Dehydrins are a group of plant proteins that have been shown to be involved in the tolerance of various abiotic stresses such as dehydration, salinity, and low temperature. We have previously shown that the K-segments of the wheat dehydrin DHN-5 are essential for the protection of enzyme activities in vitro. In this study, we further investigate the role of the K-segments in the growth of Escherichia coli under various stresses, and we tested their antibacterial and antifungal activities. Our results showed that the truncated forms of DHN-5 containing the two K-segments enhanced tolerance of E. coli against diverse stresses by protecting proteins against aggregation. In addition, we demonstrated that the K-segments have antibacterial and antifungal activities against Gram-positive and Gram-negative bacteria and fungi. Based on these results, we propose that the K-segments may play a protective role in plants not only under abiotic stress conditions but also most likely during defense mechanisms.