The purpose of this study is to examine the antimicrobial and antifungal activity of propolis samples of Yigilca honey bee against Gram-positive and Gram-negative bacteria and yeast-like fungi. For this purpose the pollen and chemical components analyses of propolis sample was performed. The Liquid Chromatography-Mass Spectrometry (LC-MS/MS) method was used to identify the compounds of the propolis samples. According to the results of pollen analysis Fagaceae family were the dominant. As a result of LC-MS/MS method the main components of propolis was pinostrobin, gallic acid, and with Syringic acid. The antimicrobial activity of the propolis samples were tested against 7 Gram (+), 4 Gram (-) and 1 acid-resistant bacteria, and 3 yeast-like fungi. First, the activity of the propolis sample against these microorganisms was determined by the agar well diffusion method, then their zones were measured. The macrodilution method was used to determine the minimum inhibitory concentration (MIC) for the antimicrobial activity tests. The results of MIC values were found between 74 and 4746 mu L for the tested microorganisms. It was found that Mycobacterium smegmatis was the most susceptible bacteria (74 mu L), Lactobacillus acidophilus (149 mu L) Staphylococcus aureus and Saccharomyces cerevisiae with a MIC concentration of 299 mu L. We are opinion that, considering the biological effects of the propolis extract of Yigilca honey bee, may be potentially used as an alternative therapeutic treatment thanks to its high antimicrobial and antifungal activity on different microorganisms, but its high in vitro activity needs to be confirmed by further research in vivo.