(S)-Linalyl, 2-Phenylethyl,and Benzyl Disaccharide Glycosides Isolated as Aroma Precursors from Oolong Tea Leaves

Three glycosides, 6-O-β-d-xylopyranosyl-β-d-glucopyranosides (β-primeverosides) of the aroma constituents, linalool, 2-phenylethanol, and benzyl alcohol, were isolated as aroma precursors from the tea leaves (Camellia sinensis var. sinensis cv. Shuixian and Maoxie, cultivars for oolong tea). The isolation was guided by acid or enzymatic hydrolysis, and subsequent GC and GC-MS analyses. The linalyl glycoside is the first example of naturally occurring (S)-linalyl β-primeveroside.     

trans- and cis-Linalool 3,6-Oxide 6-O-β-d-Xylopyranosyl-β-d-glucopyranosides Isolated as Aroma Precursors from Leaves for Oolong Tea

New glycosidic aroma precursors (1 and 2) of the main volatile constituents, trans- and cis-linalool 3,6-oxides (linalool oxides I and II), were isolated from oolong tea leaves (Camellia sinensis var. sinensis cv. Maoxie). The isolation was guided by an enzymatic hydrolysis with acetone powder prepared from fresh tea leaves (cv. Yabukita) followed by GC or GC-MS analyses. Chromatographic purification of hot water extracts of the tea leaves on active charcoal, Amberlite XAD-2, and Sephadex LH-20 columns as well as HPLC gave two new glycosides, trans- and cis-linalool 3,6-oxide 6-O-β-d-xylopyranosyl-β-d-glucopyra-nosides (1 and 2).     

Optical Isomers of Linalool and Linalool Oxides in Tea Aroma

The main aroma components of oolong and black tea, linalool and four diastereomers of linalool oxides (LOs), were enantioselectively isolated by capillary gas chromatography, using a column coated with an optically active liquid phase, permethylated β-cyclodextrin.

The R/S ratio varied among linalool and LOs, and among the different types of tea, the ratio for a particular compound also being different. However, the complete patterns of R/S ratio were similar in the semi-fermented and fermented teas, respectively.

Using a specific cultivar of black tea, the R/S ratio for each of the five compounds was compared in the free state in black tea with that of an aglycone of the glycoside in fresh tea leaves or in black tea. While the e.e. values of the compounds varied, those for a specific compound were similar, except for linalool, regardless of their free or combined state.

These results show that LOs are not directly transformed from linalool, but are formed enzymatically from glycoside precursors.     

cis- and trans-Linalool 3,7-Oxides and Methyl Salicylate Glycosides and (Z)-3-Hexenyl β-D-Glucopyranoside as Aroma Precursors from Tea Leaves for Oolong Tea

Two new alcoholic aroma precursors, cis- and trans-linalool 3,7-oxides 6-O-β-D-apiofuranosyl-β-D-glucopyranosides (1 and 2), as well as two already known compounds, (Z)-3-hexenyl β-D-glucopyranoside (3) and methyl salicylate 6-O-β-D-xylopyranosyl-β-D-glucopyranoside (β-primeveroside: 4), and another new monoterpendiol glycoside, 8-hydroxygeranyl β-primeveroside (5) have recently been isolated as aroma precursors in tea leaves (Camellia sinensis var. sinensis cv. Maoxie) ready for oolong tea processing.     

Volatile and Non-volatile Forms of Aroma Compounds in Tea Leaves and Their Changes due to Injury

Fresh tea leaves were homogenized in a chloroform-methanol mixture (1:1, v/v), and separated into chloroform-soluble and methanol-water-soluble fractions after addition of water. From the chloroform-soluble fraction, the volatile forms of the aroma compounds were obtained. The non-volatile forms of the aroma compounds were associated with the methanol-water-soluble fraction, and were converted to volatile forms by hydrolysis with dilute acid.

The amount of the aroma compounds in the free form, such as cis-3-pentenol, hexanol, cis-3-hexenol, trans-2-hexenol, linalool, linalool oxide (cis, 5-membered), linalool oxide (trans, 5-membered), linalool oxide (trans, 6-rnembered), linalool oxide (cis, 6-membered), nerol, geraniol, phenylmethanol, and 2-phenylethanol, markedly increased during black tea manufacture. However, those in the bound form, showed a slight decrease during the manufacture. The increases in the former were also brought about by maceration, or treatment of the tea leaves with monoiodoacetate or malonate.     

Flavor of Black Tea

In comparing the aroma concentrates from various types of black tea by the use of gas chromatography (GLC), differences of aroma pattern were recognized among these black tea of Ceylon, India, Peru, Formosa and Japan.

One of the typical differences, by which the variety would be characterized, appeared in the proportion of linalool (include its oxides) to geraniol and phenylethanol. Furthermore the ratio of the total area of peaks before and after linalool seemed to have some relation with the variety of black tea.

