Cellulase immobilization on superparamagnetic nanoparticles for reuse in cellulosic biomass conversion

Qing Song; Yu Mao; Mark Wilkins; Fernando Segato; Rolf Prade; Qing Song; Yu Mao; Mark Wilkins; Fernando Segato; Rolf Prade

doi:10.3934/bioeng.2016.3.264

AIMS Bioengineering

2016, Volume 3, Issue 3: 264-276. doi: 10.3934/bioeng.2016.3.264

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Research article

Cellulase immobilization on superparamagnetic nanoparticles for reuse in cellulosic biomass conversion

1.
Department of Immunology, Institute of Tuberculosis Control, Institute of Human Virology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, China; Key Laboratory of Tropical Diseases Control (Sun Yat-sen University), Ministry of Education, Guangzhou 510080, China
2.
Department of Biosystems Engineering, Oklahoma State University, Stillwater, OK 74078, USA
3.
Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK 74078, USA

Received: 31 May 2016 Accepted: 14 July 2016 Published: 17 July 2016

Current cellulosic biomass hydrolysis is based on the one-time use of cellulases. Cellulases immobilized on magnetic nanocarriers offer the advantages of magnetic separation and repeated use for continuous hydrolysis. Most immobilization methods focus on only one type of cellulase. Here, we report co-immobilization of two types of cellulases, β-glucosidase A (BglA) and cellobiohydrolase D (CelD), on sub-20 nm superparamagnetic nanoparticles. The nanoparticles demonstrated 100% immobilization efficiency for both BglA and CelD. The total enzyme activities of immobilized BglA and CelD were up to 67.1% and 41.5% of that of the free cellulases, respectively. The immobilized BglA and CelD each retained about 85% and 43% of the initial immobilized enzyme activities after being recycled 3 and 10 times, respectively. The effects of pH and temperature on the immobilized cellulases were also investigated. Co-immobilization of BglA and CelD on MNPs is a promising strategy to promote synergistic action of cellulases while lowering enzyme consumption.
- enzyme co-immobilization,
- β-glucosidase,
- cellobiohydrolase,
- superparamagnetic nanoparticles,
- cellulase reuse
Citation: Qing Song, Yu Mao, Mark Wilkins, Fernando Segato, Rolf Prade. Cellulase immobilization on superparamagnetic nanoparticles for reuse in cellulosic biomass conversion[J]. AIMS Bioengineering, 2016, 3(3): 264-276. doi: 10.3934/bioeng.2016.3.264

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Abstract

Current cellulosic biomass hydrolysis is based on the one-time use of cellulases. Cellulases immobilized on magnetic nanocarriers offer the advantages of magnetic separation and repeated use for continuous hydrolysis. Most immobilization methods focus on only one type of cellulase. Here, we report co-immobilization of two types of cellulases, β-glucosidase A (BglA) and cellobiohydrolase D (CelD), on sub-20 nm superparamagnetic nanoparticles. The nanoparticles demonstrated 100% immobilization efficiency for both BglA and CelD. The total enzyme activities of immobilized BglA and CelD were up to 67.1% and 41.5% of that of the free cellulases, respectively. The immobilized BglA and CelD each retained about 85% and 43% of the initial immobilized enzyme activities after being recycled 3 and 10 times, respectively. The effects of pH and temperature on the immobilized cellulases were also investigated. Co-immobilization of BglA and CelD on MNPs is a promising strategy to promote synergistic action of cellulases while lowering enzyme consumption.

