Hydrolyzed milk medium for growth of some lactic acid bacteria (Streptococcus salivarius subsp. thermophilus and Lactococcus lactis), Bacillus subtilis and yeast Saccharomyces cerevisiae was developed. In this study we have attempted to improve an alternative growth medium for these bacteria and yeast instead of their conventional media. The growth rates of these microorganisms in hydrolyzed milk medium had small amount lower than their conventional media in general. Therefore the investigation suggests that hydrolyzed milk medium can be used for microbiological and genetic studies.