Pseudomonas spp. plays an important role in milk spoilage. During the storage of raw milk they produce many thermo-tolerant lipolytic and proteolytic enzymes that reduce both the quality and shelf life of processed milk. This study was performed to isolate, characterize biochemical properties, determine proteolytic and lipolytic activities of Pseudomonas spp. and identify the strongest proteolytic and lipolytic activities Pseudomonas spp. by 16S rRNA genes sequencing. 14 bacterial isolates belonging to Pseudomonas spp. were isolated from 10 raw milk samples from the cow-milk stations and cow-milk farms from Tien Giang, Can Tho, Soc Trang provinces in Mekong Delta, Vietnam. Among 14 isolates, 9 isolates had proteolytic activity, while 10 isolates had lipolytic activity and 8 isolates had both. CT3 isolate had the strongest proteolytic activity and was 100 % of identify with Pseudomonas putida strain BASUP8716S rDNA. TG3 isolate had the strongest lipolytic activity and was 100 % of identify with Pseudomonas putida strain Seab04 16S rDNA.