CN116103113A - 一种茶酒组合物及其制备方法和应用 - Google Patents
一种茶酒组合物及其制备方法和应用 Download PDFInfo
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Abstract
本发明涉及一种茶酒组合物及其制备方法和应用,本发明所得产品标准容易制定,产品质量容易控制,产品效果稳定。使该产品具有稳定的生态养生、延缓衰老、养肝护肝、提高免疫力等多种功效。
Description
技术领域
本发明属于食品领域,特别涉及一种茶酒组合物及其制备方法和应用。
背景技术
现代医学研究表明,人体的衰老与疾病的发生与细胞内自由基的增加密切相关。这是因为自由基含有未配对电子,状态极不稳定,为了保持其稳定状态,会从邻近的分子(包括脂肪、蛋白质和DNA)上夺取电子。如此一来,邻近的分子又变成一个新的不稳定自由基,再去进攻其它细胞或者器官。在这样的连锁反应中,自由基产生过多或是清除过慢会导致细胞的结构破坏、功能丧失、基因突变,甚至死亡(Harman 1956;寇小红2008c;袁勇等2018)。许多条件(炎症、饮食不平衡、热应激、高代谢负荷、呼吸系统疾病和寄生虫)可导致活性氧代谢产物(ROS)的形成。ROS导致蛋白质氧化,包括自由基物种,如超氧化物(·O2)、羟基(OH·)、过氧基(·RO2)、烷氧基(RO·)、氢过氧基(HO2),以及非自由基物种,如过氧化氢(H2O2)、次氯酸(HOCl)、臭氧(O3)、单线态氧(1O2)和过氧亚硝酸盐(ONOO-)。
而这些氧化会对生命不同层次的活动产生负面影响,人们对由于机体细胞内氧化应激而产生健康问题的分子机制已有了较深入了解。衰老的自由基学说认为,退行性衰老在很大程度上可以归因于ROS的累积效应92。衰老与细胞内活性氧(ROS)升高和端粒酶逆转录酶活性丧失有关。H2O2以Src家族激酶依赖的方式诱导端粒酶逆转录酶(TERT)核输出到胞浆中(Haendeler,2004)。当活性氧的浓度超过细胞的抗氧化能力时,细胞就会产生氧化应激。氧化应激导致DNA损伤,增强脂质过氧化,蛋白质损伤和细胞死亡(da Silva,2018)。
当活性氧的浓度超过细胞的抗氧化能力时,细胞就会产生氧化应激。氧化应激导致DNA损伤,增强脂质过氧化,蛋白质损伤和细胞死亡{da Silva,2018#3}。蛋白质在生物系统中含量丰富,而且反应的速率常数很高,是生物体内最重要的分子,其功能正常是保证机体健康的基础。但它们是细胞内氧化剂的主要作用目标,一些自由基和非自由基氧化剂(如单线态氧和次氯酸)的动力学数据与消耗细胞内产生的大部分这些物种的蛋白质一致。细胞内氧化剂会引起广泛的、相对非特异性的损伤,氧化可以发生在蛋白质主链和氨基酸侧链上。这种氧化引起的蛋白质损伤大部分是不可修复的,并对蛋白质结构和功能造成有害后果;然而,蛋氨酸亚砜的形成在某些情况下是可以逆转的。氧化蛋白的主要命运是通过蛋白体和溶酶体途径进行分解代谢,但有些物质似乎降解较差,并在细胞内积累。这种受损物质的积累可能会导致一系列人类疾病。
氧化会对生物最重要分子—蛋白肽连及氨基酸分子产生不同程度的变化。蛋白质氧化可导致酶活性丧失或增加,蛋白功能和蛋白酶抑制剂活性丧失,蛋白聚集,蛋白水解敏感性增强/降低,细胞摄取异常,基因转录修饰,免疫原性增强。另一方面,氧化应激是免疫衰老和炎症的关键因素。心脑血管堵塞引起的中风和心肌梗死等动脉粥样硬化性疾病对人类健康威胁很大,是导致死亡率最高。并且近年来患有人群有年轻化趋势。
发明内容
本发明所要解决的技术问题是提供一种茶酒组合物及其制备方法和应用,克服现有技术所制茶酒抗氧化能力低,产品不稳定等缺陷。
