Improved Complementary Polymer Pair System: Switching for Enzyme Activity by PEGylated Polymers
Improved Complementary Polymer Pair System: Switching for Enzyme Activity by PEGylated Polymers
复制标题
DOI：
10.1021/la2043312
复制
发表时间：
2012-03-06
期刊：
影响因子：
3.9
通讯作者：
Shiraki, Kentaro
中科院分区：
文献类型：
--
作者：
Kurinomaru, Takaaki;Tomita, Shunsuke;Shiraki, Kentaro
酶活性开关技术的发展有望扩大酶在广泛研究领域的应用。我们之前已经开发了一种互补聚合物对系统(CPPS)，它使几种酶的活性能够被一对相反电荷的聚合物控制。然而，由于阴离子α-淀粉酶与阳离子聚烯丙基胺(PAA)的络合物聚集导致酶的不可逆变性，它不能控制大而不稳定的α-淀粉酶的活性。为了解决这个问题，我们设计并合成了一种以聚乙二醇为主链的阳离子共聚物，即聚(N，N-二乙氨基乙基甲基丙烯酸乙酯)-嵌段聚乙二醇(PEAMA-b-PEG)。与PAA相比，PEAMA-b-聚乙二醇失活了α-淀粉酶和β-半乳糖苷酶，形成了可溶性的复合体。然后，通过添加阴离子聚丙烯酸(PAAc)，成功地将酶/PEAMA-b-PEG络合物回收出来。因此，聚乙二醇链段在PEAMA-b-PEG中的分散显然对调节这些酶的活性起着至关重要的作用，这表明聚乙二醇化的带电聚合物是一种新的CPPS候选材料，适用于大型和不稳定的酶。
The development of technology for on/off switching of enzyme activity is expected to expand the applications of enzyme in a wide range of research fields. We have previously developed a complementary polymer pair system (CPPS) that enables the activity of several enzymes to be controlled by a pair of oppositely charged polymers. However, it failed to control the activity of large and unstable a-amylase because the aggregation of the complex between anionic a-amylase and cationic poly(allylamine) (PAA) induced irreversible denaturation of the enzyme. To address this issue, we herein designed and synthesized a cationic copolymer with a poly(ethylene glycol) backbone, poly(N,N-diethylaminoethyl methacrylate)-block-poly(ethylene glycol) (PEAMA-b-PEG). In contrast to PAA, alpha-amylase and beta-galactosidase were inactivated by PEAMA-b-PEG with the formation of soluble complexes. The enzyme/PEAMA-b-PEG complexes were then successfully recovered from the complex by the addition of anionic poly(acrylic acid) (PAAc). Thus, dispersion of the complex by PEG segment in PEAMA-b-PEG clearly plays a crucial role for regulating the activities of these enzymes, suggesting that PEGylated charged polymer is a new candidate for CPPS for large and unstable enzymes.