灵芝发酵绿茶提高绿茶茶汤品质的研究
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摘要
绿茶是未经发酵的,中国产量最多,饮用最为广泛的一种茶,经杀青、揉捻、干燥,大部分白毫脱落,浸泡后为绿汤绿茶。绿茶的特点是较多的保留了鲜叶内的天然物质。其中茶多酚、咖啡碱保留鲜叶的85%以上,叶绿素保留50%左右,维生素损失也较少,从而形成了绿茶“清汤绿叶,滋味收敛性强”的特点。绿茶中保留的天然物质成分,对防衰老、防癌、抗癌、杀菌、消炎等均有特殊效果,为其他茶类所不及。目前我国的茶饮料均为调配茶,相对而言更加注重口味,茶成分的真正含量并不高。本文采用微生物液体发酵的方法,参照已有的食用菌固体发酵绿茶的报道,将液体灵芝接种到绿茶茶汤中,一方面,灵芝通过自身分泌的多酚氧化酶,降解绿茶中的茶多酚;另一方面,利用绿茶中的营养物质,将代谢产物如氨基酸、灵芝酸、灵芝多糖,排放到绿茶茶汤中,最终结果,使绿茶茶汤的酚氨比降低到合理的水平,提高低档绿茶的品质,获得集灵芝发酵液和绿茶成分于一体的新型绿茶饮料。实验主要分为三部分:灵芝和香菇在绿茶培养基中驯化生长比较;灵芝液体发酵绿茶培养基配方的优化;灵芝液体发酵绿茶发酵条件的优化。最后,测定了灵芝发酵液的粘度变化,确定70小时左右,代谢产物积累最多,引起粘度变化最明显。
(1)将3g/100mL的绿茶茶汤按25%、50%、75%、100%的浓度添加到灵芝培养基中,固体培养时,用直径法观察生长情况;液体培养时,测量茶多酚降解情况。同时,按照相同的方法驯化香菇,在相同的驯化梯度下,对比灵芝和香菇的增长量和降解茶多酚的情况,确定香菇不适合绿茶的生长环境,灵芝能很好的适应。
(2)培养基配方的优化主要包括最佳氮源的筛选、最佳碳源的筛选、最佳C/N的确定和最佳培养基配方的确定。将驯化好的固体灵芝接种到一级种培养基中,培养4-7天后,按10%的接种量,接种到二级种筛选培养基中。按照绿茶饮料综合评价指标:绿茶饮料中茶多酚和氨基酸含量都高,且酚氨比低时,绿茶饮料滋味鲜爽、回味甘甜。单因素实验确定最佳氮源为尿素,最佳碳源为蔗糖,最佳C/N为: 20:1,正交实验确定最佳培养基配方为:蔗糖3.3%,尿素0.15%,KH2PO4 0.15%,MgSO4·7H2O 0.05%。
(3)灵芝液体发酵受到环境条件的影响。主要有温度、pH、通氧量、接种量、发酵时间等。通过单因素实验,按照绿茶茶汤品质综合评价指标,确定灵芝液体发酵绿茶的最适温度为:23℃,最适pH为4.1,最适摇床转速为110r/min,最适接种量为:10%,最适发酵时间为:84h。
Green tea is a kind of tea which hasn’t been fermented, it has the most output in China、and is drunk widespread. Liquid green tea is produced by green removing、rolling、drying and soaking in boiling water. The feature of green tea is that it preserves most natural substances in fresh leaves, which tea polyphenol and caffeine preserve 85% of fresh leaves’, chlorophyll preserves 50%, and vitamin losses little, thus the green tea has the characters of“green leaves in liquid sponge ,strong convergence of taste”. Resesearch shows that the natural substances preserved in green tea have the functions of anti-aging、preventing cancer、anticancer、sterilizing、antiphlogosising, others is inferior to it. Now the green tea in our country is mainly mixed green tea drink, it pays more attention on taste and the real effective components of tea is little. This article inoculated liquid ganoderma lucidum into green tea medium, one hand, ganoderma lucidum degrades tea polyphenol because it can secrete polyphenol oxidase, on the other hand, it uses the nutrients in green tea and metabolic ganoderic acid、ganoderma lucidum polysaccharide、amino acid into the liquid green tea. Thus it decreases the phenol ammonia ratio to a proper level, improves the quality of green tea, develops a new kind of green tea drink which has the effective components of green tea and ganoderma lucidum. The experiment mainly contains three parts:the growth comparison of mushroom and ganoderma lucidum domesticated in green tea medium; the medium ingredients optimization of ganoderma lucidum fermenting green tea; the fermentation conditions optimization of ganoderma lucidum fermenting green tea. At last, we measures viscosity change of Ganoderma lucidum liquid culture, determines that around 70h, metabolites accumulates most, thus the viscosity changes most at this time.
(1) Adding the green tea soup which has a concentration of 3g/100mLto the normal ganoderma lucidum medium,the domesticated gradient is 25%、50%、75%、100%. Use the diameter analysis method to observe ganoderma lucidum’s growing suitation when it is in solid culture; Mesure the tea polyphenol content everyday when it is in liquid medium. Meanwhile domesticate mushroom in the same way, compare their growing suitation and the degree of degrading tea polyphenol at the same domesticating gradient. At last, we determine the mushroom cann’t adapt to the green tea medium, while the ganoderma lucidum can adapt to it well.
