摘要
经正醇抽提、硫酸铵分级沉淀、DEAE SepharoseFF和SephacrylS 200柱层系,从瘦肉型猪(PIC344)精液中提取出电泳纯碱性磷酸酶。用NBS,PMSF,TNBS,DTT及AcBr在一定条件下分别选择性的作用于PIC344AKP的Trp,Ser,Lys,His及巯基,并对酶活力的变化和吸收光谱的变化作了测定,结果表明,NBS,TNBS,PMSF,DTT的修饰能显著的抑制AKP的活力,活力的降低与修饰剂的浓度呈正相关;AcBr的修饰对AKP的活力没有明显的影响。作者认为Trp,Ser,Lys及巯基可能是AKP活力的必须基团。
Alkaline phosphatase(AKP)was isolated and purified from the sperm of pig(PIC344).The partially purified AKP was purified by saltingout, DEAESepharose FF ionexchange chromatography, and gel filtrition with Sephacryl S200.The purified enzyme showed a single band in PAGE.The enzyme was modified by NBS,PMSF,TNBS,DTT and AcBr and the changes in their activities and absorption spectra were measured. It was found that the reactions of AKP from PCI344 with NBS,PMSF,TNBS and AcBr resulted in a strong inhibition of enzyme activities.The activities of AKP decreased steadily with the increase of modifiers concentration.These results showed that Trp,Lys Ser and 2SH group were essential for the activities of AKP from PCI344.It was also shown that His was indifferent to the activities of AKP from PCI344.
出处
《西南农业学报》
CSCD
2003年第3期79-82,共4页
Southwest China Journal of Agricultural Sciences
