Research review paper Recombinant CBM-fusion technology — Applications overview Carla Oliveira, Vera Carvalho, Lucília Domingues, Francisco M. Gama ⁎ CEB — Centre of Biological Engineering, University of Minho, 4710-057 Braga, Portugal abstract article info Article history: Received 12 September 2014 Received in revised form 6 February 2015 Accepted 9 February 2015 Available online 14 February 2015 Keywords: Carbohydrate-binding modules Heterologous expression systems Recombinant CBM-fusions Carbohydrate-binding activity Cellulose CBM applications Carbohydrate-binding modules (CBMs) are small components of several enzymes, which present an indepen- dent 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 diver- sity of CBMs, together with their unique properties, has long raised their attention for many biotechnological ap- plications. 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 ad- vances. Emphasis is given particularly to the applications of recombinant CBM-fusions in: (a) modification of fi- bers, (b) production, purification and immobilization of recombinant proteins, (c) functionalization of biomaterials and (d) development of microarrays and probes. © 2015 Elsevier Inc. All rights reserved. Contents 1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 358 1.1. CBM nomenclature . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 359 1.2. CBM classification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 359 1.3. Fold families . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 359 1.4. Types of CBMs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 359 1.5. Role of CBMs in CAZymes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 359 1.6. General applications of CBMs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 360 2. Applications of recombinant CBMs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 361 2.1. Modification of fibers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 361 2.2. Recombinant protein production and purification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 363 2.2.1. Affinity purification tools . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 363 2.2.2. Production of peptides and enzymes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 364 2.3. Immobilization of recombinant proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 364 Functionalization of biomaterials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 365 2.4. Microarrays and probes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 366 3. Perspectives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 366 Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 367 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 367 1. Introduction The molecular recognition of carbohydrates by proteins, namely gly- coside hydrolases, is essential in several biological processes, including cell–cell recognition, cellular adhesion, and host-pathogen interactions. Therefore, understanding the structural basis of the ligand specificity of Biotechnology Advances 33 (2015) 358–369 ⁎ Corresponding author at: CEB — Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar, 4710-057 Braga, Portugal. Tel.: +351 253 604 400; fax: +351 253 604 429. E-mail addresses: carlaoliveira@deb.uminho.pt (C. Oliveira), veracarvalho@deb.uminho.pt (V. Carvalho), luciliad@deb.uminho.pt (L. Domingues), fmgama@deb.uminho.pt (F.M. Gama). http://dx.doi.org/10.1016/j.biotechadv.2015.02.006 0734-9750/© 2015 Elsevier Inc. All rights reserved. Contents lists available at ScienceDirect Biotechnology Advances journal homepage: www.elsevier.com/locate/biotechadv