Calmodulin-mediated reversible immobilization of enzymes

A Chimeric Fusion Protein Engineered with Disparate Functionalities—Enzymatic Activity and Self–assembly

“…12 OPH activity is minimally affected in a CBD fusion, 14 but enzymatic activity is decreased by 1 order of magnitude in a calmodulin-OPH fusion. 16 Additionally, there is no measurable difference in HRP activity between the wild-type and a CBD-HRP fusion. 15 Cross-comparison of the different fusions does not provide specific insight into the different protein engineering problems, but does highlight the success of the HS-Adh-H fusion and leads to the simple observation that each case is unique.…”

“…Also described are OPH 14 and horseradish peroxide (HRP) 15 fusions to a cellulose binding domain (CBD), for immoblization on the cellulose surfaces and calmodulin fusions to OPH and β-lactamase for reversible immobilization on to appropriately modified surfaces. 16 Perhaps the most common use of protein fusions is in biotechnologies for heterologous expression of recombinant proteins and in the purification of such products. [17][18][19] In expression and purification technologies, the fusion is often temporary as cleavage of the fusion generally occurs en route to the final product.…”

Affinity Interactions as a Tool for Protein Immobilization

“…The reversibility of the methods, enabling the support reloading, and the possibility of direct enzyme immobilization from crude cell lysate without additional purification steps contribute to the better applicability of these methods in practice. Mild reaction conditions and relative simplicity of these immobilization processes should also be emphasized (Andreescu et al, 2006;Brena & Batista-Viera, 2006;Bucur et al, 2004;Clare et al, 2001;Costa et al, 2005;Daunert et al, 2007;Kumada et al, 2010;Saleemuddin, 1999).…”

“…Not negligible advantage of use of fusion protein approach consists in the possibility to attach one type of fusion partner to different proteins of interest. This fact enables the usage of one support type, preparation of which can sometime be very expensive or complicated, for immobilization of various enzymes (Arnau et al, 2006;Chern & Chao, 2005;Costa et al, 2005;Daunert et al, 2007;Nilsson et al, 1997;Sørensen & Mortensen, 2005;Rao et al, 1998;Saleemuddin, 1999;Terpe, 2003).…”

Biosensors for endocrine disruptors

“…OPH is an ideal biocatalyst because of its broad substrate specifi city, stability over broad pH and temperature ranges and lack of requirement for expensive cofactors [101]. There are numerous optical and electrochemical methods [78,[102][103][104][105] used for detecting and identifying OP agents.…”