶资源的利用将更加充分、深入,茶制品将有以下三种发展趋势:一是可利用的茶树资源的丰富化;二是以茶叶功能性成分为主的保健品的开发;三是产品的多元化。     

六堡茶渥堆过程中产多酚氧化酶菌株的筛选及多酚氧化酶的分离纯化

本研究采用传统分离法从渥堆六堡茶中共分离出42株菌,然后利用平板初筛和液体发酵进行复筛,从42株微生物中筛选出10株产PPO的微生物并测定其酶活.用盐析及透析法对产酶活性最高的菌株所产的多酚氧化酶进行纯化,纯化后的酶液用SDS-PAGE及Native-PAGE电泳方法确定PPO的分子量范围和PPO种类.结果表明,渥堆六...     

六堡茶发酵前后多酚氧化酶的分离及酶学性质

为探索六堡茶发酵过程中多酚氧化酶(polyphenol oxidase,PPO)的来源及其酶学活性,采用不同pH值缓冲液、保护剂和液料比对发酵前后六堡茶PPO进行提取,通过(NH4)2SO4沉淀、透析和Sephadex?G-100凝胶过滤层析纯化PPO.结果表明:从六堡茶中分离纯化得到2个PPO同工酶,分子质量分别为5...     

The effect of withering on fermentation of tea leaf and development of liquor characters of black teas

Depression in polyphenol oxidase (PPO) activity during the withering of tea leaf (Camellia sinensis) in black tea manufacture affected the oxidative-condensation of tea flavanols in forming theaflavins (TF) and thearubigins (TR), which are associated with brightness, briskness and ‘body’ of tea liquors. In contrast to conventionally manufactured CTC teas, unwithered fresh leaf CTC produces a higher proportion of TF and lower TR, resulting in bright, brisk and thin liquors. Hardness of wither was accompanied by a further depression in PPO activity, and formation of TF also declined with a concomitant loss in brightness and briskness. The TR content increased up to a certain degree of wither and thereby improved the ‘body’ of the liquor. Very hard wither, accompanied by a large reduction in moisture content, restricts PPO activity. It appears that in the fresh leaf, high moisture levels together with dissolved oxygen accelerate the enzymic oxidation and produce large amounts of TF. Senescence of the leaf seems to facilitate the production of TR.     

Quality evaluation of black CTC teas based upon seasonal variations

Effects of seasonal variations on the colour and chemical constituents of Indian black tea were investigated. The experimentation involved collection of different black tea samples that were cultivated at the same tea garden but harvested in three different seasons. Measurement of moisture content was done along with theaflavins (TFs), thearubigins (TRs) and colour (CIE La*b* values) of the brewed tea liquor using spectrophotometric approach. The correlation analysis between chemical constituents, that is, TFs and TRs, moisture content, colour and quality grading was done by using principal component analysis (PCA). The results suggest that quality of tea is dependent upon seasonal variations in terms of chemical changes such as TFs, TRs and moisture content, which also correlate well with colour. The efficacy of the colour of brewed tea liquor in explaining the changes in tea quality due to seasonal variations can be beneficial for scientific fixation of prices at tea auction centres.     

Prokaryotic expression and purification of Camellia sinensis polyphenol oxidase

BACKGROUND: Polyphenol oxidase (PPO) causes the postharvest loss of fruits and vegetables but is also a key factor in the quality development of tea. However, there are no reports on engineered active plant PPO purified from prokaryotic cells. RESULTS: In this study the ppo gene of about 1800 bp from Camellia sinensis cv. Yihongzao was successfully cloned and expressed in Escherichia coli. The PPOs purified from both the soluble fraction and the inclusion bodies showed activity. In addition, 1.0 × 10^?7 mol L^?1 Cu²⁺ and acidic conditions were found to be favourable for the engineered PPO catalysis of catechol oxidation. CONCLUSION: This paper represents the first report on C. sinensis ppo expression in E. coli and engineered active PPO purification. The results of the study provide a basis for the large‐scale preparation and application of PPO. Copyright © 2010 Society of Chemical Industry     

茶叶多酚氧化酶三相分离纯化及酶学性质研究

为研究一种快速高效的茶叶多酚氧化酶(polyphenol oxidase,PPO)制备方法,本试验采用三相分离法(three phase partitioning,TPP)获取茶叶PPO,并研究其酶学特性.结果表明,通过两次三相分离纯化后可获得酶比活力210.46 U/mg、纯化15.76倍、回收率8.04％的茶叶PP...     

