Synergism between Cutinase and Pectinase in the Hydrolysis of Cotton Fibers’ Cuticle

Abstract

Scouring is one of the initial steps in the processing of natural textile fibers (e.g., cotton), performed to remove waxes and pectins, together with spinning oils and other impurities of the plant cell cuticle. Traditional chemical bleaching with boiling NaOH led to harsh removal of the entire fabric’s cuticle waxy layer accompanied by an unwanted alkaline waste. Extracellular lytic enzymes such as lipases, cellulases and pectinases play an essential role in host plant-pathogen interactions. They degrade the plant cuticle and tissue and enable pathogen invasion. Such enzymes, specifically cutinase and pectinase, have been considered potential bio-scouring agents to degrade the cotton fabric cuticle’s outer layer at low temperature and alleviate environmental pollution. In this work, the combined effect of cutinase, pectin lyase, or polygalacturonase on the scouring of cotton fabrics was studied using evaporative light-scattering reverse-phase HPLC and GC-MS analysis of the reaction components, and measuring changes in the cotton fabrics’ properties. The traditional method of cotton fabrics’ scouring with NaOH resulted in decreased pectin content and increased cellulose fibers accessibility, evaluated by specific staining. Treating the cotton fibers’ cuticle with cutinase led to the acidification of the reaction mixture, a decrease in enzyme-specific activity, and elevation in hexadecanoic acid and octadecanoic acids in the reaction fluid. These two saturated fatty acids are the main wax constituents of raw cotton fabrics, identified using GC-MS after dichloromethane reflux overnight. Treating cotton fabrics with each of the three enzymes, cutinase, pectin lyase, or polygalacturonase, increased their pectin removal, as measured by high concentrations of D-galacturonic acid and other pectin constituents in the reaction fluid. A synergistic effect was found in the combined treatment of cutinase and pectin lyase in the hydrolysis of the cotton fibers’ cuticle. This effect was expressed in high water absorbency of the treated fibers, increased fabric weight loss and sharp elevation of a cutin and pectin monomer’s related peaks (retention time [RT] = 4.1 min and 2.9, 4.5 min, respectively). A model was suggested for the synergistic action between cutinase and pectin lyase. It assumes that the cuticle’s digestion by cutinase results in the enlargement and formation of outer layer micropores, which enables the rapid penetration of pectinase into the inner pectin layer.