Design And Application Of Novel Metal Organic Framewor Enzyme Immobilized Bioreactor

Immobilized enzyme preserve the specific activity of enzyme and allows the repeated use of enzyme.Metal organic frameworks(MOF)have high porosity and specific surface area,controllable pore chemical properties and have the potential of being the carrier of immobilized enzyme.But the pore size of MOFs is less than 1 nm generally,while the size of enzyme is range from 3-5 nm.In this work,we prepared the enzyme micro reactors through two different methods,including layer by layer self-assembly and supramolecular self-assembly,and realized the reuse of enzyme.The main work of this paper is as follows:(1)A novel metal organic framework-monolith hybrid micro reactor,which was prepared through the layer by layer self-assembly of metal organic frameworks,was used for trypsin immobilized.Firstly,the surface of monolith was pretreated with 1-(3-aminopropyl)imidazole.And zeolitic imidazolate framework(ZIF-8)on the monolith surface was grown by self-assembly using 2-methyl imidazole and Zn2+.The results of Infrared X spectroscopy,X-ray diffraction nd other results showed the successfully growth of ZIF-8(2)Using metal chelating and chemical coupling method to immobilized trypsin on the surface of metal organic framework-monolithic hybrid materials.To hydrolysis protein,the micro reaction can get ideal enzymatic hydrolysis result for only 43 s.RP-HPLC and mass spectrometry results showed that the hydrolysis polypeptide fragment sequence coverage of cytochrome C and bovine serum albumin were 100%and 63.6%.And after 10 cycles,the immobilized enzyme micro reactor which was prepared by chemical coupling of the can keep the Cytochrome C sequence coverage above 90%,proving that the immobilized enzyme has good stability and repeatability.In addition,we designed a nano multienzyme catalytic system through supramolecular self-assembly.Firstly,MIL-101 was synthesized and was amino modified by PEI50,PEI 100,cystamine and hexamethylenediamine,respectively.Then,two layers of HKUST-1 were synthesized on the surface of MIL-101 and the carbonic anhydrase,formate dehydrogenase and glutamate dehydrogenase was embedded into HKUST-1 in the same time.The C02 adsorbed by MIL-101 was released in the catalytic reaction,captured by formate dehydrogenase and transferred into carbonic acid.When four different modification systems were compared,the PEI 100 modified system reached the highest yield within 2 h and the yield was 23.86 ?mol.