The effect of process parameters on seasonal development of flavour in black tea

Analysis by g.l.c. of ether–extractable steam volatile constituents of black tea from a high elevation tea estate in the Western Region (Dimbulla District) of Sri Lanka during seasonal flavour development, indicated the presence of trans-2-hexenal, n-butyraldehyde, cis-3-hexenol, 1-octen-3-ol linalool, methyl salicylate and geraniol as the main flavour constituents. Of these, trans-2-hexenal and linalool showed appreciable seasonal changes in black tea. Leaf quality and manufacturing practices, particularly withering and fermentation, influenced the production of these compounds during black tea manufacture. It is suggested that the relative concentration of these compounds may be partly responsible for the typical flavour quality of Sri Lanka black teas and these may be used as indices of flavour development.     

微生物资源在茶产业中的应用现状及展望

随着生态农业发展和现代生物技术的不断进步,茶产业进入转型发展期,走生态、绿色、环保的环境友好型和资源节约型发展模式,是茶产业可持续发展的重要内容。微生物作为一种新型环保生物资源,由于其在长期的进化过程所形成物种多样性和功能多样性,加上其结构简单便于研究应用的特性,许多有益农业微生物已被开发成微生物肥料、微生物农药、微生物保鲜剂、微生物降污剂等资源,在茶叶无公害种植生产,茶叶加工和茶叶副产物资源的综合利用等方面具有广泛的应用。本文就微生物资源在茶产业中肥培管理、病虫害防治、农药残留绿色防控、茶叶加工、储藏保鲜剂以及茶综合利用方面的国内外研究进展进行概述,同时探讨微生物资源在茶产业应用中存在的问题。     

Ruminal micro‐organisms do not adapt to increase utilization of poly‐phenol oxidase protected red clover protein and glycerol‐based lipid

BACKGROUND: The enzyme polyphenol oxidase (PPO) reduces the extent of proteolysis and lipolysis within red clover fed to ruminants with subsequent increases in the efficiency of N utilization and the level of beneficial polyunsaturated fatty acids in their products (meat and milk). It has also been reported that red clover feeding alters the rumen microbial population compared to grass feeding. This study investigated whether the observed shifts in the microbial population of the rumen when ruminants are fed red clover silage (RC) as opposed to grass silage (G) represented an adaptation by the micro‐organisms to increase the utilization of PPO‐protected protein and glycerol‐based lipid. RESULTS: The experiment consisted of two periods where ruminally fistulated dairy cows were offered either RC or G for 2 weeks, followed by collection of rumen fluid, which was then used in in vitro incubations to investigate lipolysis and proteolysis over time in plant material derived from red clover plants with either wild type PPO expression (PPO+) or PPO expression reduced to undetectable levels by gene silencing (PPO?). Proteolysis and lipolysis (P < 0.05) were lower after 24 h of incubation in the PPO+ treatment than the PPO? treatment irrespective of rumen fluid. Biohydrogenation of C18 polyunsaturated fatty acids was also lower on the PPO+ treatment than the PPO? treatment, with no effect of rumen fluid. CONCLUSION: These results suggest that microbial changes to red clover feeding did not result in an increased ability of the micro‐organisms in the present study to utilize either PPO‐protected protein or glycerol‐based lipid. Copyright © 2008 Society of Chemical Industry     

Immunogold labelling to localize polyphenol oxidase (PPO) during wilting of red clover leaf tissue and the effect of removing cellular matrices on PPO protection of glycerol‐based lipid in the rumen

BACKGROUND: The enzyme polyphenol oxidase (PPO) reduces the extent of proteolysis and lipolysis within red clover fed to ruminants. PPO catalyses the conversion of phenols to quinones, which can react with nucleophilic cellular constituents (e.g. proteins) forming protein–phenol complexes that may reduce protein solubility, bioavailability to rumen microbes and deactivate plant enzymes. In this study, we localized PPO in red clover leaf tissue by immunogold labelling and investigated whether red clover lipid was protected in the absence of PPO‐induced protein–phenol complexes and plant enzymes (lipases). RESULTS: PPO protein was detected to a greater extent (P < 0.001) within the chloroplasts of mesophyll cells in stressed (cut/crushed and wilted for 1 h) than freshly cut leaves for both palisade (61.6 and 25.6 Au label per chloroplast, respectively) and spongy mesophyll cells (94.5 and 40.6 Au label per chloroplast, respectively). Hydrolysis of lipid and C18 polyunsaturated fatty acid biohydrogenation during in vitro batch culture was lower (P < 0.05) for wild‐type red clover than for red clover with PPO expression reduced to undetectable levels but only when cellular matrices containing protein–phenol complexes were present. CONCLUSION: Damaging of the leaves resulted in over a doubling of PPO detected within mesophyll cells, potentially as a consequence of conversion of the enzyme from latent to active form. PPO reduction of microbial lipolysis was apparent in macerated red clover tissue but not in the absence of the proteinaceous cellular matrix, suggesting that the PPO mechanism for reducing lipolysis may be primarily through the entrapment of lipid within protein–phenol complexes. Copyright © 2009 Society of Chemical Industry     