(2) The medium ingredient optimization contains the selection of the best carbon source、the selection of the best nitrogen source、the selection of the best C/N、the determination of the best medium ingredient. Inoculate the domesticated solid ganoderma lucidum to the first-degree species medium, after 4-7days, inoculate 10% first-degree species to the second class fungus medium. According to the synthetic evaluation index of the green tea drink: when the content of the tea polyphenol and amino acid are all high, moreover the phenol ammonia ratio is low, green tea drink tastes well. Using the the single factor experiment ,we determines the best source for ganoderma lucidum fermenting green tea is sucrose、the best nitrogen source is urea、the best C/N is 20:1. By the orthogonal experiment, we determine the best medium ingredient is: sucrose3.3%, urea 0.15%, KH2PO4 0.15%,MgSO4·7H2O 0.05%.
(3) Ganoderma lucidum fermenting green tea is affected by environmental factors, such as temperature、pH、oxygen quantity、inoculation amount、fermentation time. According to the synthetic evaluation index of green tea drink, we determine the optimum temperature is 23℃、the optimum pH is 4.1、the optimum rotation speed is 110r/min、the optimum inoculation amount is 10%、the optimum fermentation time is 84h.
(1)将3g/100mL的绿茶茶汤按25%、50%、75%、100%的浓度添加到灵芝培养基中,固体培养时,用直径法观察生长情况;液体培养时,测量茶多酚降解情况。同时,按照相同的方法驯化香菇,在相同的驯化梯度下,对比灵芝和香菇的增长量和降解茶多酚的情况,确定香菇不适合绿茶的生长环境,灵芝能很好的适应。
(2)培养基配方的优化主要包括最佳氮源的筛选、最佳碳源的筛选、最佳C/N的确定和最佳培养基配方的确定。将驯化好的固体灵芝接种到一级种培养基中,培养4-7天后,按10%的接种量,接种到二级种筛选培养基中。按照绿茶饮料综合评价指标:绿茶饮料中茶多酚和氨基酸含量都高,且酚氨比低时,绿茶饮料滋味鲜爽、回味甘甜。单因素实验确定最佳氮源为尿素,最佳碳源为蔗糖,最佳C/N为: 20:1,正交实验确定最佳培养基配方为:蔗糖3.3%,尿素0.15%,KH2PO4 0.15%,MgSO4·7H2O 0.05%。
(3)灵芝液体发酵受到环境条件的影响。主要有温度、pH、通氧量、接种量、发酵时间等。通过单因素实验,按照绿茶茶汤品质综合评价指标,确定灵芝液体发酵绿茶的最适温度为:23℃,最适pH为4.1,最适摇床转速为110r/min,最适接种量为:10%,最适发酵时间为:84h。
Green tea is a kind of tea which hasn’t been fermented, it has the most output in China、and is drunk widespread. Liquid green tea is produced by green removing、rolling、drying and soaking in boiling water. The feature of green tea is that it preserves most natural substances in fresh leaves, which tea polyphenol and caffeine preserve 85% of fresh leaves’, chlorophyll preserves 50%, and vitamin losses little, thus the green tea has the characters of“green leaves in liquid sponge ,strong convergence of taste”. Resesearch shows that the natural substances preserved in green tea have the functions of anti-aging、preventing cancer、anticancer、sterilizing、antiphlogosising, others is inferior to it. Now the green tea in our country is mainly mixed green tea drink, it pays more attention on taste and the real effective components of tea is little. This article inoculated liquid ganoderma lucidum into green tea medium, one hand, ganoderma lucidum degrades tea polyphenol because it can secrete polyphenol oxidase, on the other hand, it uses the nutrients in green tea and metabolic ganoderic acid、ganoderma lucidum polysaccharide、amino acid into the liquid green tea. Thus it decreases the phenol ammonia ratio to a proper level, improves the quality of green tea, develops a new kind of green tea drink which has the effective components of green tea and ganoderma lucidum. The experiment mainly contains three parts:the growth comparison of mushroom and ganoderma lucidum domesticated in green tea medium; the medium ingredients optimization of ganoderma lucidum fermenting green tea; the fermentation conditions optimization of ganoderma lucidum fermenting green tea. At last, we measures viscosity change of Ganoderma lucidum liquid culture, determines that around 70h, metabolites accumulates most, thus the viscosity changes most at this time.
(1) Adding the green tea soup which has a concentration of 3g/100mLto the normal ganoderma lucidum medium,the domesticated gradient is 25%、50%、75%、100%. Use the diameter analysis method to observe ganoderma lucidum’s growing suitation when it is in solid culture; Mesure the tea polyphenol content everyday when it is in liquid medium. Meanwhile domesticate mushroom in the same way, compare their growing suitation and the degree of degrading tea polyphenol at the same domesticating gradient. At last, we determine the mushroom cann’t adapt to the green tea medium, while the ganoderma lucidum can adapt to it well.
(2) The medium ingredient optimization contains the selection of the best carbon source、the selection of the best nitrogen source、the selection of the best C/N、the determination of the best medium ingredient. Inoculate the domesticated solid ganoderma lucidum to the first-degree species medium, after 4-7days, inoculate 10% first-degree species to the second class fungus medium. According to the synthetic evaluation index of the green tea drink: when the content of the tea polyphenol and amino acid are all high, moreover the phenol ammonia ratio is low, green tea drink tastes well. Using the the single factor experiment ,we determines the best source for ganoderma lucidum fermenting green tea is sucrose、the best nitrogen source is urea、the best C/N is 20:1. By the orthogonal experiment, we determine the best medium ingredient is: sucrose3.3%, urea 0.15%, KH2PO4 0.15%,MgSO4·7H2O 0.05%.
(3) Ganoderma lucidum fermenting green tea is affected by environmental factors, such as temperature、pH、oxygen quantity、inoculation amount、fermentation time. According to the synthetic evaluation index of green tea drink, we determine the optimum temperature is 23℃、the optimum pH is 4.1、the optimum rotation speed is 110r/min、the optimum inoculation amount is 10%、the optimum fermentation time is 84h.
引文
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