Also, top note of black tea aroma was compared by head space vapor analyses.     

Green tea aroma fraction reduces β-amyloid peptide-induced toxicity in Caenorhabditis elegans transfected with human β-amyloid minigene

Green tea is a popular world-wide beverage with health benefits that include preventive effects on cancer as well as cardiovascular, liver and Alzheimer’s diseases (AD). This study will examine the preventive effects on AD of a unique aroma of Japanese green tea. First, a transgenic Caenorhabditis elegans (C. elegans) CL4176 expressing human β-amyloid peptide (Aβ) was used as a model of AD. A hexane extract of processed green tea was further fractionated into volatile and non-volatile fractions, named roasty aroma and green tea aroma fractions depending on their aroma, by microscale distillation. Both hexane extract and green tea aroma fraction were found to inhibit Aβ-induced paralysis, while only green tea aroma fraction extended lifespan in CL4176. We also found that green tea aroma fraction has antioxidant activity. This paper indicates that the green tea aroma fraction is an additional component for prevention of AD.     

Preliminary Studies on Flavour Volatiles in Qimen Black Tea

Three kinds of flavor enriched black tea in the world are produced from Qimen in China, Darjeeling in India and Uva in Sri Lanka. The results of analysis from the Qimen black tea showed that there are more than twenty flavour volatiles. The essential aromatic compounds are geraniol, 2,6-di-butul-l,4-benzoquinone, 2,6-di-t-butyl-4-methyl phenol, benzenemethanol, benzenethanol, linalool and linalool oxides. In Qimen black tea the aroma acquired is produced during the processing aside from the original compounds of fresh leaves.     

真菌侵染引发的茶树内源糖苷酶基因差异表达

通过探讨顺-3-己烯醇、芳樟醇氧化物、芳樟醇、水杨酸甲酯、香叶醇、苯甲醇和苯乙醇7种茶叶游离态香气组分和糖苷类香气前体对茶炭疽病菌、茶云纹叶枯病菌、茶轮斑病菌和茶赤叶斑病菌4种致病菌的抑制作用,以及真菌侵染引发的茶树（Camellia sinensis）内源β-樱草糖苷酶、β-木糖苷酶、β-葡萄糖苷酶I、β-葡萄糖苷酶II、β-葡萄糖基转移酶基因的表达差异,结果表明：7种游离态香气组分和糖苷类香气前体对4种致病菌均有明显的抑制作用,其中香叶醇的抑制作用最强,在浓度为0.1mg.mL-1时即对茶炭疽病菌的抑制率达到100%。实时定量PCR结果显示：真菌侵染可不同程度诱导茶树内源糖苷酶和β-葡萄糖基转移酶基因的表达上调,且上调多发生于染病初期。     

茶树HMG-CoA还原酶基因全长cDNA克隆及序列分析

香气是茶叶的重要品质之一,萜类物质不仅香气好,而且沸点普遍较高,是构成茶叶香气的重要物质基础,决定着茶叶的香气品质,也可作为茶叶香型划分的依据。在植物中,倍半萜、多萜醇等通过胞质中的甲瓦龙酸(MVA)途径合成。HMG-Co A还原酶(HMGR)催化HMG-Co A(3-羟基-3-甲基戊二酸单酰辅酶A)生成甲瓦龙酸,是依赖MVA萜类合成途径的关键限速反应。为了有助于理解茶树萜类合成的分子遗传机制,通过RACE-PCR方法从茶树中克隆了一个编码HMG-Co A还原酶的c DNA全长序列(命名为Cs HMGR1),该序列由1 979 bp组成,包含一个1 722 bp的完整开放阅读框,编码573个氨基酸。其推定的编码蛋白与橡胶树、旱莲木、人参、荔枝、西洋参、丹参、罗汉果及龙眼的同源蛋白具有80%~82%的序列一致性。利用Cs HMGR1和其它物种HMGR同源蛋白的催化区域构建系统发育树,表明其属于真核生物I类HMGR家族。结构分析表明,Cs HMGR1含有两个跨膜区,推测其与其它真核生物同源蛋白类似地定位于内质网上;含有两个HMG-Co A结合位点、两个NADPH结合位点、四个保守的催化活性残基及一个磷酸化位点,说明磷酸化/去磷酸化很可能也是其活性调节的重要方式。表达分析表明,Cs HMGR1在"大叶龙"叶芽、母株叶芽及花芽都有较强的表达。其表达调控及生理活性对茶叶品质可能有重要影响,并在其功能解析的基础上,有可能作为茶叶品质鉴定及育种的一个依据。     

Isolation of a psychrotolerant Debaryomyces hansenii strain from fermented tea plant (Camellia sinensis) leaves

Abstract

A psychrotolerant, halotolerant and alkalophilic yeast was isolated from fermented leaves of Camellia sinensis Kuntze, the tea plant. The yeast strain, named Tea-Y1, was both phenotypically and genotypically identified as belonging to the species Debaryomyces hansenii. This assignment was confirmed by scanning and transmission electron microscopy. The analysis of growth curves demonstrated the ability this yeast strain to grow in a temperature range between 4°C and 28°C, with an optimum of 23°C. The ecology of this yeast in the C. sinensis phyllosphere, as well as its possible role in tea fermentation and storage, with particular reference to iced tea, are discussed.     

