Abstract
Covalent attachment of enzymes to cellulosic materials like cotton is of interest where either release or loss of enzyme activity over time needs to be avoided. The covalent attachment of an enzyme to a cellulosic substrate requires either activation of a protein side chain or an organic functional group on the cellulosic substrate. Use of a water soluble carbodiimide to create an amide linkage as the covalent attachment between the enzyme and substrate represents an aqueous-based alternative which may be preferred for textile processes. Here we describe an amide bond-mediated lysozyme immobilization applied to cotton where either the carboxylate side chains of the protein or pendant carboxylates in a citrate, cross-linked cotton support are activated as the O-acyl-isourea intermediate, and the reactive amino nucleophiles are derived from amino-silanized cotton and the protein’s amino side chains, respectively. A comparison is made of the two activation approaches to covalently link lysozyme to two different cotton fabrics using the water soluble carbodiimide 1-cyclohexyl-3-(2-morpholinoethyl)-carbodiimide-metho-p-toluene sulfonate. A comparison of the resulting enzyme activities of lysozyme on two different cotton supports showed that linking lysozyme to citrate crosslinked cotton gave higher activity than on aminosilanized cotton. The lysozyme-cellulose conjugate formed on the citrate crosslinked nonwoven cotton fabric gave the highest yield and antimicrobial activity.
Similar content being viewed by others
References
Alonso D, Gimeno M, Olayo R, Vazquez-Torres H, Sepulveda-Sanchez J, Shirai K (2009) Cross-linking chitosan into UT-irradiated cellulose fibers for the preparation of antimicrobial-finished textiles. Carbohydr Polym 77:536–543
Antonio BF, Valle-Vigon P, Sevilla M (2010) Synthesis of colloidal nanoparticles of a tunable mesopore size and their application to the adsorption of biomolecules. J Colloid Interface Sci 349:173–180
Baltazar-y-Jimenez A, Bismarck A (2007) Wetting behavior, moisture up-take and electrokinetic properties of lignocellulosic fibres. Cellulose 14:115–127
Beddows CG (1988) The immobilization of enzymes onto hydrolyzed polyethylene-g-co-2-HEMA. J Appl Polym Sci 35:135–144
Bismarck A, Springer J, Mohanty AK, Hinrichsen G, Khan MA (2000) Characterization of several modified jute fibers using zeta-potential measurements. Colloid Polym Sci 278:229–235
Blake CCF, Mair GA, North ACT, Phillips DC, Sarma VR (1967a) On the conformation of hen egg-white lysozyme molecule. Proc Roy Soc Ser B 167:365–377
Blake CCF, Johnson LN, Mair GA, North ACT, Phillips DC, Sarma VR (1967b) Crystallographic studies of the activity of hen egg-white lysozyme. Proc Roy Soc Lond B 167:378–388
Branen JK, Davidson MP (2004) Enhancement of nisin, lysozyme, and monolaurin antimicrobial activities by ethylenediaminetetraacetic acid and latoferrin. Int J of Food Microbiol 90:63–74
Cisani G, Varaldo PE, Pompei R, Valiesena S, Satra G (1989) Cell fusion induced by herpes simplex is inhibited by hen egg-white lysozyme. Microbios 59:73–83
Ding H-M, Shao L, Liu R-J, Xiao Q-G, Chen J-F (2005) Silica nanotubes for lysozyme immobilization. J Colloid Interface Sci 290:102–106
Dragoni I, Balzaretti C, Rossini S, Rossi L, Dell’Orto V, Baldi A (2011) Detection of hen lysozyme on proteic profiles of Grana Padano cheese through SELDI-TOFI MS high-throughput technology during the ripening process. Food Anal Methods 4:233–339
Edwards JV, Sethumadhavan K, Ullah AHJ (2000) Conjugation and modeled structure/function analysis of lysozyme on glycine esterified cotton cellulose fibers. Bioconjugate Chem 11:469–473
Edwards JV, Eggleston G, Yager DR, Cohen IK, Diegelmann RF, Bopp AF (2002) Design preparation and assessment of citrate-linked monosaccharide cellulose conjugates with elastase-lowering activity. Carbohydr Polym 50:305–314
Edwards JV, Prevost N, Condon B, Sethumadhavan K, Ullah J (2011) Immobilization of lysozyme on cotton fabrics: synthesis, characterization and activity. AATCC 11:73–79
Gao P, Cai P (2008) The boundary molecules in a lysozyme pattern exhibit preferential antibody binding. Langmuir 24:10334–10339
Grimsley JK, Singh WP, Wild JR, Giletto A (2001) A novel, enzyme-based method for the wound—surface removal and decontamination of organophosphorus nerve agents. In: Edwards JV, Vigo TL (eds) Bioactive fibers and polymer, ACS symposium series 792. American Chemical Society, Washington
Gupta P, Bajpai M, Bajpai SK (2008) Textile technology: investigations of antibacterial properties of silver nanoparticle-loaded poly (acrylamide-co-itaconic acid)-grafted cotton fabric. J Cotton Sci 12:280–286
Haynes DH, Kowalsky A, Pressman BC (1969) Carboxyl group modification and the activity of lysozyme. J Biol Chem 244:505–508
Hirano S (1999) Chitin and chitosan as novel biotechnological materials. Polym Int 48:732–734
