ZtNH_2-HCl和剪切力对茶叶细胞悬浮培养中茶氨酸合成的影响

Effects of ZtNH_2-HCl and Shearing Force on Theanine Biosynthesis in Tea-Cell Suspension Culture

以婺源绿茶嫩叶愈伤组织为材料,在采用摇床悬浮培养与发酵罐悬浮培养,分析了茶氨酸合成前体盐酸乙胺(ZtNH2-HCl)不同的添加方式和剪切力对培养细胞增长量和茶氨酸合成量的影响。结果显示,发酵罐放大培养取得了与摇床悬浮培养类似的效果;在一个培养周期中,培养细胞茶氨酸积累高峰出现在第20～22天;发酵罐大规模培养时采用桨叶式搅拌器(低剪切力)细胞增长量和茶氨酸合成量优于标准板搅拌器;添加盐酸乙胺可大幅度提高茶氨酸积累量,先加入一定量盐酸乙胺再每天进行少量补充,茶氨酸合成量比一次性加入的效果要好。

The suspension cultures on the callus obtained from tender leaves of the Wuyuan tea were conducted on a shaking table and in a fermenter and then effects of different adding modes and shearing forces of ZtNH_2-HCl,the synthetic precursor of theanine,on the growth of tea cells and the amount of synthesized theanine were analyzed.The results indicated that the amplified culture carried out in a fermenter performed as similarly as the suspension culture carried out on a shaking table;in a culture period,the theanine accumulation peaked between the twentieth day and the twenty-second day;the culture carried out in a fermenter equipped with a balde agitator (low shearing force) presented a higher increment of cellular growth and a higher amount of synthesized theanine than the culture carried with an on-gauge plate agitator;adding ZtNH_2-HCl could greatly raise the amount of accumulated theanine,with the multi-addition of ZtNH_2-HCl performing better than the one-time addition of ZtNH_2-HCl at a specific amount at first and then at a smaller amounts.