Enhancement of Fermentation Process in Pu-Erh Tea by Tea-Leaf Extract

ABSTRACT:  Pu-erh tea is known as a fermented tea and longer storage enhances its flavor and taste. Recently,  Aspergillu s,  Blastobotrys , and  Streptomyces  are found to play important roles in nutritional enhancement of Pu-erh tea by fermentation. Since water and temperature affect the microbial growth, we therefore explored the factors that might enhance the Pu-erh tea fermentation. The results showed that the addition of fresh tea-leaf extract (TLE) enhanced the withered tea fermentation (at 37 °C, 80 to 85% RH) as compared with the water only. Contents of statin, GABA, gallic acid, DPPH scavenging and polyphenol oxidase (PPO) activities were increased, whereas polyphenols and caffeine were decreased over 6 mo. TLE dose-dependently enhanced some of the qualities (that is, statin, PPO) of Pu-erh tea significantly as compared with the water only. The effect was related to the increase population of  A. niger  and  A. carbonarius  at 6 mo (from 7.6 ± 1.2 × 10¹ and 3.2 ± 1.3 × 10¹ to 3.1 ± 1.2 × 10⁶ and 2.4 ± 1.1 × 10⁵ colony forming units [CFU]/g, respectively). After drying process (90 °C, 30 min), the total microbial count from these samples returned to background level (3 ± 0.5 × 10² CFU/g). None of ochratoxin and fumonisin, toxins from  Aspergillus , was detected in the final products. The flavor and taste were also enhanced by treatment with TLE. The inoculation with  S. cinereus  Y11 with 2% TLE further enhanced these functional contents (about 2-fold increase of statin level) in the experimental Pu-erh tea. Therefore, this result may add a new process for Pu-erh tea manufacture.     

Deodorization of Garlic Breath by Foods,and the Role of Polyphenol Oxidase and Phenolic Compounds

Garlic causes a strong garlic breath that may persist for almost a day. Therefore, it is important to study deodorization techniques for garlic breath. The volatiles responsible for garlic breath include diallyl disulfide, allyl mercaptan, allyl methyl disulfide, and allyl methyl sulfide. After eating garlic, water (control), raw, juiced or heated apple, raw or heated lettuce, raw or juiced mint leaves, or green tea were consumed immediately. The levels of the garlic volatiles on the breath were analyzed from 1 to 60 min by selected ion flow tube mass spectrometry (SIFT‐MS). Garlic was also blended with water (control), polyphenol oxidase (PPO), rosemarinic acid, quercetin or catechin, and the volatiles in the headspace analyzed from 3 to 40 min by SIFT‐MS. Raw apple, raw lettuce, and mint leaves significantly decreased all of the garlic breath volatiles in vivo. The proposed mechanism is enzymatic deodorization where volatiles react with phenolic compounds. Apple juice and mint juice also had a deodorizing effect on most of the garlic volatiles but were generally not as effective as the raw food, probably because the juice had enzymatic activity but the phenolic compounds had already polymerized. Both heated apple and heated lettuce produced a significant reduction of diallyl disulfide and allyl mercaptan. The presence of phenolic compounds that react with the volatile compounds even in the absence of enzymes is the most likely mechanism. Green tea had no deodorizing effect on the garlic volatile compounds. Rosmarinic acid, catechin, quercetin, and PPO significantly decreased all garlic breath volatiles in vitro. Rosmarinic acid was the most effective at deodorization.     

