Production and Purification of MccP136, a cold-adapted antimicrobial produced by Serratia proteamaculans 136

SANCHEZ L. A,; SIÑERIZ F.; DELGADO O.D.

Toronto, Canada

Congreso; SIM Annual Meeting and Exhibition - Industrial Microbiology and Biotechnology; 2009

Society for Industrial Microbiology SIM

Antimicrobial compounds are widespread in life forms to mediate competition, and their industrial production could be important for the potential use as preservatives in food, cosmetics and in pharmaceutical industries. Cold environments could be a suitable source of microorganisms with ability to produce cold-active antimicrobials of biotechnological interest.A psychotropic Serratia proteamaculans 136 strain producing MccP136 was isolated from soil samples from Isla de los Estados (Tierra del Fuego, Argentina). MccP 136 production was optimized and a maximum of 800 AU ml-1 were obtained at early-stationary phase of growth in batch fermentations carried out in a BIOFLO 101 bioreactor with a working volume of 8.1 l with pH, temperature and dissolved oxygen control. The compound was active in a broad range of pH values (1 to 13) and presented an inhibitory effect against both Gram (+) and Gram (-) microorganisms. A purification procedure was designed including activated charcoal adsorption, solid phase extraction (SPE) and RP-HPLC under semi-preparative conditions. The purity of the compound was checked by analytical RP-HPLC and monitored by photodiode array detection and purification parameters were calculated in each purification step. The molecular mass of the antimicrobial compound (480.2 g/mol) was determined in a Hybrid Quadrupole-TOF LC/MS/MS Mass Spectrometer by ion analysis. In conclusion, MccP136, a new cold-active antimicrobial is active against diverse pathogenic strains, known as etiological agents of common infections and food-borne diseases and it could be important for applications in products that involve “cold-steps” during manufacture or processing.