漆酶纯化及部分性质研究

STUDY ON PURIFICATION AND PART OF CHARACTERS FOR LACCASE

目的:研究三色革裥菌(Lenzites tricolor)所产漆酶的纯化及其部分酶学特性。方法:利用盐析、层析等方法纯化漆酶;测定漆酶的活力及不同金属离子对其的影响。结果:培养第6d时生长最快,漆酶活力最强。层析缓冲体系应选择:缓冲液Na2HPO4-C6H8O7;pH值5.0;反荷离子强度范围0.4～0.5mol/L较合适。漆酶的Km值约为60.4μmol/L。Na+、Ba2+对漆酶活力无明显影响,Cu2+、Mn2+、Zn2+、Mg2+对漆酶有不同的抑制作用,而Ca2+对漆酶有明显的激活作用。经75%(NH4)2SO4盐析、Sephadex G-150层析后得到较好的纯化漆酶。结论:三色革裥菌所产漆酶经盐析、层析得到较好的纯化效果,不同金属离子对漆酶活力具有不同的影响。

Objective： Study on the purification of laccase from Lenzites tricolor and its part of characters in enzymology. Methods： Use the ways of salting out and chromatography to purify laccase ; measure the activity of laccase and its effect by different metal ions. Results：It grew fastest and had strongest activity of laccase when cultured at 6d. It should choose proper chromatography system： buffer Na2HPO4 - C6H8O7 ; pH 5.0 ; anti - charge ionic strength range 0.4 - 0.5mol/L. Km of laccase is about 60.4umol/L. There is no obvious influence by Na^＋ ,Ba^2＋ to laccase. Cu^2 ＋ ,Mn^2＋ ,Zn^2＋ ,Mg^2＋ have different inhibitory ability to laccase, but Ca^2＋ has apparent activity. It gets a pretty result according to salting out with 75% （NH4 ）2SO4 and Sephadex G - 150. Conclusion：It has a good purified result according to the way of salting out and chromatography for laccase from Lenzites tricolor. Different metal ions have different effect on activity of laccase.