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[来自胡萝卜软腐欧文氏菌的聚乙二醇化重组L-天冬酰胺酶:生产、性质及潜在应用]

[PEGylated recombinant L-asparaginase from Erwinia carotovora: Production, properties, and potential applications].

作者信息

Melik-Nubarov N S, Grozdova I D, Lomakina G Yu, Pokrovskaya M V, Pokrovski V S, Aleksandrova S S, Abakumova O Yu, Podobed O V, Grishin D V, Sokolov N N

出版信息

Prikl Biokhim Mikrobiol. 2017 Mar-Apr;53(2):164-72.

Abstract

N-hydroxysuccinimide ester of monomethoxy polyethylene glycol hemisuccinate was synthesized. It acylated amino groups in a molecule of recombinant L-asparaginase from Erwinia carotovora. A method of L-asparaginase modification by the obtained activated polyethylene glycol derivative was developed. The best results were produced by modification of the enzyme with a 25-fold excess of reagent relative to the enzyme tetramer. The modified L-asparaginase was isolated from the reaction mixture by gel filtration on Sepharose CL-6B. The purified bioconjugate did not contain PEG unbound to the protein, demonstrated high catalytic activity, and exhibited antiproliferative action on cell cultures.

摘要

合成了单甲氧基聚乙二醇半琥珀酸酯的N-羟基琥珀酰亚胺酯。它使来自胡萝卜软腐欧文氏菌的重组L-天冬酰胺酶分子中的氨基发生酰化。开发了一种用所得活性聚乙二醇衍生物修饰L-天冬酰胺酶的方法。相对于酶四聚体,用25倍过量的试剂修饰酶可产生最佳结果。通过在Sepharose CL-6B上进行凝胶过滤从反应混合物中分离出修饰的L-天冬酰胺酶。纯化的生物共轭物不含未与蛋白质结合的聚乙二醇,具有高催化活性,并对细胞培养物表现出抗增殖作用。

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