Lipolysis within single culture and co-culture biofilms of dairy origin

Bacteria in raw milk can produce heat-stable lipases, which survive pasteurisation and subsequently reduce the shelf life of dairy products because of their ability to break down the milk fat and increase rancidity. In this study, four bacteria, originating from the surfaces of raw milk transport tankers, and a known lipase-producing bacterium were evaluated for their ability to produce lipolysis in planktonic and biofilm cultures. Lipolysis was determined using two separate assays that measured hydrolysis of the ester p-nitrophenol palmitate (pnpp) and the lipid tributyrin. The hydrolysis of pnpp per CFU within biofilms and planktonic cultures ranged from 0.01 to 8.35 and 0.01 to 0.07 nU/CFU respectively. The amount of butyric acid released from hydrolysis of tributyrin per CFU within biofilms and planktonic cultures ranged from 0.1 to 1110.3 and 0.1 to 0.3 ng/CFU, respectively. The hydrolysis of pnpp and tributyrin per CFU within biofilms was at least 10 times higher compared with the corresponding planktonic cultures. This is the first study to show that lipolysis occurs within biofilms of bacteria that were originally isolated from the surfaces of raw milk tankers. This is relevant to the dairy industry, highlighting the importance of eliminating biofilms on milk tanker surfaces as a source of heat-stable lipases.