Technical Abstract: Enzyme-active cotton is a functional biocompatible material, and has potential applications as a sustainable material. With this in mind we have explored development of enzyme-active cotton with product potential as a disposable or reusable textile material. Lysozyme, which historically has been a well studied enzyme, was employed as a model enzyme since it has broad antimicrobial activity with antibacterial, antiviral, and antifungal applications. Lysozyme was covalently attached to three cotton fabrics using both an aqueous and organic based linking strategy. The antibacterial activity and several fiber properties that are signature characteristics of the enzyme-cellulose conjugate including zeta potential, and infra-red spectra were elucidated. Organic solvent-based carbodiimide coupling gave higher incorporation when compared with an aqueous based approach. The organic solvent approach with cotton twill using a carbodiimide-mediated coupling reaction gave the highest incorporation of lysozyme (16.1milligrams/gram cotton). Cotton spunlaced nonwoven gave slightly lower incorporation of lysozyme (12.8 milligrams/gram cotton) with highest antimicrobial activity, and woven print cloth yielded less (4 milligrams/gram cotton). The effect of storage on activity was monitored over a five month period, and as much as 60-70 per cent of enzyme activity remained after five months of storage.