A new drug delivery system (DDS) was prepared by electrospinning and multi-layered coating methods. Lactobacillus was used as a target drug for release in the large intestine. Lactobacillus was incorporated into polyvinyl alcohol-based electrospun fibers. Alginate and chitosan were then used to coat these fibers to protect and deliver lactobacillus to the large intestine. The multi-layered coating procedure was carried out at various coating times to determine the optimal coating amount in human digestion conditions. The drug-release experiment was conducted using an in vitro model of human digestion processes. Chitosan and alginate were released by dissolving in acid and neutral conditions, respectively. Lactobacillus was then released in a large intestine model by the hydrogel swelling effects of PVA-based electrospun fibers. The drug-release behavior was predicted by calculated equations for optimized coating times. Eventually, the lactobacillus was effectively delivered to the target large intestine by chitosan and alginate coating and was released by PVA hydrogel swelling. (C) 2011 The Korean Society of Industrial and Engineering Chemistry. Published by Elsevier B.V. All rights reserved.