Water activity (aw) as an important parameter for self-preservation can help to control microbial growth in cosmetic formulations. However, high amounts of water-binding substances are required to lower the aw enough to affect microbial growth. Since consequences for the skin barrier have been poorly studied so far, we investigated the effect of aw-lowering agents on both the antimicrobial properties of o/w emulsions and skin physiological parameters. A combination of selected natural humectants (Sodium lactate, Propanediol, Erythritol, Betaine and Sodium PCA) with a total concentration of 28 wt% in an o/w emulsion was able to reduce its aw from 0.980 ± 0.003 to 0.865 ± 0.005. The challenge test results of the aw-lowered emulsion showed a convincing microbial count reduction in potentially pathogenic microorganisms. The addition of as little as 0.5% of the antimicrobial multifunctionals Glyceryl Caprylate and Magnolia Officinalis Bark Extract further enhanced the antimicrobial effect, resulting in adequate antimicrobial protection. Moreover, twice-daily application of the aw-lowered emulsion for a period of four weeks led to a skin barrier-enhancing effect: TEWL significantly decreased, and SC hydration significantly increased. Thus, we present an opportunity to replace conventional preservatives with a natural alternative preservation strategy that has been shown to offer benefits for the skin.