References

[1]	Menon V, Rao M (2012) Trends in bioconversion of lignocellulose: Biofuels, platform chemicals &biorefinery concept. Prog Energy Combust Sci 38: 522–550. doi: 10.1016/j.pecs.2012.02.002
[2]	Wang H, Gurau G, Rogers RD (2012) Ionic liquid processing of cellulose. Chem Soc Rev 41: 1519–1537. doi: 10.1039/c2cs15311d
[3]	Bhat M, Bhat S (1997) Cellulose degrading enzymes and their potential industrial applications. Biotechnol Adv 15: 583–620. doi: 10.1016/S0734-9750(97)00006-2
[4]	Lynd LR, Weimer PJ, van Zyl WH, et al. (2002) Microbial cellulose utilization: fundamentals and biotechnology. Microbiol Mol Biol Rev 66: 506–577. doi: 10.1128/MMBR.66.3.506-577.2002
[5]	Lee SM, Jin LH, Kim JH, et al. (2010) β-glucosidase coating on polymer nanofibers for improved cellulosic ethanol production. Bioprocess Biosyst Eng 33: 141–147. doi: 10.1007/s00449-009-0386-x
[6]	Sheldon RA (2007) Enzyme immobilization: the quest for optimum performance. Adv Synth Catal 349: 1289–1307. doi: 10.1002/adsc.200700082
[7]	Mateo C, Palomo JM, Fernandez-Lorente G, et al. (2007) Improvement of enzyme activity, stability and selectivity via immobilization techniques. Enzyme Microb Technol 40: 1451–1463. doi: 10.1016/j.enzmictec.2007.01.018
[8]	Mohamad NR, Marzuki NHC, Buang NA, et al. (2015) An overview of technologies for immobilization of enzymes and surface analysis techniques for immobilized enzymes. Biotechnol Biotechnol Equip 29: 205–220. doi: 10.1080/13102818.2015.1008192
[9]	Chim-Anage P, Kashiwagi Y, Magae Y, et al. (1986) Properties of cellulase immobilized on agarose gel with spacer. Biotechnol Bioeng 28: 1876–1878. doi: 10.1002/bit.260281215
[10]	Isgrove F, Williams R, Niven G, et al. (2001) Enzyme immobilization on nylon-optimization and the steps used to prevent enzyme leakage from the support. Enzyme Microb Technol 28: 225–232. doi: 10.1016/S0141-0229(00)00312-4
[11]	Li C, Yoshimoto M, Fukunaga K, et al. (2007) Characterization and immobilization of liposome-bound cellulase for hydrolysis of insoluble cellulose. Bioresour Technol 98: 1366–1372. doi: 10.1016/j.biortech.2006.05.028
[12]	Georgelin T, Maurice V, Malezieux B, et al. (2010) Design of multifunctionalized γ-Fe₂O₃@SiO₂ core–shell nanoparticles for enzymes immobilization. J Nanopart Res 12: 675–680. doi: 10.1007/s11051-009-9757-0
[13]	Wang P, Hu X, Cook S, et al. (2009) Influence of silica-derived nano-supporters on cellobiase after immobilization. Appl Biochem Biotechnol 158: 88–96. doi: 10.1007/s12010-008-8321-1
[14]	Verma ML, Chaudhary R, Tsuzuki T, et al. (2013) Immobilization of β-glucosidase on a magnetic nanoparticle improves thermostability: Application in cellobiose hydrolysis. Bioresour Technol 135: 2–6. doi: 10.1016/j.biortech.2013.01.047
[15]	Wood TM, Bhat KM (1988) Methods for measuring cellulase activities. Methods Enzymol 160: 87–112. doi: 10.1016/0076-6879(88)60109-1
[16]	Saha BC, Bothast RJ (1996) Production, purification, and characterization of a highly glucose-tolerant novel beta-glucosidase from Candida peltata. Appl Environ Microbiol 62: 3165–3170.
[17]	Segato F, Damásio AR, Gonçalves TA, et al. (2012) High-yield secretion of multiple client proteins in Aspergillus. Enzyme Microb Technol 51: 100–106. doi: 10.1016/j.enzmictec.2012.04.008
[18]	Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254. doi: 10.1016/0003-2697(76)90527-3
[19]	Shapiro AL, Viñuela E, Maizel JV Jr (1967) Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochem Biophys Res Commun 28: 815–820. doi: 10.1016/0006-291X(67)90391-9
[20]	Caruntu D, Caruntu G, Chen Y, et al. (2004) Synthesis of variable-sized nanocrystals of Fe₃O₄ with high surface reactivity. Chem Mater 16: 5527–5534. doi: 10.1021/cm0487977
[21]	Lu Y, Yin Y, Mayers BT, et al. (2002) Modifying the surface properties of superparamagnetic iron oxide nanoparticles through a sol-gel approach. Nano Lett 2: 183–186. doi: 10.1021/nl015681q
[22]	Caruntu D, Caruntu G, O'Connor CJ (2007) Magnetic properties of variable-sized Fe₃O₄ nanoparticles synthesized from non-aqueous homogeneous solutions of polyols. J Phys D Appl Phys 40: 5801–5809.
[23]	Wahajuddin, Arora S (2012) Superparamagnetic iron oxide nanoparticles: magnetic nanoplatforms as drug carriers. Int J Nanomedicine 7: 3445–3471.
[24]	Kannan K, Jasra RV (2010) Improved catalytic hydrolysis of carboxy methyl cellulose using cellulase immobilized on functionalized meso cellular foam. J Porous Mat 18: 409–416.
[25]	Jung C (2000) Insight into protein structure and protein–ligand recognition by Fourier transform infrared spectroscopy. J Mol Recognit 13: 325–351.
[26]	Gotić M, Musić S (2007) Mössbauer, FT-IR and FE SEM investigation of iron oxides precipitated from FeSO₄ solutions. J Mol Struct 834: 445–453.
[27]	Gai L, Li Z, Hou Y, et al. (2010) Preparation of core-shell Fe₃O₄/SiO₂ microspheres as adsorbents for purification of DNA. J Phys D Appl Phys 43: 5625–5634.
[28]	Zhang D, Hegab HE, Lvov Y, et al. (2016) Immobilization of cellulase on a silica gel substrate modified using a 3-APTES self-assembled monolayer. Springerplus 5: 48. doi: 10.1186/s40064-016-1682-y
[29]	Zhou J, Zhang J, Gao W (2014) Enhanced and selective delivery of enzyme therapy to 9L-glioma tumor via magnetic targeting of PEG-modified, β-glucosidase-conjugated iron oxide nanoparticles. Int J Nanomedicine 9: 2905–2917.
[30]	Abraham RE, Verma ML, Barrow CJ, et al. (2014) Suitability of magnetic nanoparticle immobilised cellulases in enhancing enzymatic saccharification of pretreated hemp biomass. Biotechnol Biofuels 7: 90–101. doi: 10.1186/1754-6834-7-90
[31]	Ladeira SA, Cruz E, Delatorre AB, et al. (2015). Cellulase production by thermophilic Bacillus sp. SMIA-2 and its detergent compatibility. Electron J Biotechn 18: 110–115.

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