本发明的一种茶酒组合物,按重量份数,组分包括:茶叶4-10份、基酒10-60份、肽0.005-0.2份。
优选地,所述茶叶的直径为50-200μm;所述茶叶为普洱茶、红茶、绿茶、黑茶、白茶等中的一种或几种。
优选地,所述肽来源为谷物醇溶蛋白片段。
优选地,所述肽序列为
GSP1:YQYQLPSYRTNPCGVSAAIPPYY;如SEQ ID No.1所示;
GSP2:PYNQFSLMNPALQ;SEQ ID No.2所示;
GSP3:MKLVLVVLAFIALVSSVSCTQTGGCSCGQQQSHEQQHHPQ;如SEQ ID No.3所示;
GSP4:QQPISQQQQQQQQQQQKQQQQQQQQQQILQQILQQQLIPCRDVVLQQHSIAYGSSQVLQQSTYQLVQQLCCQQLWQIPEQSRCQAIHNVVHAIILHQQQQQQQQQQQKQ;如SEQ ID No.4所示;
GSP5:IHSVVHSIIMQQQQQQQQQQ如SEQ ID No.5所示中的一种或几种。
优选地,所述肽通过将谷物(如小麦、玉米、水稻、大豆种子中的一种或几种),经过蛋白酶解(如木瓜蛋白酶、菠萝蛋白酶、胰蛋白酶中的一种或几种)等电点沉淀后获得。
优选地,按重量份数,组分包括:茶叶5-10份、基酒10-60份、肽0.05-0.2份。
优选地,所述基酒的体积百分浓度为10-60%;所述基酒为食用酒;所述食用酒为白酒、红酒、黄酒、米酒中的一种或几种;所述茶酒中还含有香精。
本发明的一种所述茶酒组合物的制备方法,包括:
(1)将干燥茶叶粉碎,置于乙醇溶液中,微波处理、超临界萃取,得到茶提取物;
(2)将步骤(1)中的茶提取物、基酒混合,熟化后加入肽,混合,得到茶酒组合物。上述制备方法的优选方式如下:
所述步骤(1)中乙醇溶液为基酒,其中基酒为体积百分浓度为10-60%的酒。
所述步骤(1)中茶叶和乙醇溶液的比例1g︰3-5mL。
所述步骤(1)中微波处理为温度30-40℃,微波功率为300-500W,处理时间10-15min;所述超临界萃取为30-40℃,压力20-40MPa,萃取1-3h。
所述步骤(2)中熟化为35-65℃熟化48-120h;步骤(2)中基酒10-50份。
所述步骤(2)中加入食用香精。
本发明的一种所述茶酒组合物在食品中的应用。
有益效果
本发明不需要传统茶酒要茶叶发酵这一复杂过程,发酵工艺复杂,难以控制,同时发酵所用微生物会破坏茶叶中抗氧化活性物质,引起产品抗氧化能力下降。
本发明制备茶酒通过微波结合超临界萃取的工艺过程,同进结合醇溶蛋白肽段,可以稳定更多茶有效成份,而不需要在其中另加入茶多酚,就可达到很高的抗氧化效果。
本发明所得产品标准容易制定,产品质量容易控制,产品效果稳定。使用专利工艺生产的茶酒,活性物质不容易沉淀析出,使该产品具有稳定的生态养生、延缓衰老、养肝护肝、提高免疫力等多种功效。
本发明开发出的来自醇溶蛋白的短肽,可与茶活性通过输水作用及氢键稳定茶酒。本发明所开发的肽来自醇溶蛋白的部分酶解肽段,分子小,对茶多酚活性影响小。其显著特点是具有疏水端和亲水端。疏水端有利于和茶多酚等活性成分结合,亲水端极性有利于溶入乙醇的水溶液中。而用其它肽段就不能表现出此功能。
本发明所用得肽,为醇溶蛋白肽段。其所含的谷氨酰胺可以定向刺激人体胃肠道的肌肉蛋白和糖原的合成,可以大大提高人体胃黏膜的生成,减少酒精对胃的刺激,对解决顽固性胃炎胃溃疡等胃肠疾病最有效的天然蛋白。
本发明所得产品标准容易制定,产品质量容易控制,产品效果稳定。使该产品具有稳定的生态养生、延缓衰老、养肝护肝、提高免疫力等多种功效。
附图说明
图1为实施例2中ABTS法测定该发明所得茶酒清除氧自由基能力。Control代表对照,是不加茶提取物的酒产品。1%表示该茶酒稀释100倍氧自由基清除效果。10%表示该茶酒稀释10倍氧自由基清除效果。颜色越深,清除能力差。清除能力越强,颜色越浅。
