【作者】 林志明；

【导师】 姚江武；

【作者基本信息】 福建医科大学 ， 口腔临床医学， 2010， 硕士

【摘要】 目的:探讨三种巯基蛋白酶(菠萝蛋白酶、木瓜蛋白酶和无花果蛋白酶)在不同盐浓度、pH值和温度环境中水解人全唾液生物膜的效率。方法:收集非刺激性的人全唾液(whole saliva, WS),采用消散因子石英晶体微天平(QCM-D)在体外原位、实时和动态监测人非刺激性全唾液在Au石英晶体芯片形成生物膜,用菠萝蛋白酶、木瓜蛋白酶和无花果蛋白酶在不同盐浓度、pH值和温度环境中水解唾液生物膜。采用3倍频和归一化方法记录频率改变(△F)和能量消散因子改变(△D)随时间的变化。用分析软件计算膜的质量减小量(△m)和厚度减小量(△h)。通过△m和△h评价蛋白酶水解唾液生物膜的效果。结果:菠萝蛋白酶在盐浓度0.001~0.01 M时,△m和△h无明显变化,高于0.01 M时,△m和△h随着盐浓度的升高而减小。木瓜蛋白酶和无花果蛋白酶分别在盐浓度为0.005 M和0.05 M时,△m和△h最大,水解效率最高,低于或高于该盐浓度,水解效率都降低。随着pH值的升高,菠萝蛋白酶的△m和△h减小,水解效率降低,即pH 5.9>pH 6.9>pH 7.9(P<0.05);无花果蛋白酶的△m和△h增加,水解效率升高,即pH 5.9<pH 6.9<pH 7.9(P<0.05);木瓜蛋白酶则受pH值的影响较小,在pH 5.9时较高。随着温度的升高,三种蛋白酶的△m和△h都增加,水解效率升高,都在35℃时最大。结论:三种巯基蛋白酶水解人全唾液生物膜的效率都受到盐浓度、pH值和温度的影响;三种蛋白酶之间水解效率不同,无花果蛋白酶在中性和碱性环境中水解效率最高,菠萝蛋白酶则在酸性环境下水解效率最高。更多还原

【Abstract】 Objective: The aim of this paper is to investigate the efficiency of hydrolyzing human whole salivary biofilm by three thiol proteases (bromelain, papain and ficin) in different salt concentration, pH and temperatures. Methods: The whole saliva was collected by unstimulated method. The human unstimulated whole salivary biofilm was formed on the gold electrode surface by quartz crystal microbalance with dissipation (QCM-D) in real time and in situ. Bromelain, papain and ficin were used to hydrolyze the human whole salivary biofilm in different salt concentration, pH and temperatures. The normalization method and third overtone frequency were used to record the frequency shift (△F) and energy dissipation shift (△D) with variation of time. Analysis software was used to calculate the changes of the mass (△m) and the thickness (△h) of the biofilm. The efficiency of hydrolyzing human whole salivary biofilm by proteases were calculated by△m and△h. Result: The△m and△h of bromelain had no change with increasing salt concentration from 0.001 to 0.01M, but decreased with increasing salt concentration above 0.01M. Papain and ficin all had a optional salt concentration,which were 0.005M and 0.05M. The△m and△h of papain and ficin were the highest in this concentration and decreased below and above this concentration. The△m and△h of bromelain decreased with rising the pH, which was pH5.9>pH6.9>pH7.9(P<0.05). The△m and△h of ficin increased with rising the pH, which was pH5.9<pH6.9<pH7.9(P<0.05). The efficiency of papain was effected little by pH. The△m and△h of these three proteases all increased with rising the temperature. Conclusion: The efficiency of hydrolyzing human whole salivary biofilm by these three thiol proteases were effected by salt concentration, pH and temperature. The hydrolysis efficiency were difference among these proteases. The hydrolysis efficiency of ficin was the highest in the neutral and basic pH range, while bromelain has the highest efficiency in the acidic pH range.更多还原