Ibrahim NA, Gouda M, El-Shafei AM, Abdel-Fatah OM (2007) Antimicrobial activity of cotton fabrics containing immobilized enzymes. J Appl Polym Sci 104:1754–1761
Janolino VG, Swaisgood HE (1982) Analysis and optimization of methods using water-soluble carbodiimide for immobilization of biochemicals to porous glass. Biotechnol Bioeng 24:1069–1080
Jeanloz RW, Sharon N, Flowers HM (1963) The chemical structure of a disaccharide isolated from Micrococcus lysodeikticus cell wall. Biochem Biophys Res Commun 13:20–25
Klibanov AM (1983) Immobilized enzymes and cells as practical catalysts. Science 219:722–727
Kotwal S, Shankar V (2009) Immobilized invertase. Biotechnol Adv 22:311–322
Kubiak-Ossowska K, Mulheran PA (2010) What governs protein adsorption and immobilization at a charged solid surface? Langmuir 26:7690–7694
Lampis G, Deidda D, Pinza M, Pompei R (2001) Enhancement of anti-herpetic activity of glycyrrhizic acid by physiological proteins. Antivir Chem Chemother 12:125–131
Lee-Huang S, Maiorov V, Huang PL, Ng A, Hee CL, Chang Y-T, Kallenbach N, Huang PL, Chen H-C (2005) Structural and functional modeling of human lysozyme reveals a unique nonapeptide, HL9, with anti-HIV activity. Biochemistry 44:4648–4655
Lou C-W (2008) Process technology and properties evaluation of a chitosan-coated tencel/cotton nonwoven fabric as a wound dressing. Fibers Polym 9:286–292
Madhumati R, Luckarift HR, Sarsenova A, Wild FR, Ramanculov EK, Olsen EV, Simonian A (2009) Lysozyme-mediated formation of protein-silica nano-composites for biosensing applications. Colloids Surf B Biointerfaces 73:58–64
Minier M, Salmain M, Yacoubi N, Barbes L, Methivier C, Zanna S, Pradier C-M (2005) Covalent immobilization of lysozyme on stainless steel interface spectroscopic characterization and measurement of enzymatic activity. Langmuir 21:5957–5965
Ouahidi I, Hamsas AY, Aarab L (2011) Modulation of egg white protein allerginicity under physical and chemical treatment. Food Agric Immunol 22:57–68
Parikh DV, Edwards JV, Condon BD, Parikh AD (2008) Silver-carboxylate ion-paired alginate and carboxymethylated cotton with antimicrobial activity. AATCC Review 8:38–43
Pechkova E, Tripathi SK, Nicolini C (2010) Comparison of lysozyme crystals grown by apa and classical hanging drop method. Image from the RCSB PDB (http://www.pdb.org) of PDB ID: 3IJU. doi:10.2210/pdb3iju/pdb
Pettersen EF, Goddard TD, Huang CC, Couch GS, Greenblatt DM, Meng EC, Ferrin TE (2004) UCSF Chimera–a visualization system for exploratory research and analysis. J Comput Chem 25:1605–1612
Philips DC (1966) The three-dimensional structure of an enzyme molecule. Sci Amer 215:78–90
Purwar R, Mishra P, Joshi M (2008) Antibacterial finishing of cotton textiles using neem extract. AATCC Review 8:36–43
Salton MRJ, Ghuysen JM (1959) The structure of di- and tetra-saccharides released from cell walls by lysozyme and Streptomyces F1 enzyme and the β (1-4)Ν-acetylhexosaminidase activity of these enzymes. Biochim Biophys Acta 36:552–554
Salton MRJ, Ghuysen JM (1960) Acetylhexosamine compounds enzymatically released from micrococcus lysodeikticus cell walls. Biochim Biophys Acta 45:355–363
Samaranayake YH, Smaranayake LP, Pow EHN, Beena VT, Yeung KWS (2001) Antifungal effects of lysozyme and lactoferrin against genetically similar, sequential candida albicans isolates from a human immunodeficiency virus-infected southern Chinese cochort. J Clin Microbiol 39:3296–3302
Thilagavathi G, Kannaian T (2008) Application of prickly chaff (Achyranthes aspera Linn.) leaves as herbal antimicrobial finish for cotton fabric used in healthcare textiles. Natural Product Radiance 7:330–334
Tomsic B, Simoncic B, Orle B, Zerjav M, Schroers H, Simoncic A, Samardziaz Z (2009) Antimicrobial activity of AgCl embedded in a silica matrix on cotton fabric. Carbohydr Polym 75:618–626
Wang Q, Fann X, Hu Y, Yuan J, Cui L, Wang P (2009) Antibacterial functionalization of wool fabric via immobilizing lysozymes. Bioprocess Biosyst Eng 32:633–639
Weber P, Kratzin H, Brockow K, Ring J, Steinhart H, Paschke A (2009) Lysozyme in wine: a risk evaluation for consumers allergic to hen’s egg. Mol Nutr Food Res 53:1469–1477
Wedmore I, McManus JG, Pusateri AE, Holcomb JB (2006) A special report on the chitosan-based hemostatic dressing experience in current combat operations. J Trauma 60:655–658
Xie T, Wang A, Huang L, Li H, Chen Z, Wang Q, Yin X (2009) Recent advance in the support and technology used in enzyme immobilization. Afr J Biotechnol 8:4724–4733
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Edwards, J.V., Prevost, N.T., Condon, B. et al. Covalent attachment of lysozyme to cotton/cellulose materials: protein verses solid support activation. Cellulose 18, 1239–1249 (2011). https://doi.org/10.1007/s10570-011-9563-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10570-011-9563-6