图2为茶酒制备流程图。
图3中(A)为PST1疏水性分析(B)为PST1带电性分析;
图4中(A)为PST2疏水性分析(B)为PST2带电性分析;
图5中(A)为PST3疏水性分析(B)为PST3带电性分析;
图6中(A)为PST4疏水性分析(B)为PST4带电性分析;
图7中(A)为PST5疏水性分析(B)为PST5带电性分析;
图8中(A)为对照样疏水性分析(B)为对照样带电性分析。
具体实施方式
下面结合具体实施例,进一步阐述本发明。应理解,这些实施例仅用于说明本发明而不用于限制本发明的范围。此外应理解,在阅读了本发明讲授的内容之后,本领域技术人员可以对本发明作各种改动或修改,这些等价形式同样落于本申请所附权利要求书所限定的范围。
一、测试标准和方法
测定方法见下:
1.还原力测定:
分别于试管中准确加入0.1,0.2,0.3mL的茶酒样品,加入浓度0.2mol/L pH值6.6的磷酸盐缓冲溶液2.5mL,再加入质量分数1%的铁氰化钾溶液1.5mL,混合均匀,于50℃水浴中反应20min后急速冷却,加入质量分数10%三氯乙酸2.5mL,质量分数0.1%氯化铁1mL。用体积分数30%乙醇定容至10mL,静置10min后于波长700nm处测定吸光度A1。空白背景值以水代替氯化铁溶液,同上操作,测定吸光度A2。由公式计算其还原力,吸光度越高,说明还原性越好。
还原力=A1-A2.
式中:A1——样品组吸光度;
A2——样品本底吸光度(以等体积的蒸馏水代替三氯化铁溶液)。
2.超氧阴离子的清除率:
将茶酒样品溶液加到2支5mL试管中,一支试管加0.2mL样品液;然后在一支试管中加入0.1mM的DPPH乙醇溶液3.8mL,在另一支试管中加入3.8mL乙醇,两支试管充分摇匀后于室温下置于黑暗处孵育30min,然后于波长517nm处测定吸光度。用乙醇作空白。
按下式计算DPPH·清除率(ID):ID(%)=[1-(A1-A2)/A0]×100
式中,A1:0.2mL茶酒样液+3.8mL DPPH乙醇溶液的吸光度;
A2:0.2mL茶酒样液+3.8mL乙醇的吸光度;
A0:0.2mL乙醇+3.8mL DPPH乙醇溶液吸光度。
3.ABTS·自由基清除:
对照组:
As:0.4mL GSH+3.8mL ABTS溶液;
Ab:0.4mL样品溶剂溶液+3.8mL ABTS溶液。
样品组:
As:0.4mL样品溶液+3.8mL ABTS溶液;
Ab:0.4mL样品溶剂溶液+3.8mL ABTS溶液。
数据处理:
P=(Ab-As)/Ab×100%
P——清除率;
Ab——ABTS溶液与样品溶剂溶液混合液的吸光度;
As——ABTS溶液与待测溶液混合液的吸光度。
酒均为白酒。
实施例1
多肽制备方法如下:
取150g面粉(或玉米粉、米粉、大豆粉)于不锈钢盆中,量取蒸馏水50-100mL倒入,将面粉揉成面团,放入预先倒好200mL双蒸水的烧杯中,静置10-30min,之后反复揉洗,揉洗中均匀揉搓,尽量保持面团的完整。揉洗过程分别按照次序2×300mL,2×150mL,4×100mL,4×75mL进行,最后重复用50mL蒸馏水洗涤至没有淀粉洗出。判定终点为最后一次洗涤的溶液滴加碘液不变蓝。得到的面筋真空冷冻干燥,磨碎。按照料液比1:20-30加入30-80%的乙醇溶液,磁力搅拌1-2h,4000r/min离心20min,收集上清液。上清液加蒸馏水至沉淀达到最大值,4000r/min离心20min,收集沉淀,真空冷冻干燥,粉碎过筛,即得醇溶蛋白。所得蛋白按100:0.1-1加入木瓜蛋白酶、菠萝蛋白酶、胰蛋白酶,20-40℃放置30分钟,按五种肽等电点调制pH,沉淀所得肽。冻干或喷雾干燥。备用。
