Abstract
Dextranase is a unique biocatalyst that has high specificity and stereo-selectivity towards a complex biopolymer known as dextran. Dextranase has wide industrial application, but most of the time harsh environmental conditions adversely affect the functionality and stability of the enzyme. To overcome this issue, a covalent cross-linking immobilization method was adapted in the current study utilizing a nontoxic and biocompatible matrix known as chitosan. Chitosan hydrogel microspheres were synthesized using chitosan which exhibited noteworthy physical and mechanical strength. After treatment with glutaraldehyde, chitosan hydrogel microspheres were used for immobilization of dextranase. The kinetic characteristics of immobilized dextranase were compared with that of the soluble enzyme. A shift in optimum pH and temperature from 7.0 to 7.5 and 50 to 60 °C was observed after immobilization, respectively. Recycling efficiency, thermal stability, and activation energy distinctly improved after immobilization, whereas anchoring of substrate at the active site of the soluble dextranase exhibited an increase in K m with no change in V max after crosslinking. This technique involves the reduction in the size of carrier molecules (microspheres) that provide a larger surface area for improved immobilization efficiency. Therefore, it is concluded that increased stability and reusability of this immobilized biocatalyst makes it a promising aspirant for the utilization at commercial level.
Similar content being viewed by others
References
Dutta PK, Dutta J, Tripathi VS (2004) Chitin and chitosan: chemistry, properties and applications. J Sci Ind Res 63:20–31
Barbosa O, Torres R, Ortiz C, Berenguer-Murcia A, Rodrigues RC, Fernandez-Lafuente R (2013) Heterofunctional supports in enzyme immobilization: from traditional immobilization protocols to opportunities in tuning enzyme properties. Biomacromolecules 14:2433–2462
Basak S, Punetha VD, Bisht G, Bisht SS, Sahoo NG, Cho JW (2015) Recent trends of polymer-protein conjugate application in biocatalysis: a review. Polym Rev 55:163–198
Bezerra CS, de Farias Lemos CMG, de Sousa M, Gonçalves LRB (2015) Enzyme immobilization onto renewable polymeric matrixes: past, present, and future trends. J Appl Polym Sci 132:1–15
Zhao H, Cui Q, Shah V, Xu J, Wang T (2016) Enhancement of glucose isomerase activity by immobilizing on silica/chitosan hybrid microspheres. J Mol Catal B Enzym 126:18–23
Cantone S, Ferrario V, Corici L, Ebert C, Fattor D, Spizzo P, Gardossi L (2013) Efficient immobilisation of industrial biocatalysts: criteria and constraints for the selection of organic polymeric carriers and immobilisation methods. Chem Soc Rev 42:6262–6276
Datta S, Christena LR, Rajaram YRS (2013) Enzyme immobilization: an overview on techniques and support materials. 3 Biotech 3:1–9
Fang S, Chang J, Lee YS, Hwang EJ, Heo JB, Choi YL (2016) Immobilization of α-amylase from Exiguobacterium sp. DAU5 on chitosan and chitosan-carbon bead: its properties. J Appl Biol Chem 59:75–81
Hou C, Wang Y, Zhu H, Wei H (2016) Construction of enzyme immobilization system through metal-polyphenol assisted Fe3O4/chitosan hybrid microcapsules. Chem Eng J 283:397–403
Kumar-Krishnan S, Chakaravarthy S, Hernandez-Rangel A, Prokhorov E, Luna-Bárcenas G, Esparza R, Meyyappan M (2016) Chitosan supported silver nanowires as a platform for direct electrochemistry and highly sensitive electrochemical glucose biosensing. RSC Adv 6:20102–20108
Shahidi F, Arachchi JKV, Jeon YJ (1999) Food applications of chitin and chitosans. Trends Food Sci Technol 10:37–51
Hudson SM, Smith C (1998) Polysaccharides: chitin and chitosan: chemistry and technology of their use as structural materials. In: Kaplan DL (ed) Biopolymers from renewable resources. Springer, Berlin, pp 96–118
Paul W, Sharma CP (2000) Chitosan, a drug carrier for the 21st century: a review. STP Pharma Sci 10:5–22
Biró E, Németh ÁS, Sisak C, Feczkó T, Gyenis J (2008) Preparation of chitosan particles suitable for enzyme immobilization. J Biochem Biophys Methods 70:1240–1246
Guo Z, Bai S, Sun Y (2003) Preparation and characterization of immobilized lipase on magnetic hydrophobic microspheres. Enzyme Microb Technol 32:776–782
Budriene S, Gorochovceva N, Romaskevic T, Yugova L, Miezeliene A, Dienys G, Zubriene A (2005) β-Galactosidase from Penicillium canescens. Properties and immobilization. Open Chem 3:95–105
Park T-S, Jeong HJ, Ko J-A, Ryu YB, Park S-J, Kim D, Kim Y-M, Lee WS (2012) Biochemical characterization of thermophilic dextranase from a thermophilic bacterium, Thermoanaerobacter pseudethanolicus. J Microbiol Biotechnol 22:637–641
