Preparation of cross-linked enzyme aggregates of nitrile hydratase ES-NHT-118 from E.coli by macromolecular cross-linking agent  

作　　者：Liya Zhou Haixia Mou Jing Gao Li Ma Ying He Yanjun Jiang 

机构地区：[1]School of Chemical Engineering and Technology,Hebei University of Technology,Tianjin 300130,China [2]Hebei Provincial Key Lab of Green Chemical Technology and High Efficient Energy Saving,Hebei University of Technology,Tianjin 300130,China

出　　处：《Chinese Journal of Chemical Engineering》2017年第4期487-492,共6页中国化学工程学报（英文版）

基　　金：Supported by the National Nature Science Foundation of China(Nos.21306039,21276060,21276062);the Natural Science Foundation of Hebei Province(B2015202082,B2016202027);the Tianjin City High School Science&Technology Fund Planning Project(20140513)

摘　　要：Cross-linked enzyme aggregates(CLEAs) of nitrile hydratase(NHase) ES-NHT-118 from Escherichia coli were prepared by using ammonium sulfate as precipitating agent followed by cross-linking with dextran polyaldehyde for the first time. In this process, egg white was added as protein feeder for facilitating the formation of CLEAs. The optimal conditions of the immobilization process were determined. Michaelis constants(Km) of free NHase and NHase CLEAs were also determined. The NHase CLEAs exhibited increased stability at varied pH and temperature conditions compared to its free counterpart. When exposed to high concentrations of acrylamide, NHase CLEAs also exhibited effective catalytic activity.Cross-linked enzyme aggregates（CLEAs） of nitrile hydratase（NHase） ES-NHT-118 from Escherichia coli were prepared by using ammonium sulfate as precipitating agent followed by cross-linking with dextran polyaldehyde for the first time. In this process, egg white was added as protein feeder for facilitating the formation of CLEAs. The optimal conditions of the immobilization process were determined. Michaelis constants（Km） of free NHase and NHase CLEAs were also determined. The NHase CLEAs exhibited increased stability at varied pH and temperature conditions compared to its free counterpart. When exposed to high concentrations of acrylamide, NHase CLEAs also exhibited effective catalytic activity.

关 键 词：nitrile linking acrylamide linker ammonium facilitating counterpart Michaelis dextran constants 

分 类 号：Q814[生物学—生物工程]