CAJ | 학술논문

目的：优化缓冲液匀浆提取茶树多酚氧化酶的方法。方法以龙井43号茶树鲜叶为材料,采用缓冲液匀浆浸提法提取茶树多酚氧化酶,分别进行不同单因素浸提优化试验,测定比较提取的粗酶活性,确定适合的浸提条件。结果以鲜叶标准为一芽二叶,料液比1：2,加入内含5% PVP(w：v)、经预冷的pH5.6柠檬酸-磷酸盐缓冲液(0.15 mol/L),海砂适量,冰浴研磨,4℃条件下隔夜浸提12 h,于4℃、9000 r/min离心35 min,取上清液,为龙井43号鲜叶中多酚氧化酶缓冲液成浆浸提的最优条件。结论采用优化的浸提条件,可以制备得到高活性多酚氧化酶粗酶液,为茶树多酚氧化酶同工酶分离与酶性质研究提供了基础。
목적：우화완충액균장제취다수다분양화매적방법。방법이룡정43호다수선협위재료,채용완충액균장침제법제취다수다분양화매,분별진행불동단인소침제우화시험,측정비교제취적조매활성,학정괄합적침제조건。결과이선협표준위일아이협,료액비1：2,가입내함5% PVP(w：v)、경예랭적pH5.6저몽산-린산염완충액(0.15 mol/L),해사괄량,빙욕연마,4℃조건하격야침제12 h,우4℃、9000 r/min리심35 min,취상청액,위룡정43호선협중다분양화매완충액성장침제적최우조건。결론채용우화적침제조건,가이제비득도고활성다분양화매조매액,위다수다분양화매동공매분리여매성질연구제공료기출。
Objective To optimize the extraction method of the tea polyphenol oxidase in buffer solution homogenate.MethodsPolyphenol oxidase from fresh leaves of Longjing 43 was extracted by buffer homogenate method. Different single extraction factors were tested, and the extracted crude enzyme activity was measured and compared to determine the appropriate extraction conditions.Results The fresh leaves were of a bud with two leaves, solid (tea leaf)-liquid (extraction buffer) ratio was 1:2, and the pre-cooling citrate-phosphate buffer (pH 5.6, 0.15 mol/L) containing 5% PVP (w:v) was added, a small amount of sea sand was added, and tea leaves were ground in ice bath and extracted overnight about 12 h at 4℃. The extracted buffer was centrifuged for 35 min with 9000 r/min at 4℃, and the supernatant was obtained.Conclusion High activity of crude polyphenol oxidase solution can be prepared using the optimized extraction conditions, which benefits to the isolation and purification and the analysis of enzymatic properties for isozyme of tea polyphenol oxidase.