碧螺春茶提取物抗氧化活性研究

研究碧螺春茶提取物(Extract from Biluochun tea,EBT)体外抗氧化性作用.通过回流萃取法和冷冻干燥法,得到EBT.以Vc和槲皮素为阳性对照,对羟基自由基、超氧阴离子自由基、烷基自由基的清除率以及抑制油脂过氧化的能力进行研究.结果表明,EBT对几种自由基均有一定的清除作用,对羟基自由基、超氧阴离子自由基清除能力要比Vc和槲皮素强;对抑制油脂过氧化作用也要强于Vc.对烷基自由基自由基的清除能力要弱于Vc和槲皮素.EBT的添加量在试验范围内与其抗氧化活性呈正相关.EBT具有明显的体外抗氧化活性.     

薰衣草花茶水提物的抗氧化活性研究

研究了薰衣草花茶水提物在体外对DPPH.、OH.、O2-.的清除和抑制亚油酸过氧化作用。并比较了不同浸提温度(50、60、70、80、90℃)对薰衣草花茶水提物抗氧化活性的影响。结果表明:薰衣草花茶水提物具有良好的抗氧化活性,浸提温度越高,水提物清除O2-.作用越强;水提物清除DPPH.、OH.最佳浸提温度是60℃;水提物抑制亚油酸过氧化作用的最佳浸提温度70℃。     

Lyophilized Tea Extracts of Brickellia cavanillesii (Asteraceae): In Vitro Characterization of Biological Activity

Lyophilized Brickellia cavanillesii (LBC) tea extracts and identified chemical compounds of LBC were examined using in vitro human carcinoma liver (HepG2) cells with and without fetal bovine serum (FBS). Cells were incubated for 24 h with varying concentrations of FBS and LBC, respectively; cytotoxicity was determined spectrophotometrically using MTT (Formazan 3‐(4,5‐dimethyl‐thiazol‐2‐yl)‐2, 5‐diphenyl‐tetrazolium bromide) assay. Furthermore, the potential hypoglycemic activity of LBC tea extracts was investigated using glucose transport and metabolism proteins biomarkers. FBS (0% to 10%) increased the viability of HepG2 cells steadily with increasing concentration. Possible therapeutic effects of LBC were concentration dependent with and without FBS. The cytotoxicity of 12 identified compounds from the LBC extract suggests that the individual compounds inhibited the proliferation of HepG2 cells differentially and do not reflect the inhibition of the whole aqueous LBC. Western blot analysis of glucose facilitated transporter protein 2 (GLUT 2) expression of HepG2 cells exposed to 0 mg/mL (Control) and 0.2 mg/mL LBC for 2, 4, 6, and 24 h suggests that GLUT 2 expression was increased. Increase in GLUT 2 expression in the absence of FBS was statistically significant with time of exposure. Significant difference was observed for GLUT 2 expression between 6 and 24 h and also between 4 and 24 h at 0.2 mg/mL LBC. Results obtained indicate that LBC may exhibit antidiabetic activity. However, further studies will be necessary to clearly delineate LBC potential therapeutic benefit and biological activities in animal studies as well as other in vitro models.     

酶澄清绿茶饮料研究

研究了不同种类酶对绿茶茶汤抗沉淀效果,并采用HPLC分析了经单宁酶处理后茶汤中酯型儿茶素的变化。结果表明,单宁酶、果胶酶、木瓜蛋白酶对绿茶茶汤抗沉淀影响达极显著水平。多酚类和咖啡碱都有不同程度地减少,但没有达到显著水平。经木瓜蛋白酶处理的茶汤蛋白质含量减少,氨基酸含量同其他处理样相比有一定的增加。结合方差分析和感官审评结果表明,单宁酶对绿茶茶汤抗沉淀效果最好,单宁酶最佳作用时间为2h。在一定范围内,随着单宁酶作用时间的延长,茶汤中酯型儿茶素的总量在不断减少,茶汤的透光率随着酯型儿茶素的减少在不断增大。     