GSP1,序列YQYQLPSYRTNPCGVSAAIPPYY;
分子式:C123H175N29O35S,序列长度:23;消光系数:6400M-1cm-1;GRAVY:-0.52;平均分子量2651.93g/mol;等电点:pH 8.67。
GSP 2:PYNQFSLMNPALQ,分子式:C69H103N17O20S;序列长度:13;消光系数:1280M-1cm-1;GRAVY:-0.4;平均分子量1522.72g/mol;等电点:pH 7。
GSP 3,MKLVLVVLAFIALVSSVSCTQTGGCSCGQQQSHEQQHHPQ;
分子式:C182H296N54O57S4;序列长度:40;消光系数:0M-1cm-1;GRAVY:0.17;平均分子量4280.86g/mol;等电点:pH 7.16。
GSP 4,QQPISQQQQQ QQQQQQKQQQ QQQQQQQILQ QILQQQLIPC RDVVLQQHSIAYGSSQVLQQ STYQLVQQLC CQQLWQIPEQ SRCQAIHNVV HAIILHQQQQ QQQQQQQKQ
分子式:C553H896N180O177S4;序列长度:109;消光系数:8250M-1cm-1;GRAVY:-1.14;平均分子量13026.32g/mol;等电点:pH 8.3。
GSP 5,IHSVVHSIIM QQQQQQQQQQ;分子式:C101H166N34O33S序列长度:20;消光系数:0M-1cm-1;GRAVY:-0.96;平均分子量2416.66g/mol;等电点:pH 8.09。
实施例2
按重量份称取原料。
首先将干燥茶叶(5份,汉中仙毫)粉碎成直径100μm的颗粒,置于含52%(v/v)酒溶液中,固液比1∶3(g/ml),温度30℃,微波功率为300W,处理时间10min;之后超临界萃取,30℃,压力20MPa,萃取1h。过滤,得到茶提取有效成份,熟化后(50℃,60h)加入52%(v/v)酒15份,然后加入肽(GSP1和GSP2二者等质量混和)0.01份。搅拌混匀20分钟,分装。
将该样品54℃放置14天后,产品透亮,外观未明显改变。测定该款茶酒样品还原力,测得的还原力值为2.005。DPPH清除率保持在73.65-74.81%之间,对照样品溶剂(酒)则是在16.24-17.06%之间。ABTS法,茶酒稀释100倍后,氧自由基清除率达82%以上。
实施例3
按重量份称取原料。
首先将干燥茶叶(5份,龙井)粉碎成直径100μm的颗粒,置于含12%(v/v)酒溶液中,固液比1∶3(g/ml),温度40℃,微波功率为500W,处理时间15min;之后超临界萃取,40℃,压力40MPa,萃取3h。过滤,得到茶提取有效成份,熟化后(50℃,60h)加入12%(v/v)酒25份,肽(GSP1)0.05份;搅拌混匀20分钟,分装。
将该样品54℃放置14天后,产品透亮,外观未明显改变。测定该款茶酒样品还原力,测得的还原力值分别为2.985。DPPH清除率保持在95%以上,对照样品溶剂(酒)则是在16.24-17.06%之间。ABTS法,茶酒稀释100倍后,氧自由基清除率达93%以上。
实施例4
按重量份称取原料。
首先将干燥茶叶(5份,大红袍)粉碎成直径100μm的颗粒,置于含52%(v/v)酒溶液中,固液比1∶3(g/ml),温度35℃,功率为400W,处理时间15min;之后超临界萃取,35℃,压力30MPa,萃取2h。过滤,得到茶提取有效成份,熟化后(50℃,60h)加入52%(v/v)酒溶液15份,然后再加入肽(GSP3、GSP4,等质量混和)0.1份。搅拌混匀20分钟,分装。
