CAJ | 학술논문

目的：探讨不同环境因素和处理方法对发形霞水母（ Cyanea capillata）触手提取物（ tentacle extract ， TE）蛋白稳定性及其溶血活性的影响。方法结合蛋白浓度测定、溶血活性检测和SDS-PAGE分析等方法研究不同环境因素和处理方法对TE蛋白稳定性及其溶血活性的影响。结果 TE溶血活性呈明显的剂量依赖关系，其半数溶血分数HU50＝226μg／ml；40℃水浴1 h，能去除TE中大量杂蛋白，但仍保持显著溶血活性；TE在4℃放置28 d，其溶血活性保持稳定，25℃下3 d内其溶血活性变化不明显；pH值对TE溶血活性的影响呈钟形曲线，在pH 6．0～11．0之间活性相对稳定，pH 8．0是TE保持溶血活性的最优pH条件；不同缓冲液对TE稳定性及其溶血活性影响显著，浓度大于26％的硫酸铵溶液对TE溶血蛋白具有良好的盐析作用。结论40℃水浴1 h预处理可显著减少TE中非活性蛋白组分，并降低样品黏性；4℃和pH 8．0是TE保持蛋白稳定性和溶血活性的最适条件；浓度大于26％的硫酸铵盐析有利于溶血蛋白组分富集。
목적：탐토불동배경인소화처리방법대발형하수모（ Cyanea capillata）촉수제취물（ tentacle extract ， TE）단백은정성급기용혈활성적영향。방법결합단백농도측정、용혈활성검측화SDS-PAGE분석등방법연구불동배경인소화처리방법대TE단백은정성급기용혈활성적영향。결과 TE용혈활성정명현적제량의뢰관계，기반수용혈분수HU50＝226μg／ml；40℃수욕1 h，능거제TE중대량잡단백，단잉보지현저용혈활성；TE재4℃방치28 d，기용혈활성보지은정，25℃하3 d내기용혈활성변화불명현；pH치대TE용혈활성적영향정종형곡선，재pH 6．0～11．0지간활성상대은정，pH 8．0시TE보지용혈활성적최우pH조건；불동완충액대TE은정성급기용혈활성영향현저，농도대우26％적류산안용액대TE용혈단백구유량호적염석작용。결론40℃수욕1 h예처리가현저감소TE중비활성단백조분，병강저양품점성；4℃화pH 8．0시TE보지단백은정성화용혈활성적최괄조건；농도대우26％적류산안염석유리우용혈단백조분부집。
Objective To investigate the influencing factors of the protein stability and hemolytic activity of tentacle extract ( TE) from the jellyfish Cyanea capillata .Methods Effects of various factors and treatments on the protein stability and hemolytic ac-tivity of TE were explored by protein detection , hemolytic assay and SDS-PAGE analysis.Results TE caused a significant and dose-dependent hemolytic effect , and the HU50 of TE against 0.5%erythrocyte suspensions from SD rats was 226μg/ml.A 40℃water bath for 1 hour could effectively remove the contaminating proteins in TE .TE retained hemolytic activity at 4℃for 28 days but it was unsta-ble when kept at 25℃over 3 days.TE was active in the range from pH 6.0 to 11.0 and the optimum pH was 8.0.Various buffer solu-tions had significantly different effects on the stability and hemolytic activity of TE , and a good salting-out effect was observed on the hemolytic protein of TE while the concentration of ammonium sulfate solutions was greater than 26%.Conclusion A 40℃water bath for 1 hour could effectively remove the contaminating proteins in TE and reduce its viscosity .The optimum conditions for maintaining stability and hemolytic activity of TE were 4℃ and pH 8.0.The salting-out effect from 26% and more ammonium sulfate solutions would be conducive to the enrichment of hemolytic protein .