Covalent attachment of lysozyme to cotton/cellulose materials: protein verses solid support activation

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-ptoluene 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.

Immobilization of lysozyme-cellulose amide-linked conjugates on cellulose I and II cotton nanocrystalline preparations

“…Previously the attachment of lysozyme to cotton (Edwards et al 2000) and wool (Wang et al 2009;Hu et al 2008) fabrics has been presented as a model for developing antimicrobial textile materials with selective activity. Cotton is absorbent, hypoallergenic, and eco-friendly, and enzyme-linked cotton has potential uses as a selective biomedical or hygienic material (Edwards et al 2011b). This paper examines nanocrystalline cotton conjugates of lysozyme with two distinct cellulose crystal structures (cellulose I and cellulose II) and the observed antimicrobial activity.…”

Nanocellulose conjugated with retinoic acid: its capability to adsorb aflatoxin B1

“…For its modification, 10 ml of nanocellulose at a concentration of 500 lg/ml was added to 5 ml of 5 % APTES (v/v), incubated for 30 min at 100°C (Edwards et al 2011). Then, APTES-modified nanocellulose was centrifuged at 5,000 rpm for 5 min and Cellulose washed with DW.…”

Protein crystal based materials for nanoscale applications in medicine and biotechnology

“…Unlike aldehydes, carbodiimides are “zero‐length” cross‐linkers, meaning they do not add additional atoms between the two conjugated molecules. This process has been utilized to produce stable collagen matrices (Park, Lee, Lee, & Suh, ), and protein‐based nanoparticles (Niknejad & Mahmoudzadeh, ), crystals (Hartje et al, ), and macrocomplexes (Lepvrier, Doigneaux, Moullintraffort, Nazabal, & Garnier, ), as well as to conjugate enzymes to cellulose‐based materials (Edwards, Prevost, Condon, Sethumadhavan, et al, ; Edwards, Prevost, Condon, & French, ; Edwards, Prevost, Condon, French, & Wu, ). 1‐Ethyl‐3‐(3‐dimethylaminopropyl)‐carbodiimide (EDC) is the most commonly used carbodiimide agent for protein conjugation, primarily due to its solubility in aqueous solutions.…”