茶树花的功能成分及相关产品研究进展

茶树花作为茶树生长发育过程中的副产物,具有丰富的保健功能成分和研究利用价值。中国是茶叶大国,茶树花资源丰富且开发潜力巨大。茶树花中存在丰富的保健功能成分,且与茶叶相似,对茶树花进行系统开发利用有助于增加茶农和茶企业的经济收益,实现“变废为宝”,进一步提高茶产业的附加值,为茶农创造更多的经济价值。本文主要是通过文献综述的方式,收集、整理并分析近几年茶树花的研究和开发利用现状。从茶树花资源的角度出发,对其生化成分的提取及近年茶树花产品开发利用现状进行综述,以期为茶树花资源的开发提供新的思路和参考,对茶产业的可持续发展具有一定的实践和经济价值。     

Changes in volatile compounds upon aging and drying in oolong tea production

BACKGROUND: Long‐term storage (aging) with periodic drying of fresh oolong tea gives rise to so‐called old oolong tea. Alteration of aroma compounds is expected when a fresh oolong tea is converted into an old one, as the two teas smell drastically different. The aim of this study was to compare the volatile compounds in fresh and old oolong teas. RESULTS: Significant differences were observed between the volatile compounds in fresh and old oolong teas. This observation suggested that long straight chains of alcohols and acids were putatively decomposed while shorter‐chain acids, their amide derivatives and many nitrogen‐containing compounds were generated during the tea conversion processes. The overall patterns of volatile compounds observed in five different preparations of old oolong tea were fundamentally identical. This consensus pattern was different from that observed in oolong tea either stored for more than 10 years without drying or prepared at relatively low temperatures and short baking time. CONCLUSION: Characteristic aroma nitrogen‐containing compounds, including N‐ethylsuccinimide, 2‐acetylpyrrole, 2‐formylpyrrole and 3‐pyridinol, were consistently found in the examined old oolong teas. These compounds might be regarded as typical constituents at least for a certain kind of old oolong tea. Copyright © 2010 Society of Chemical Industry     

重庆地区主要茶树栽培品种生化特性分析

以27 个重庆地区主要茶树栽培品种的1 芽2 叶鲜叶为供试材料,通过测定各品种的主要生化成分,分析各品种在重庆地区与原产地之间的差异性,及比较各品种在重庆的表现特性;并采用相关性、多维尺度和聚类分析探究27 个品种在重庆地区的表现特性及其适制性。结果表明：供试材料在重庆地区表现为氨基酸的变化率最大,咖啡碱和茶多酚含量的变化率次之;表儿茶素没食子酸酯、儿茶素、表没食子儿茶素含量和儿茶素品质指数,以及多酚氧化酶（PPO）和过氧化物酶（POD）活性的变异性较大;茶多酚与氨基酸含量呈负相关性,咖啡碱与茶多酚含量、儿茶素含量和酚氨比呈极显著和显著正相关,PPO和POD活性呈极显著正相关性;通过多维尺度和聚类分析可将供试品种分成4 大类群,且每个类群的茶类适制性都有一定的差异性。     

红茶萎凋处理温湿度条件对PPO、POD及茶多酚和色素的影响

通过控制不同温湿度进行萎凋,然后在较理想的揉捻、发酵、干燥条件下制成红茶。检测加工期间多酚氧化酶(PPO)、过氧化物酶(POD)活性以及茶多酚、茶色素含量。研究表明:不同温湿度处理萎凋期间PPO活性与萎凋叶含水量呈显著的正相关,相对湿度(RH)70%萎凋制得的红茶茶黄素含量最高,红茶发酵过程中茶多酚的氧化形成茶色素类与PPO、POD活性关系紧密,并且萎凋叶POD活性与成品茶茶黄素和茶红素含量呈极显著的负相关。      

多酚氧化酶对普洱茶渥堆发酵过程中品质变化的影响

本文探讨了添加不同浓度多酚氧化酶对渥堆发酵过程中普洱茶理化指标和感官品质的影响。实验结果表明:经不同浓度多酚氧化酶处理后的普洱茶,其部分品质成分在渥堆发酵过程中的变化差异显著(p0.01),与对照组相比,实验组的渥堆平均温度相对较高,茶多酚、儿茶素指标变化速度加快,且与多酚氧化酶的添加量呈负相关;茶黄素发酵前期生成速度加快,后期转化速度提高;茶褐素有较快积累。加入多酚氧化酶的普洱茶发酵18d后,在外形、汤色、滋味、香气、叶底等感官品质上基本具有熟茶的品质,其中1.6U/g、3.2U/g多酚氧化酶组可达到陈年普洱汤色红浓明亮,滋味醇厚回甘,香气浓郁的品质特点。本研究为缩短发酵周期,改善普洱茶品质提供新的思路和方法。