普洱茶与绿茶提取物体外抑菌效应的研究

采用滤纸片扩散法分别研究不同浓度的普洱茶和绿茶提取物对李斯特氏菌(L. monocytogenes)、猪粪链球菌(S. faecalis)、大肠杆菌(E. coli)、鼠伤寒沙门氏菌(S. typhimurium)、金黄色葡萄球菌(S. aureus)和炭疽杆菌(B. anthracis)6 株食源性微生物和致病菌的抑制效果及其最小抑菌浓度(MIC)。结果表明：普洱茶提取物在1%～7%(m/V)范围内对李斯特氏菌(L. monocytogenes)有较强的抑制作用,其MIC 为0.07mg/ml,在5%～7% 范围内对所有供试菌种均有抑制作用,对炭疽杆菌的抑制效果最好。绿茶提取物在1%～2%(m/V)范围内仅对李斯特氏菌(L. monocytogenes)有抑制作用,其MIC 为0.09mg/ml,在3% 时仅对炭疽杆菌(B. anthracis)无抑制作用,在4%～7% 范围内对所有供试菌种都有抑制作用,对猪粪链球菌抑制效果最好。     

单宁酶及协同物质对绿茶饮料稳定性的影响

研究了不同剂量的单宁酶对绿茶饮料防沉淀的作用并用HPLC分析了经单宁酶处理后茶汤中酯型儿茶素的变化,同时分别对β-环状糊精、果胶酶、木瓜蛋白酶、葡萄糖氧化酶与单宁酶协同抗沉淀作用进行了研究。结果表明,单宁酶最佳作用剂量为4.0U/g;在一定范围内,随着酶剂量的增加,茶汤中酯型儿茶素的总量在不断减少,茶汤的透光率随着酯型儿茶素的减少在不断增大;β-环状糊精、果胶酶、木瓜蛋白酶与单宁酶协同抗沉淀效果达显著水平,葡萄糖氧化酶与单宁酶协同抗沉淀有效果且差异性达到极显著水平。     

Effects of Gamma Irradiation on the Biological Activity of Green Tea Byproduct Extracts and a Comparison with Green Tea Leaf Extracts

ABSTRACT:  Green tea byproduct extract powder, a major byproduct of the green tea industry, was prepared and investigated for its biological activities after a gamma-irradiation treatment for the purpose of a brightened color. The color, total phenolic contents, antioxidative activity, tyrosinase inhibition activity, cancer cell proliferation inhibition effect, and antimutagenicity were measured and compared with those of a green tea leaf extract powder. Hunter L* values of the irradiated green leaf and byproduct extracts were increased in a comparison with the nonirradiated extracts. The contents of total phenolic compounds of the green tea leaf and byproduct extracts were 424.2 ± 11.45 and 335.5 ± 18.65 mg/g of sample, respectively. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity of the 20 ppm green tea leaf and byproduct extracts were 68.2% and 57.6%, respectively. Ferric-reducing ability of the samples showed similar results to the DPPH radical scavenging capacity. The samples showed an inhibition of tyrosinase and also a cancer cell proliferation inhibition effect. Especially, the antimutagenicities of the byproduct extracts showed a strong inhibition effect (96.20% and 57.77%) on 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp) and 2-nitrofluorene (NF), respectively. Results indicated that, although the biological activity of green tea byproduct extracts is less than that of green tea leaf extract, it may be a potential source of a functional ingredient. In addition, irradiation can be applied to improve the color and biological activity of the material.     

