This investigation reports a new biotechnological process that uses L-cysteine (L-Cys) which provides a permanent, nontoxic and effective antimicrobial effect over wool-based materials. This process is simple and carried out via widespread exhausting methods. Typically, wool fabrics are incubated with L-Cys for 50 min at 60 degrees C in a pH 4.8 acetate buffer solution 25 mM, under mild agitation to give a good absorption rate. The minimal inhibitory concentration (MIC) of L-Cys was evaluated by the NCCLS M07-A6 standard method, and the results showed a good antibacterial activity against S. aureus and K. pneumoniae within the range of 6.0 x 10(-3) - 4.8 x 10(-2) g/mL [MIC 0.6% (w/v)] and 6.0 x 10(-3) - 4.8 x 10(-2) g/mL [MIC 0.6% (w/v)], respectively. In addition, the antimicrobial activity of the functionalized wool was assessed by the international standard JIS 1902-2002 showing a good inhibition of bacterial growth for an L-Cys concentration of 1% over the weight of fabric, both against Staphylococcus aureus and Klebsiella pneumoniae. Moreover, the biocidal mechanism was found to be related with the increase of sulfhydryl's groups onto wool fibers, which were quantified by the Ellman's reagent (5,5'-Dithio-bis(2-nitrobenzoic acid) method. The new process is easy to perform, non toxic, preserve wool quality and is a novel biomimetic approach that uses antimicrobial amino acids and may open new avenues for the design of biomedical textiles with a broad range of applications in healthcare. (C) 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2012