茶树苯丙素类/苯环型挥发性物质的生物合成和胁迫响应的研究进展

茶(Camellia sinensis)的挥发性物质不仅会影响茶香的品质,也会在茶树中发挥重要的生态学功能。除了萜类和脂肪酸衍生类挥发性物质外,苯丙素类/苯环型挥发性物质(volatile phenylpropanoids/benzenoids, VPBs)也同样具有这些方面的功能。正因为具有这些功能,茶树中的VPBs也受到了人们越来越多的重视和研究。对近年来茶中一些具代表性VPBs的研究进展进行了综述,包括2-苯乙醇、苯乙醛等VPBs的生物合成及其对光胁迫的响应;1-苯乙醇和苯乙酮的生物合成途径及其对昆虫胁迫的响应;吲哚的生物合成及其在温度和机械损伤胁迫下的响应机制。此外,也探讨了VPBs中一些非传统的生物合成途径,以及这些途径在某些胁迫条件下的响应机制。     

Photoassimilatory and Photorespiratory Behaviour of Certain Drought Tolerant and Susceptible Tea Clones

Net photosynthetic rate (P _N) in the mother leaves was higher in the drought tolerant (DT) clones of tea (Camellia sinensis) while liberation of the fixed ¹⁴C in light from the mother leaves was higher in the drought susceptible (DS) clones. The DT clones translocated more photosynthates to the crop shoots (three leaves and a bud) from the mother leaf than the DS clones. Concentrations of RuBP carboxylase (RuBPC) or oxygenase (RuBPO) had no relationship with the drought tolerant nature of tea clones but their ratio correlated with the same. DT tea clones had higher catalase activity that could scavenge the hydrogen peroxide formed in the photorespiratory pathway and thereby reduced photorespiration rate (P _R). The ratio of RuBPC/RuBPO had a positive correlation with P _N and catalase activity. Negative correlation between RuBPC/RuBPO and P _R and between catalase activity and RuBPO activity was established.     

Isolation of Z-3-Hexenal in Tea Leaves,Thea sinensis,and Synthesis Thereof

Z-3-Hexenal, a precursor in the biosynthesis of leaf alcohol and leaf aldehyde, was first isolated from fresh tea leaves, Thea sinensis and its structure was confirmed by unequivocal synthetic evidence.     

Hypotriacylglycerolemic and Antiobesity Properties of a New Fermented Tea Product Obtained by Tea-Rolling Processing of Third-Crop Green Tea (Camellia sinensis) Leaves and Loquat (Eriobotrya japonica) Leaves

We manufactured a new fermented tea by tea-rolling processing of third-crop green tea (Camellia sinensis) leaves and loquat (Eriobotrya japonica) leaves. The mixed fermented tea extract inhibited pancreatic lipase activity in vitro, and effectively suppressed postprandial hypertriacylglycerolemia in rats. Rats fed a diet containing 1% freeze-dried fermented tea extract for 4 weeks had a significantly lower liver triacylglycerol concentration and white adipose tissue weight than those fed the control diet lacking fermented tea extract. The activity of fatty acid synthase in hepatic cytosol markedly decreased in the fermented tea extract group as compared to the control group. The serum and liver triacylglycerol- and body fat-lowering effects of the mixed fermented tea extract were strong relative to the level of dietary supplementation. These results suggest that the new fermented tea product exhibited hypotriacylglycerolemic and antiobesity properties through suppression of both liver fatty acid synthesis and postprandial hypertriacylglycerolemia by inhibition of pancreatic lipase.     

A New Glucuronide Saponin from Tea Leaves (Camellia sinensis var. sinensis)

A new glucuronide saponin (1) was isolated as its methyl ester (2) from the leaves of Camellia sinensis var. sinensis. On the basis of its spectral data and the results of chemical degradation, the structure was elucidated to be 3-O-1{β-d-galactopyranosyl(l → 2)-[β-d-xylopyranosyl(1 → 2)-α-l-arabinopyranosyl(1 → 3)]-β-d-glucuronopyranosyl}-21-O-cinnamoyl-16,22–di-O-acetylbarringtogenol C.