Enzymatic microreactors in chemical analysis and kinetic studies

Silica-immobilized enzyme reactors; application to cholinesterase-inhibition studies

“…This bioencapsulation strategy therefore provides an economical and rapid route for synthesizing IMER systems with a number of advantages including; minimal preparation time, high immobilization efficiency and excellent stability. IMERs could be designed to contain an enzyme for biocatalysis or organic synthesis, for rapid screening in medical diagnostics and therapy or for developing IMER columns for affinity chromatography [1,2,25]. In addition, coimmobilization of multienzyme systems is also possible.…”

“…Immobilized enzyme reactors (IMERs) have found application in catalysis and have also been used with a wide variety of receptor proteins for substrate interaction and inhibition studies [1][2][3][4][5][6][7]. The main advantages of immobilized enzyme systems are stability and reusability.…”

Silica-Immobilized Enzyme Reactors

“…The ability to study enzyme kinetic characteristics using IMER systems by calculation of Michaelis-Menten Kinetics has been demonstrated to confirm that the immobilized enzyme is not significantly hindered by its orientation and binding. [8,34] Lineweaver-Burk plots of reaction rate vs. substrate concentration are readily achievable in flowthrough IMER systems from which the Michaelis constant (K m ) and the maximum enzyme velocity (V max ) can be calculated and mechanisms of inhibition (competitive, noncompetitive etc.) can be determined.…”

“…The use of IMERs for protein digestion, however, has been reviewed extensively elsewhere and will not be covered herein. [6][7][8][9][10] A variety of matrices have been investigated over the years for enzyme immobilization or encapsulation, but for focus, the emphasis within this review is limited to IMERs that utilize silica as a scaffold. The use of silica-based IMERs has received increasing attention since their appearance in the early 1970's and since then, the use and diversity of applications has grown steadily ( Figure 1).…”

Imobilização de enzimas em suportes cromatográficos: uma ferramenta na busca por substâncias bioativas

“…2,19 A segunda, engloba os biorreatores desenvolvidos para triagem de substratos e para os estudos das características cinéticas das enzimas. 20 Nessa segunda classe, os IMER têm sido amplamente utilizados para separação e identificação de metabólitos, 21 nos estudos de metabolismo dos fármacos, 22,23 para análise e síntese enantiosseletiva, 2,24-26 na digestão proteolítica de proteínas on-line, 27 no controle biotecnoló-gico de fármacos, 28 e para a identificação de substratos e/ou inibidores, na busca por novos ligantes. [29][30][31][32][33][34][35] Também merece destaque o uso de enzimas imobilizadas como biossensores em aplicações biomédicas 36 ou como órgãos artificiais.…”