Actinomycete strain VY31 was isolated from agriculture soil of region Východná, Slovakia. Morphological, physiological and biochemical studies indicated that this isolate belongs to the genus Streptomyces. The 16S rRNA sequence data supported the assignment of the isolate to the genus Streptomyces rishiriensis (sequence similarity 97%). Tested isolate was able to produce melanin dark pigment and exopigments on ISP6, ISP7 and SSM+T cultivating media. The optimal pH range was from 6-8 and optimal temperature at 30 °C. The strain exhibited salt tolerance up to 5 % and utilized the carbon sources such as glucose, arabinose, xylose, inositol, mannose, fructose, rhamnose and rafinose. Using ApiZym® stripes, the highest production of enzymes was determined for phosphatase alkaline, leucinearylamidase, valinearylamidase, phosphatase acid, naphtol-AS-BI-phosphohydrolase, galactosidase and glucosidase (>40 nmol). According to ApiCoryne® results, positive reaction was confirmed in case of esculin, alkaline phosphatase, and this strain was also able to hydrolyze gelatine. Minimum Inhibitory Concentration (MIC) of the purified extract of isolate was evaluated against Gram-positive bacteria Staphylococcus aureus and Enterococcus faecium, Gram-negative bacteria Escherichia coli and Pseudomonas aeruginosa and against yeast Candida albicans. On the basis of MIC results, strain VY31 had noticeable antibacterial activity against Staphylococcus aures N315 (MRSA) from collection database of University Hospital in Hamburg, Germany. This isolate could be used in the development of new antibiotics for pharmaceutical purposes.