Concomitant production of lipase, protease and enterocin by Enterococcus faecium NCIM5363 and Enterococcus durans NCIM5427 isolated from fish processing waste

Enterococci are widely distributed in the environment ranging from foods to humans and are gaining industrial importance due to their technological traits. In the present study, enterococci (Enterococcus faecium NCIM5363 (EF-63) and Enterococcus durans NCIM5427 (ED-27)) which are native to fish processing waste with an ability to produce lipase, protease and enterocin concomitantly were characterised. Lipase assay was performed by titrimetry and protease activity and was estimated using haemoglobin and casein as substrates in the presence of buffers at acidic, basic and neutral pH. Furthermore, enterocin produced by the isolates was characterised. Enterocin was also checked for its stability at different pH, temperature and proteolytic enzymes. Lipase production was found to be 22 and 10 U/ml in the absence of tributryin and increased to 40 and 24 U/ml in its presence for EF-63 and ED-27, respectively, indicating that the lipase produced is substrate dependent. Protease production was confirmed by protease assay, and the protease produced showed more affinity towards the acidic substrate. Enterocin produced was stable at low pH (2 to 3) and high temperature (121°C, 15 min) and had a molecular weight of approximately 6 kDa. It exhibited antibacterial activity against both Gram-positive and Gram-negative food-borne pathogens. Proteinase K inactivated enterocin completely, whereas trypsin did not. Novelty of this work lies in the immense industrial importance these cultures hold as they are capable of producing lipase, protease and enterocin apart from being useful in recovering proteins and lipids from fish processing wastes.