Khalikova E, Susi P, Korpela T (2005) Microbial dextran-hydrolyzing enzymes: fundamentals and applications. Microbiol Mol Biol Rev 69:306–325
Patel S, Goyal A (2011) Functional oligosaccharides: production, properties and applications. World J Microbiol Biotechnol 27:1119–1128
Ngah WW, Fatinathan S (2008) Adsorption of Cu (II) ions in aqueous solution using chitosan beads, chitosan–GLA beads and chitosan–alginate beads. Chem Eng J 143:62–72
Won K, Kim S, Kim KJ, Park HW, Moon SJ (2005) Optimization of lipase entrapment in Ca-alginate gel beads. Process Biochem 40:2149–2154
Nelson N (1944) A photometric adaptation of the Somogyi method for the determination of glucose. J Biol Chem 153:375–380
Somogyi M (1937) A reagent for the copper-iodometric determination of very small amounts of sugar. J Biol Chem 117:771–776
Lowry OH, Rosembrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Monsan P (1978) Optimization of glutaraldehyde activation of a support for enzyme immobilization. J Mol Catal 3:371–384
Sahiner N, Sagbas S, Aktas N (2016) Preparation of macro-, micro-, and nano-sized poly (Tannic acid) particles with controllable degradability and multiple biomedical uses. Polym Degrad Stabil 129:96–105
Chen H, Zhang Q, Dang Y, Shu G (2013) The effect of glutaraldehyde cross-linking on the enzyme activity of immobilized and beta-galactosidase on chitosan bead. Adv J Food Sci Technol 5:932–935
Li J, Du Y, Sun L, Liang H, Feng T, Wei YA, Yao P (2006) Chitosaneous hydrogel beads for immobilizing neutral protease for application in the preparation of low molecular weight chitosan and chito-oligomers. J Appl Polym Sci 101:3743–3750
Amaya-Delgado L, Hidalgo-Lara ME, Montes-Horcasitas MC (2006) Hydrolysis of sucrose by invertase immobilized on nylon-6 microbeads. Food Chem 99:299–304
Wang Y, Li Y, Liu S, Li B (2015) Fabrication of chitin microspheres and their multipurpose application as catalyst support and adsorbent. Carbohyd Polym 120:53–59
Giacomini C, Villarino A, Franco-Fraguas L, Batista-Viera F (1998) Immobilization of β-galactosidase from Kluyveromyces lactis on silica and agarose: comparison of different methods. J Mol Catal B Enzym 4:313–327
Zhang DH, Peng LJ, Wang Y, Li YQ (2015) Lipase immobilization on epoxy-activated poly (vinyl acetate-acrylamide) microspheres. Colloids Surf B Biointerfaces 129:206–210
Penzol G, Armisén P, Fernández-Lafuente R, Rodés L, Guisán JM (1998) Use of dextrans as long and hydrophilic spacer arms to improve the performance of immobilized proteins acting on macromolecules. Biotechnol Bioeng 60:518–523
Tischer W, Kasche V (1999) Immobilized enzymes: crystals or carriers? Trends Biotechnol 17:326–335
Rezakhani N, Parivar K, Khayati M, Etemadzade S (2015) Immobilization of protease in biopolymers (mixture of alginate-chitosan). J Paramed Sci 5:108–113
Abdel-Naby MA, Ismail AMS, Abdel-Fattah AM, Abdel-Fattah AF (1999) Preparation and some properties of immobilized Penicillium funiculosum 258 dextranase. Process Biochem 34:391–398
Altun S, Çakıroğlu B, Özacar M, Özacar M (2015) A facile and effective immobilization of glucose oxidase on tannic acid modified CoFe2O4 magnetic nanoparticles. Colloids Surf B Biointerfaces 136:963–970
El-Tanash AB, El-Baz E, Sherief AA (2011) Properties of Aspergillus subolivaceus free and immobilized dextranase. Eur Food Res Technol 233:735–742
Bayramoglu G, Arica MY, Genc A, Ozalp VC, Ince A, Bicak N (2016) A facile and efficient method of enzyme immobilization on silica particles via Michael acceptor film coatings: immobilized catalase in a plug flow reactor. Bioprocess Biosyst Eng 39:1–11
Tanxin Du, Bailing L, Xiaohui H, Baotan Z, Cuiming D (2009) Covalent immobilization of glucose oxidase onto Poly(St-GMA-NaSS) monodisperse microspheres via BSA as spacer arm. Appl Surf Sci 255:7937–7941
Acknowledgements
This research was funded and supported by The Karachi Institute of Biotechnology and Genetic Engineering (KIBGE), University of Karachi, Pakistan.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no conflict of interest exists.
Rights and permissions
About this article
Cite this article
Shahid, F., Aman, A., Nawaz, M.A. et al. Chitosan hydrogel microspheres: an effective covalent matrix for crosslinking of soluble dextranase to increase stability and recycling efficiency. Bioprocess Biosyst Eng 40, 451–461 (2017). https://doi.org/10.1007/s00449-016-1713-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00449-016-1713-7