聚乙二醇-α-胰凝乳蛋白酶缀合物的酶活性和热稳定性
Enzymatic activity and thermal stability of PEG-alpha-chymotrypsin conjugates.
作者信息
Rodríguez-Martínez José A, Rivera-Rivera Izarys, Solá Ricardo J, Griebenow Kai
机构信息
Department of Chemistry, University of Puerto Rico-Río Piedras, PO Box 23346, San Juan, PR 00931-3346.
出版信息
Biotechnol Lett. 2009 Jun;31(6):883-7. doi: 10.1007/s10529-009-9947-y. Epub 2009 Feb 18.
Abstract
Alpha-chymotrypsin was chemically modified with methoxypoly(ethylene glycol) (PEG) of different molecular weights (700, 2,000, and 5,000 Da) and the amount of polymer attached to the enzyme was varied systematically from 1 to 9 PEG molecules per enzyme molecule. Upon PEG conjugation, enzyme catalytic turnover (k (cat)) decreased by 50% and substrate affinity was lowered as evidenced by an increase in the K (M) from 0.05 to 0.19 mM. These effects were dependent on the amount of PEG bound to the enzyme but were independent of the PEG size. In contrast, stabilization toward thermal inactivation depended on the PEG molecular weight with conjugates with the larger PEGs being more stable.
摘要
用不同分子量（700、2000和5000道尔顿）的甲氧基聚（乙二醇）（PEG）对α-胰凝乳蛋白酶进行化学修饰，每个酶分子连接的聚合物量从1到9个PEG分子系统地变化。PEG偶联后，酶催化周转率（k (cat)）降低了50%，底物亲和力降低，这由K (M) 从0.05 mM增加到0.19 mM证明。这些影响取决于与酶结合的PEG量，但与PEG大小无关。相反，对热失活的稳定性取决于PEG分子量，较大PEG的缀合物更稳定。
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