利用单宁酶改善绿茶滋味品质的研究

为了利用外源单宁酶改善夏茶滋味品质成分和组成,实验设计了在传统绿茶加工工艺的揉捻工序中,添加不同浓度的外源单宁酶,比较不同处理的成品茶在多酚类物质、儿茶素和氨基酸含量方面的差异,分析单宁酶提高夏季绿茶品质的效果。结果表明:单宁酶处理和对照相比,茶叶中茶多酚总量无显著性差异,但儿茶素的组成变化很大,其中表没食子儿茶素没食子酸酯(EGCG)明显下降;氨基酸总量有所增加,其中谷氨酸等一些低阈值氨基酸含量增加更为明显。这些成分变化对改善夏季绿茶滋味有积极意义,在夏茶加工中添加外源单宁酶可以在一定程度上降低苦涩味,从而提高茶汤的鲜醇度。     

高通量筛选研究茯砖茶对FXR模型的作用

将茯砖茶水提液分别用三氯甲烷、乙酸乙酯、正丁醇萃取,制得三氯甲烷层、乙酸乙酯层、正丁醇层及水层样品,茶渣用95%的乙醇提取,制得乙醇提取物,通过高通量药物筛选测定各样品对法尼酯衍生物X受体(FXR)模型的激活和抑制作用,研究茯砖茶的降脂减肥功效,结果显示茯砖茶各部份对FXR都有抑制作用,乙酸乙酯层样品激活作用较明显,从中分离制得儿茶素EGCG及ECG单体对FXR模型有很好的激活作用,证明茯砖茶有较强的降脂功能,有望开发成天然的降脂药物。茯砖茶中其他活性成分的分离有待进一步进行。     

Antibacterial Activity of Tea and Coffee: Their Extracts and Preparations

Tea, Camellia sinensis (L) O. Kuntze (Theaceae) and coffee, Coffea arabica Linn. (Rubiaceae) possess antimicrobial activity in addition to various biological properties. In this study six strains of human pathogenic bacteria were assessed for their sensitivity to aqueous extracts of tea and coffee along with their preparations i.e. supplemented with milk and sugar. Antibacterial potential of the extracts has been compared with some of the commonly employed antibiotics. Bactericidal activity of the selected extracts was assessed by viable cell count method. Both tea and coffee inhibited bacteria to a variable extent and retained their antibacterial activity even after addition of milk and sugar. Equal effectiveness of tea/coffee extracts and their preparations justifies their potential as antibacterial agents.     

单宁酶的制备及其澄清茶饮料的动力学研究

研究了微生物发酵法制备单宁酶及其对茶提取液澄清化的催化动力学。结果表明：以10％浓度的茶提取液，加入3％的单宁酶液，在35℃、pH6．0的条件下反应速率最佳，其催化动力学方程为：     

壳聚糖固定化单宁酶及其澄清绿茶汤的应用研究

以固定化单宁酶的酶活和活力回收为指标,筛选出壳聚糖海绵载体固定化单宁酶。与游离酶相比,固定化酶的最适反应pH向酸性偏移0.5,最适反应温度提高5℃;对金属离子和EDTA的稳定性均增强。固定化酶的半衰期也明显延长,4℃时半衰期由35.39h延长至324.25h,25℃时半衰期仍高达189.56h。分别以没食子酸甲酯溶液和绿茶汤为底物,经连续6批次酶促反应,固定化单宁酶的酶活保持稳定。经固定化单宁酶处理后,绿茶汤澄清度、贮存稳定性明显提高。     

Membrane Clarification of Tea Extracts

The ready-to-drink (RTD) tea beverages are becoming increasingly popular owing to the health benefits associated with tea polyphenols, but instability due to development of haze and formation of tea cream is a common problem encountered in the product. Membrane technology provides a scope to produce natural, additive-free RTD teas while overcoming the major disadvantages associated with the conventional decreaming methods. Approaches employing membranes for the clarification of extracts from black and green tea have been discussed together with their relative advantages and limitations. The article also outlines the concerns to be addressed in the future attempts employing membrane technology.     

