차 유전자원의 특성평가 및 선발계통의 생리활성과 제다특성

Abstract

Tea has long been one of the most popular non alcoholic beverages worldwide for thousands of years. And tea has continually been improved, increasing popularity and palatability. Recently, the functional effects of tea have been scientifically proved and there has been increased development of functional products using tea components. It is particularly true that most people think the tea plant is the base of the all tea industry and science. But, over thousands of years, palatability has been the aim of selecting new cultivars and preserving genetic resources. Therefore, there has been a great deal of artificial selection. Therefore, the objectives of this study were to determine the catechin and functional compounds in tea leaves, and to determine there contents in tea germplasms, also, to search a bioactive effectiveness of new compounds of tea and to set a selection standard of tea cultivars by specific manufacturing and sensory taste characters for new functional tea cultivars. Collected germplasms were classified into three groups based on the sprouting time, i.e, early, mid and late sprouting time, and the ratios were 20%, 43% and 37%, respectively. There was a difference in characteristics among these collected germplasms, leaf width of those ranged from 19.8 to 75 mm, leaf length was 35.5-160.0 mm, and leaf area was 660-8,400 mm2. The concentrations of the total nitrogen, total free amino acids, and theanine were ranged 4.18-6.07%, 2.87-4.58%, and 1.64-2.66%, respectively. Also, catechin concentration showed from 50.67 to 147.67mg/g, and concentration of caffeine was 2.82-4.23%. The acetone extracts of green tea leaves have extensive absorbance at 440- 460nm and 650-660nm. Using RP-HPLC method, 11 major peaks were detected. The 11 pigments were purified using C18 open columnchromatography and semi-preparative HPLC technique, and their chemical structures were identified by spectroscopic methods. The structures of these three pigments were elucidated as lutein, chlorophyll b, a and pheophytin a, b, β-carotene and EGCG3Me. A simple, rapid and reproducible RP-HPLC method was developed for quantitative determination of these compounds. The pigments were separated on the YMC ODS AM 303 (4.6 × 250mm) column using the gradient system. Hi- functional tea cultivars were developed using the variety germplasm. The cultivar, JANGWON No. 3 contained high EGCG3Me as 4.5 mg·g-1. EGCG3Me is one of new catechin components, which is not existed in introduced cultivars and very rarely existed in collected germplasm. The cultivar, JANGWON No. 4 contained high lutein as; 553.1 ㎍·g-1. It is higher than that of the green tea cultivar 'Yabukita' by nearly 4 folds. The antioxidative activities of JANGWON No. 3 and JANGWON No. 4 were measured using the DPPH radical, hydroxyl radical, superoxide anion. The results showed that the superoxide anion activity of high lutein cultivar 'JANGWON No. 4' is higher than ascorbic acid and Yabukita at the same concentration. The in vitro anticancer effects of EGCG3Me and lutein were studied using 3-(4,5-dimethlthiazol)-2,5-diphenyltetrazoliun bromide (NTT) assay on human cancer cells (Lung cancer cell line: A549, Renal cancer cell line: ACHN, Colorectal cancer cell: HCT15, Prostate cancer line: LNCaP, Breast cancer cell line: MCF-7). The cancer cell growth inhibition activities of lutein at the concentration of 50 ㎍·mL-1 showed less than 50% compared to control in all cancer cell lines. Lutein showed inhibition of 38% (50 ㎍·mL-1), 26% (100 ㎍·mL-1), 15% (200 ㎍·mL-1) at LNCaP and 39% (50 ㎍·mL-1), 28% (100 ㎍·mL-1), 19% (200 ㎍·mL-1) at MCF-7. EGCG3Me showed inthibition of 46% (50 μ㎍·mL-1), 37.7% (100 ㎍·mL- 1), 20% (200 ㎍·mL-1) at LNCaP. EGCG3Me has more efficient inhibitory effect than control. These results suggested that the cancer cell growth inhibition activities of EGCG3Me and lutein showed a concentration dependent effect. Among different tea manufacturing methods, the non-fermented tea from the cultivar JANGWON No. 3 with high EGCG3Me content is more bitter than half-fermented tea and fermented tea of this cultivar. That is resulted due to the changing of rich catechin components into the theaflavin and thearubidin as fermentation take place. Sensory test score of fermentation in JANGWON No. 4 is reduced as the order of non-fermentation > halffermentation > fermentation.