动力学机理汇总.doc

磷脂酶D催化合成磷脂酰丝氨酸反应抑制动力学的研究 摘 要：磷脂酶D（PLD）催化磷脂酰胆碱（PC）磷脂酰基转移合成磷脂酰（P）的反应在相体系中进行是目最为广泛的一种方式，但其机理尚不明确。本文在PH 5.5，温度2℃，200 rpm/min的恒温摇床中，进行不同浓度PC在PLD催化下发生的转磷脂酰基反应，发现随着PC浓度的增加，PC的转化率呈下降趋势。为验证PC转化率的下降是由于生成胆碱浓度的增高所影响的，继而在同PC浓度下分别加入3、6mM的抑制剂（胆碱）进行转磷脂酰基反应，探讨胆碱在PLD催化下的抑制机理实验结果很好的符合了所提假设，关键字：磷脂酰丝氨酸；磷脂酰胆碱；磷脂酶D；转磷脂酰基反应 Inhibition kinetics of conversion of phosphatidycholine to phosphatidylserine by phospholipase D Abstract: The synthetic reaction of phosphatidyserine from phosphatidylcholine(PC) catalyzed by phospholipase D(PLD) was carried in oil-water two phase system. It is the most extensive widely-used way for the preparation of PS, but the reaction mechanism of transphosphatidylation and hydrolysis by PLD remains unclear. The conversion of PC to PS was carried out at a Constant temperature shaking table with the speed of 200 rpm/min, pH 5.5 and 28℃, with an increase in the PC concentration, the conversion of the PC and the yield of the PS all decreased. The effect of the high choline concentration for the decrease was investigated－by the addition of 3、6mM depressor (choline) in the transphosphatidylation system with different concentration of PC,respectively. The inhibitory mechanisms of choline in PLD catalytic reaction was explored, then we described two reaction models which are free inhibition intrinsic kinetic model and inhibition intrinsic kinetic model, The experimental data agreed well with the simulation results, these model may serve as a predictor for the industrialized production of the PS. Key words: phosphatidylserine; transphosphatidylation; kinetic model; inhibition; 磷脂酰丝氨酸（PS）是一种体内分布广而功能独特的稀有磷脂，研究表明，PS能够增强神经递质效率，促进脑部信息传递，激发大脑活化状态促进用脑疲劳的恢复、改善记忆力预防老年痴呆。PS已成为重要的保健品，市场需求不断扩大。然而，天然磷脂中PS含量很低，利用磷脂酶D（PLD）催化的磷脂酰基转移反应由廉价的大豆磷脂（磷脂酰胆碱，PC）合成PS是受到研究人员广泛重视的工艺。 典型PLD催化的磷脂酰基转移反应是水-有机溶媒双液相体系，尽管在大大过量的磷脂酰基受体L-丝氨酸存在下，产物收率仍难于进一步提高，这不仅增加了原料、酶的消耗，也影响了产品PS的纯度。为了掌握磷脂酰基转移反应的规律，需要研究磷脂酶D催化磷脂合成PS过程的动力学行为Ogino等人磷脂酶D磷脂酰基转移反应乙醇胺抑制Juneja等Damnjanovi? J等总结了磷脂酰基转移反应过程的受体效应，认为受体的浓度应该足够高以确保高效，并有效抑