嗜热真菌单宁酶的克隆表达及在柿子汁中的应用

对嗜热烟曲霉中的单宁酶基因afTanA在毕赤酵母中进行异源表达,分析重组酶的酶学性质及其对柿子汁抗氧化性的影响。研究表明,afTanA基因由1 767 bp碱基组成,编码588个氨基酸和一个终止密码子,存在1个Kex2酶切位点(Lys315-Arg316),其催化活性位点为Ser202、Asp455和His501。重组菌株经48 h诱导,重组AfTanA酶活力达到678U/mL。AfTanA的最适反应温度为40℃,最适pH5.0。不同金属离子浓度对酶活力的影响存在差异,在低浓度(1mmol/L)条件下,Zn2+使AfTanA酶活力提高49.65%,而Cu2+和Fe3+使该酶活力分别降低约78%和98%;在高浓度(5mmol/L)条件下,Zn2+使AfTanA酶活力降低47.12%,Cu2+和Fe3+能够完全抑制AfTanA活性。柿子汁经重组单宁酶AfTanA处理后,其抗氧化性能力显著提高,DPPH、ABTS自由基和·OH清除率分别提高15.79%,12.78%,3.4%。单宁酶AfTanA在毕赤酵母中实现了高效表达,并在果汁加工工业中具有良好的应用潜力。     

绿茶与红茶浸提液功能性成分含量和抗氧化能力的差异研究

作为全球公认的三大无酒精饮料之一,茶及其制品具有丰富的营养价值和独特的保健功效。本实验以自制的绿茶和红茶为原料,采用1,1-二苯基-2-三硝基法(DPPH)、氧自由基吸收能力法(ORAC)和细胞抗氧化法(CAA)对两种茶浸提液的抗氧化能力进行测定和比较,并研究了两种茶浸提液中各类功能性成分含量的差异。实验结果表明:两种自制茶浸提液均具有较高的抗氧化活性,但是其抗氧化能力随茶浸提液发酵程度的加深而降低(DPPH法IC50值:绿茶14.58μg DM/mL红茶24.14μg DM/mL;ORAC值:绿茶5313.92±318.75μmol TE/g DM红茶3684.14±233.59μmol TE/g DM;CAA值:绿茶26.23±0.43μmol QE/100 mg DM红茶16.41±0.17μmol QE/100 mg DM);随着发酵程度的加深,茶浸提液干物质总量降低,尤以茶多酚含量降低最为显著;茶多酚是影响茶浸提液抗氧化能力的主要功能性成分。     

海南4种茶水提物的抗氧化及抑菌活性比较

为比较海南苦丁茶、白沙绿茶、大叶种茶及鹧鸪茶4种茶水提物的抗氧化及抑菌活性,本实验通过水浸提法获得4种茶的水提物,利用分光光度法测试其体外抗氧化活性;并以5株酿酒酵母及5种致病细菌为靶标,分别测定了4种茶水提物的抗氧化活性及抑菌活性。结果表明:苦丁茶、白沙绿茶、大叶种茶及鹧鸪茶水提物的提取率分别为29.31%、23.02%、27.07%和16.18%;总多酚含量分别为(251.48±12.30)、(315.05±10.98)、(395.05±27.70)、(420.05±25.65)mg GAE/g DW;总黄酮含量分别为(691.60±19.47)、(292.60±18.36)、(524.93±1.53)、(217.60±7.00)mg RE/g DW;4种茶的总抗氧化能力结果由强到弱为:大叶种茶>鹧鸪茶>白沙绿茶>苦丁茶;特别是鹧鸪茶和大叶种茶对二苯基苦酰肼基(Diphenyl picryl hydrazinyl radical,DPPH)自由基、2'-联氨-双-3-乙基苯并噻唑啉-6-磺酸(2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid),ABTS)自由基及超氧阴离子的清除率均高于标准品芦丁和特丁基对苯二酚(Terbutyl hydroquinone,TBHQ)。同时4种茶对H₂O₂氧化应激胁迫下的5株酿酒酵母模式菌株的存活率均有提高,其中苦丁茶对基因缺陷型菌株gsh1Δ的存活率提高最明显,为30.21%。4种茶对致病细菌有较好的抑制作用,其中大叶种茶对金黄色葡萄球菌抑制效果最优,最小抑菌浓度(Minimum inhibitory concentration,MIC)为1.56 mg/mL。由此表明,4种茶水提物具有不同的抗氧化活性和抑菌活性,且多酚含量与活性之间具有较好的相关性。     

