Recombinant CBM-fusion technology — Applications overview

Abstract

Carbohydrate-binding modules (CBMs) are small components of several enzymes, which present an independent fold and function, and specific carbohydrate-binding activity. Their major function is to bind the enzyme to the substrate enhancing its catalytic activity, especially in the case of insoluble substrates. The immense diversity of CBMs, together with their unique properties, has long raised their attention for many biotechnological applications. Recombinant DNA technology has been used for cloning and characterizing new CBMs. In addition, it has been employed to improve the purity and availability of many CBMs, but mainly, to construct bi-functional CBM-fused proteins for specific applications. This review presents a comprehensive summary of the uses of CBMs recombinantly produced from heterologous organisms, or by the original host, along with the latest advances. Emphasis is given particularly to the applications of recombinant CBM-fusions in: (a) modification of fibers, (b) production, purification and immobilization of recombinant proteins, (c) functionalization of biomaterials and (d) development of microarrays and probes.

Functional and structural characterization of a potent GH74 endo‐xyloglucanase from the soil saprophyte Cellvibrio japonicus unravels the first step of xyloglucan degradation

“…Indeed, a particular challenge in CBM characterization is the difficulty in obtaining soluble recombinant production in E. coli as a result of improper protein folding [29]. Recombinant CBM-fusion technology has been used extensively for various applications [30], and we were particularly inspired by the specific use of CBM-GFP fusion constructs to facilitate the production and purification of CBMs and CBM-tagged fusion proteins [31][32][33][34][35] and to make probes for the determination of surface accessibility of cellulose [36,37]. Hence, both CBMs were fused to an N-terminal superfolder GFP (sfGFP) domain (Fig.…”

Modification of paper properties using carbohydrate-binding module 3 from the Clostridium thermocellum CipA scaffolding protein produced in Pichia pastoris: elucidation of the glycosylation effect

“…Due to their independent fold and function, CBMs find application in different fields of biotechnology (Gomes et al 2015;Oliveira et al 2015). CBMs can be obtained from fungal and bacterial cellulases and hemicellulases through proteolysis (Lemos et al 2000;Pinto et al 2004a) or by recombinant DNA technology, mainly in fusion with a partner (reviewed in Oliveira et al 2015).…”

Bioprospecting thermostable cellulosomes for efficient biofuel production from lignocellulosic biomass

“…Apart from this, many researchers have also reported other applications of cellulosomal enzymes including removal of bacterial biofilms (Juturu and Wu 2014); enhancement of mechanical and physical properties of paper (Cadena et al 2010;Shi et al 2014); in textile, detergent and food processing (Juturu and Wu 2014;Karmakar and Ray 2011;Kuhad et al 2011;Oliveira et al 2015).…”