嗜硫色谱分离纯化碱性脂肪酶及其氨基酸序列测定

Purification by thiophilic chromatography and determination of N-terminal sequence of alkaline lipase

扩展青霉 (Penicilliumexpansum)FS1884所产生的碱性脂肪酶经硫酸铵沉淀、Sephacr ylS 2 0 0柱层析后 ,再经嗜硫色谱 (ThiophilicChromatography)柱层析 ,被纯化了 173.8倍 ,最终比活为 5 6 94 .9U mg。纯化后的脂肪酶达到了SDS PAGE电泳纯、PAGE电泳纯以及毛细管液相色谱 (CapillaryLiquidChromatography)纯。该脂肪酶N 端氨基酸序列测定的结果是 :ATADAAAFPDLHRAAKLSSA 。

The Alkaline lipase of Penicillium expansum FS1884 was purified 173.8-fold by ammonium sulfate precipitation, Sephacryl S-200 column and Thiophilic Chromatography .The final specific activity of the purified lipase was up to 5694.93 U/mg, which was determined to be homogeneous by SDS-PAGE, PAGE and CLC respectively. The N-terminal amino acid sequence of the enzyme was determined as follows: A T A D A A A FP D L H R A A K L S S A, which indicated the higher similarlies to the reported lipases from genus Penicillium.