Protocol for Initial Purification of Bacteriocin.

摘要

This report describes a protocol designed to instruct on how to partially purify an unknown bacteriocin to allow for rapid evaluation of its potential as an effective antimicrobial, and it briefly summarizes the effort by the Natick Soldier Research, Development and Engineering Center (NSRDEC), between February 2013 and August 2014, to develop the protocol. Bacteriocins are narrow-spectrum toxins that control pathogenic bacteria by out-competing them for resources while leaving beneficial bacteria unaffected. One of the limiting factors for the commercialization of bacteriocins is the time consuming and costly process for evaluating an unknown a bacteriocin for its potential effectiveness, which involves obtaining an ultra pure material and then characterizing and evaluating the bacteriocin. The aim of the NSRDEC effort was to develop a purification protocol that could be applied to each isolated bacteriocin to purify the material in sufficient quantities and to a sufficient purity to make initial evaluations. The new NSRDEC protocol includes filtering the lysate/filtrate through filters with a 30,000 molecular weight cut-off (MWCO), 10,000 MWCO, and 1,000 MWCO pore size to remove unwanted protein/debris. These filtering steps are performed before the extract is loaded onto the column. Evaluation of the new protocol indicated the filters significantly increased the purity to a level sufficient to allow essential assessment of the bacteriocin s potential for use as an effective antimicrobial.