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
- 171 Downloads
The carbohydrate-binding modules (CBMs) have emerged as an interesting alternative to enzymes for fibers modification, e.g. of pulp and paper. Glycosylation in CBMs is thought to have a key role in the improvement of cellulose fibers. Thus, in this work the non-glycosylated (CBM3mt) and glycosylated (CBM3wt) recombinant versions of CBM3 from Clostridium thermocellum CipA—both produced in Pichia pastoris—were studied. Binding assays showed that CBM3mt had a higher affinity for microcrystalline cellulose (Avicel) than CBM3wt. In addition, CBM3mt produced a much higher hydrophobization of Whatman paper than CBM3wt. However, the effects of the two CBM3s on pulp and paper were identical. The CBM3s did not affect the drainability of Eucalyptus globulus or a mixture of E. globulus and Pinus sylvestris pulps. On the other hand, both improved significantly strength-related properties of E. globulus papersheets, namely burst (up to 12 %) and tensile strength (up to 10 %) indexes. This is the first report showing the capacity of CBM3 from C. termocellum CipA to modify paper properties. The results showed that glycosylation did not influence the drainage of CBM3-treated pulps nor the properties of the produced papers. Thus, glycans in glycosylated CBM3 may not be related with fiber improvement, namely superior pulp drainage.
KeywordsRecombinant carbohydrate-binding module CBM3 Pichia pastoris expression system Glycosylation Cellulose Paper strength improvement
C. Oliveira acknowledges support from Fundação para a Ciência e a Tecnologia (FCT), Portugal (Grant SFRH/BDP/63831/2009). The authors thank the FCT GlycoCBMs Project REF. PTDC/AGR-FOR/3090/2012—FCOMP-01-0124-FEDER-027948, the FCT Strategic Project PEst-OE/EQB/LA0023/2013, and the Project “BioInd—Biotechnology and Bioengineering for improved Industrial and Agro-Food processes”, REF. NORTE-07-0124-FEDER-000028 Co-funded by the Programa Operacional Regional do Norte (ON.2—O Novo Norte), QREN, FEDER.
Conflict of interest
The authors declare that they have no conflict of interest.
- Cadena EM, Chriac AI, Pastor FIJ, Diaz P, Vidal T, Torres AL (2010) Use of cellulases and recombinant cellulose binding domains for refining TCF kraft pulp. Biotechnol Prog 26:960–967Google Scholar
- Kim DW, Jang YH, Kim CS, Lee NS (2001) Effect of metal ions on the degradation and adsorption of two cellobiohydrolases on microcrystalline cellulose. Bull Korean Chem Soc 22:716–720Google Scholar
- Oliveira C, Carvalho V, Domingues L, Gama FM (2015) Recombinant CBM-fusion technology: applications overview. Biotechnol Adv 33:358–369Google Scholar
- Pinto R (2006) Production of cellulose-binding domains by proteolysis; studies on the adsorption and modification of cellulose fibres. Ph.D. thesis, University of Minho, BragaGoogle Scholar
- Pinto R, Amaral E, Costa AP, Gama FM, Duarte AP (2004b) Improving papermaking with cellulose-binding domains. CIADICYP 2004: congresso Iberoamericano de Investigacion en Cellulose y Papel, vol. 2004. Córdoba, Spain, pp 303–305. ISBN 84-7498-504-8Google Scholar
- Shi XR, Zheng F, Pan RH, Wang J, Ding SJ (2014) Engineering and comparative characteristics of double carbohydrate binding modules as a strength additive for papermaking applications. Bioresources 9:3117–3131Google Scholar
- Tormo J, Lamed R, Chirino AJ, Morag E, Bayer EA, Shoham Y et al (1996) Crystal structure of a bacterial family-III cellulose-binding domain: a general mechanism for attachment to cellulose. EMBO J 15:5739–5751Google Scholar
- Yokota S, Matsuo K, Kitaoka T, Wariishi H (2009) Retention and paper-strength characteristics of anionic polyacrylamides conjugated with carbohydrate-binding modules. Bioresources 4:234–244Google Scholar