Hydrolytic bacteria have the ability to convert organic material which is very useful in the bioremediation of biomedical waste. The objective of the study was to investigate the hydrolytic bacterial diversity that has the hydrolase enzyme activity. The medical waste sampling was carried out at Roemani Hospital and KRMT Wongsonegoro Hospital, Semarang Central Java in mid-August 2016. Microbiological and molecular studies, covering isolation and purification, hydrolase productivity tests, and bacterial determination, were conducted at the Tropical Marine Biotechnology Laboratory, Faculty of Fisheries and Science, UNDIP. The 16S rDNA gene sequence analysis was carried out by using the BLAST GeneBank NCBI homology test, while the phylogenetic tree construction was carried out by means of Clustal X software and Phylegenetic analysis using parsimony (PAUP vers 4.0). The test results showed that 26 hydrolytic bacterial isolates were able to produce four types of extracellular hydrolase enzymes. The results of the enzyme production test showed that there were only 2 bacterial isolates capable of producing the four types of enzymes. Microbiological and molecular polyphasic identifications showed that the 2 isolates, R1-17, and R2-6 isolates are closely related to Virgibacillus salarius strain B-11 and Bacillus subtilis strain VITVB1, respectively. Even though there are only 2 isolates capable of producing the 4 types of extracellular enzymes, it seems that hospital biomedical waste is a potential source for obtaining hydrolytic bacteria.