家蝇蛹中多酚氧化酶的分离纯化及抑菌活性的研究

Isolation and Purification of Phenoloxidase from Musca Domestica and Its Activity of Inhibiting Proliferation of Bacteria

采用缓冲液浸提、超滤、亲和层析和高效液相色谱4种方法,从家蝇蛹中分离纯化一种D-半乳糖专一性的多酚氧化酶(PO),并对其抑菌活性进行了研究.在SDS-PGAGE中呈现出两个条带,其中表观分子质量为78ku的条带为目的蛋白;经过高效液相色谱分析,呈现出了分别和SDS-PAGE结果中相对应的两个峰,其中第一个峰为要纯化的目的蛋白;目的蛋白经过肽指纹图谱检测为PO,精确分子质量为79ku.1mL纯化的PO在1g/L的质量浓度下对1mL稀释后的大肠埃希菌、枯草芽胞杆菌和伤寒沙门菌(106 mL-1)的抑制率分别为43.2%、38.7%和50.1%.

A D-galactose specific phenoloxidise （ PO ） was purified from Musca domestica pupue and its antibacterial activity was studied. The purification procedures included： crude extract in buffer, ultra-filtration, affinity-chromatography and HPLC. By SDS-PAGE, with or without reduction, purified PO yields two bands with a molecular weight of 78 ku. By HPLC, purified PO shows two peaks. In MALDI-TOF-MS, the precise molecular weight is identified as 79 ku. Purified PO shows significant antibacteria activity against Escherichia coli ATCC25922, Bacillus subtilis ATCC9372, Salmonella typhi- murium CCTCC94010 with the inhibiting activity of 43.2 %, 38.7 %, 50.1%, respectively.