将该样品54℃放置14天后,产品透亮,外观未明显改变。测定该款茶酒样品还原力,测得的还原力值分别为3.205。DPPH清除率保持在98%以上,对照样品溶剂(酒)则是在16.24-17.06%之间。ABTS法,茶酒稀释100倍后,氧自由基清除率达95%以上。
该款样品能定向刺激人体胃肠道的肌肉蛋白和糖原的合成,可以大大提高人体胃黏膜的生成,减少酒对胃的刺激,对解决顽固性胃炎胃溃疡等胃肠疾病很有效。
对比例1
按重量份称取原料。
首先将干燥茶叶(5份,大红袍)粉碎成直径100μm的颗粒,置于含52%(v/v)酒溶液中,固液比1∶3(g/ml),温度35℃,功率为400W,处理时间15min;之后超临界萃取,35℃,压力30MPa,萃取2h。过滤,得到茶提取有效成份,熟化后(50℃,60h)加入52%酒溶液15份,然后再加入对照样品肽
(DGGGDGDGSGRGGGSGSKGGDGGGSRGGSGGGGSSRGSTRGGSNRASG)0.1份。搅拌混匀20分钟,分装。
将该溶液54℃放置14天后,出现大量絮状物,并在逐渐聚集。对该溶液上述指标测定,还原力值分别为1.363。DPPH清除率为53%,对照样品溶剂(酒)则是在16.24-17.06%之间。ABTS法,茶酒稀释100倍后,氧自由基清除率为63%。产品从外观及抗氧化能力上,均下降显著。
该款样品不能定向刺激人体胃肠道的肌肉蛋白和糖原的合成,没有提高人体胃黏膜的生成。
Claims (10)
1.一种茶酒组合物,其特征在于,按重量份数,组分包括:茶叶4-10份、基酒10-60份、肽0.005-0.2份。
2.根据权利要求1所述茶酒组合物,其特征在于,所述茶叶为普洱茶、红茶、绿茶、黑茶、白茶中的一种或几种;所述茶叶的直径为50-200μm;所述肽为谷物醇溶蛋白片段。
3.根据权利要求2所述茶酒组合物,其特征在于,所述肽序列为
GSP1:YQYQLPSYRTNPCGVSAAIPPYY;
GSP2:PYNQFSLMNPALQ;
GSP3:MKLVLVVLAFIALVSSVSCTQTGGCSCGQQQSHEQQHHPQ;
GSP4:QQPISQQQQQQQQQQQKQQQQQQQQQQILQQILQQQLIPCRDVVLQQHSI
AYGSSQVLQQSTYQLVQQLCCQQLWQIPEQSRCQAIHNVVHAIILHQQQQ
QQQQQQQKQ;
GSP5:IHSVVHSIIMQQQQQQQQQQ中的一种或几种。
4.根据权利要求1所述茶酒组合物,其特征在于,所述基酒的体积百分浓度为10-60%;所述基酒为食用酒;所述食用酒为白酒、红酒、黄酒、米酒中的一种或几种;所述茶酒中还含有香精。
5.根据权利要求1所述茶酒组合物,其特征在于,按重量份数,组分包括:茶叶5-10份、基酒10-60份、肽0.05-0.2份。
6.一种权利要求1所述茶酒组合物的制备方法,包括:
(1)将干燥茶叶粉碎,置于乙醇溶液中,微波处理、超临界萃取,得到茶提取物;
(2)将步骤(1)中的茶提取物熟化后加入基酒和肽,混合,得到茶酒组合物。
7.根据权利要求6所述制备方法,其特征在于,所述步骤(1)中乙醇溶液为基酒;所述茶叶和乙醇溶液的比例为1g︰3-5mL。
8.根据权利要求6所述制备方法,其特征在于,所述步骤(1)中微波处理为温度30-40℃,微波功率为300-500W,处理时间10-15min;所述超临界萃取为30-40℃,压力20-40MPa,萃取1-3h。
9.根据权利要求6所述制备方法,其特征在于,所述步骤(2)中熟化为35-65℃熟化48-120h;
步骤(2)中基酒为10-50份;所述步骤(2)中加入食用香精。
10.一种权利要求1所述茶酒组合物在食品中的应用。
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