In this study, the antioxidant activity, antimicrobial effect, and fatty acid composition of the bark of the Turkish Pine (Pinus brutia Ten.) grown in Burdur Province (Turkiye) were investigated. Red pine bark extracts were analyzed using the Folin-Ciocalteau method for total phenolic content, the DPPH test for free radical scavenging activity, the FRAP test for reduction power, and the carotene-linoleic acid test to determine antioxidant activity. Red pine bark was found to have a significant antioxidant effect. The total phenomenon, expressed as gallic acid, was found to be 287.31 mg GAE/100 g, while the free radical inhibition rate was found to be 93.94%. The results were compared with the standard antioxidants BHA and BHT. The antimicrobial activity of the chloroform, methanol, and ethyl acetate extracts of red pine bark was examined against four bacteria strains using the microdilution method, and their fatty acid compositions were determined using a GC-FID device. Although it showed no activity against Staphylococcus aureus, it produced remarkable results against Bacillus cereus and Escherichia coli in particular. Therefore, it was found to have a good antibacteria effect and an important fatty acid profile for industry with long-chain (24-carbon) saturated and long-chain unsaturated fatty acids. While the SFA value was high (89.23%), C24:0 (lignoceric acid) was the predominant fatty acid with a proportion of 51.5%, followed by C22:0 at 28.38%.