重组猪抑制素蛋白质中毒素残留检测及其理化稳定性  

Determination of toxin residue in recombinant pig inhibin protein and its physicochemical stability

作　　者：李辉[1] 翟颖超[1] 窦晓[1] 张甜[1] 果双双 施振旦[1] 

机构地区：[1]江苏省农业科学院畜牧研究所,江苏南京210014

出　　处：《江苏农业学报》2015年第2期362-368,共7页Jiangsu Journal of Agricultural Sciences

基　　金：江苏省农业科技自主创新基金项目[CX(13)5039];江苏省科技支撑计划项目(BE2012392)

摘　　要：为探索纯化的重组猪抑制素蛋白质毒素残留情况以及重组猪抑制素蛋白质稳定性,本研究利用鲎试剂、气相色谱和SDS-PAGE方法对重组猪抑制素蛋白质中的细菌内毒素、β-巯基乙醇的残留量及其蛋白质在不同温度、极端p H值和反复冻融条件下的稳定性进行检测。结果显示,经过3次等电点沉淀洗涤后,重组猪抑制素蛋白质中细菌内毒素和β-巯基乙醇的残留量大幅减少。通过在不同温度下的处理发现,重组猪抑制素蛋白质在4℃环境下较稳定,可以长期保存;但在37℃时,稳定性快速下降,随着温度上升而降解加速。通过极端p H值和反复冻融试验发现,重组猪抑制素蛋白质对极端p H值和反复冻融具有较好的耐受性。Tachypleus amebocyte lysate, gas chromatography and SDS-polyacrylamide gelelectrophoresis were employed for the determination of residue of bacterial endotoxin and β-mercaptoethanol in recombinant pig inhibin protein, and the stability of protein affected by different temperatures, extreme pH and multigelation, respectively. Results showed that bacterial endotoxin and β-mercaptoethanol residue in the protein were drastically decreased after isoelectric point precipitati- on washing for three times. The recombinant pig inhibin protein was quite stable at 4 ℃ for long-term preservation, started to degrade at 37 ℃ and accelerated degradation along with the rise of temperature. The protein possessed a good tolerance to the extreme pH and multigelation.

关 键 词：重组猪抑制素蛋白质 内毒素 β-巯基乙醇 残留 理化稳定性 

分 类 号：Q591.2[生物学—生物化学]