Characterization of supports activated with divinyl sulfone as a tool to immobilize and stabilize enzymes via multipoint covalent attachment. Application to chymotrypsin

Abstract

DVS supports are very suitable to stabilize enzymes via multipoint covalent attachment.

Tuning the catalytic properties of lipases immobilized on divinylsulfone activated agarose by altering its nanoenvironment

“…DVS-agarose beads support was prepared as previously described [52]. A volume of 1.5 mL DVS was stirred in 40 mL of 333 mM sodium carbonate at pH 12.5, giving a concentration of 0.35 M DVS, until the medium becomes homogeneous, then 2 g of agarose beads was added and leave under gentle agitation for 35 minutes.…”

“…This support can react with different moieties of the proteins, and produce some rigidification, and as a final enzyme-support reaction end point, the blocking of the support may be performed with different amino or thiol compounds [52]. Thus, we should have lipase molecules with identical orientation and degree of enzyme support multipoint covalent attachment.…”

Asymmetric Reduction of Prochiral Ketones by Using Self‐Sufficient Heterogeneous Biocatalysts Based on NADPH‐Dependent Ketoreductases

“…[31][32][33][34] We followed an empirical approach, until the best surfacea nd the best immobilization chemistries were established,t om aximize the enzyme loading (mg g À1 carrier )a nd the volumetric activity (U g À1 carrier )f or the asymmetric reduction of aryl ketone 1c as am odel substrate by using IPAa sasacrificial substrate for in situ recycling of NADPH (Table1). Nevertheless, we have succeeded in immobilizingi tb ys creening differentc hemistries and carriers, which resulted in an optimal immobilization protocol for this enzyme.…”

“…[31][32][33][34] We followed an empirical approach, until the best surfacea nd the best immobilization chemistries were established,t om aximize the enzyme loading (mg g À1 carrier )a nd the volumetric activity (U g À1 carrier )f or the asymmetric reduction of aryl ketone 1c as am odel substrate by using IPAa sasacrificial substrate for in situ recycling of NADPH (Table1). [34] Unfortunately,b yu sing Ag-DVS, the immobilization yield was lower than 40 %, and we did not observe the expected reaction product (Table1). [35] The immobilization yield was 100 %, but the immobilized enzyme lost all its initial activity.…”

Methacrylated Chitosan Based UV Curable Support for Enzyme Immobilization

“…An ideal support require some properties like high mechanical stability, hydrophobicity, biocompatibility, inertness toward enzymes, resistance to bacterial attacks and availability at low cost are the important 9,10 . There are three main categories of support material as lipophilic synthetic organic polymers such as polystyrene and polyacrylamide, hydrophilic biopolymer based on natural polysaccharides such as cellulose and dextran, and an inorganic solid such as glass and iron oxide 7,[11][12][13][14] . In spite of the many advantages of inorganic carriers (e.g., high stability against physical, chemical, and microbial degradation), most of the applications are performed with organic matrices.…”