黑茶优势菌对绿茶浸提液发酵过程多酚类化合物的影响

利用从普洱茶中分离出的黑曲霉、根霉和从茯砖茶中分离出的冠突散囊菌接种云南大叶种晒青毛茶茶汤,进行单一菌株发酵,对发酵过程茶多酚类化合物的含量变化进行分析。结果表明：随着发酵时间的延长,茶多酚含量显著降低,黄酮类化合物总量在黑曲霉、冠突散囊菌作用下分别减少72%、31.76%,在根霉作用下增加了94.92%;儿茶素各组分的含量变化较大,其中表没食子儿茶素、表没食子儿茶素没食子酸酯、表儿茶素、表儿茶素没食子酸酯均呈现显著的下降趋势,而儿茶素、没食子酸在不同菌株作用下表现出不同的变化规律;茶黄素在黑曲霉、根霉作用下先增加后减少,冠突散囊菌则相反;茶红素总体呈下降趋向,茶褐素在根霉、冠突散囊菌作用下显著增加。     

Membrane Clarification of Black Tea Extracts

Microfiltration (MF; 200 and 450 nm) and ultrafiltration (UF; 25, 50, 100 and 500 kDa) membranes were assessed for the clarification of black tea extracts. Rejection of tea components including cream components increased with decrease in membrane pore size. Clarity of membrane processed extracts was excellent (～4 Nephelometric Turbidity Units) even after 30 days of refrigerated storage. UF-500 and MF membranes resulted in greater retention of original colour, besides greater recovery of tea solids including polyphenols in the clarified extract. Increase in feed concentration affected the recovery while diafiltration improved the solids recovery including polyphenols and theaflavins. Highest productivity was achieved at 2 % feed concentration followed by 0.6 %. Solids and polyphenols recoveries obtained with 0.6 % feed concentration and MF-450 membrane combination were unmatched with any other combination. The results favoured employing MF as a means of clarification of tea extracts, preferably at low feed concentration which provided greater recovery and productivity, adequately meeting the quality requirements of ready-to-drink tea beverages.     

茶树单宁酶的原核表达及酶学性质表征

基于大肠杆菌的密码子偏好性,对茶树单宁酶基因CsTanA碱基进行优化,基因优化后GC含量为51.9%,密码子适应指数为0.8,优化前后基因序列的一致性为77.8%,以pET-30a为表达载体对优化基因进行重组表达及酶学性质分析。A600 nm约0.6的重组菌株以1 mmol/L的异丙基-β-D-硫代半乳糖苷在30 ℃诱导12 h,破碎上清液重组单宁酶rCsTanA比活力为0.35 U/mg,镍柱纯化后的rCsTanA比活力为1.53 U/mg,纯化倍数约为4.4。纯化重组单宁酶rCsTanA分子质量为39 kDa,其最适温度和最适pH值分别为40 ℃和7.0。不同金属离子浓度对酶活力的影响存在差异,在低浓度（1 mmol/L）条件下,K＋增强了rCsTanA 37.28%的酶活力,而Ag＋、Cu2＋和Fe3＋使该酶活力均丧失80%以上;而在高浓度（5 mmol/L）条件下,Cu2＋表现出完全抑制rCsTanA活性的特性。表面活性剂（十二烷基硫酸钠、吐温80和十六烷基三甲基溴化铵）和极性溶剂（甲醇、乙醇、丙三醇、异丙醇及丙酮）对rCsTanA活性具有较强抑制作用。本研究不仅实现了茶树单宁酶的表达和酶学性表征,同时也丰富了单宁酶资源,为植物单宁酶的进一步应用研究提供了必要的理论支撑。