Thyme, mint, and lemon balm were used to determine whether shading conditions could improve the yield, composition, antioxidant, and antimicrobial activity in plant essential oils (EOs) in comparison with non-shaded plants from an open field. The yield of the EOs of non-shaded thyme, mint, and lemon balm, was 3.44, 3.96, and 0.21 mL/100 g, respectively. Plants covered by nets produced different levels of EOs (3.46, 2.20, and 0.45 mL/100 g) after 120 min of hydrodistillation. The main components of the thyme essential oil are thymol (44.2–43.9%), γ-terpinene (18.3–16.8%), and p-cymene (16.5–17.4%). The predominant components of mint essential oil are piperitenone oxide (52.6–64.8%) and 1,8 cineole (25.9–16.3%), while lemon balm essential oil consists of the following main components: geranial (34.0–32.8%); neral (21.3–24.9%); and piperitenone oxide (17.2–16.7%). The EOs from non-shaded thyme and mint plants have the highest antioxidant activity (EC50 value 0.54 mg/mL and 3.03 mg/mL). However, shaded lemon balm showed a stronger antioxidant activity (EC50 3.43 mg/mL) than non-shaded plants (12.85 mg/mL) after 60 min of incubation. The EOs from all plants showed significant effects against Escherichia coli, Proteus vulgaris, Bacillus subtilis, Staphylococcus aureus, and Candida albicans. The most active EOs against most of the isolates originated from Thymus vulgaris L., plants. Adequate cultivation techniques, such as shading for Lamiaceae plants, has positive effects, especially in Melissa officinalis L. Shading can achieve a higher content and components in terms of the specific biological activity (antioxidant and microbial) of EOs.