Novel listerial genetic loci conferring enhanced barotolerance in Escherichia coli.

ABSTRACT

AIMS: To identify Listeria monocytogenes genes with a role in high-pressure (HP) resistance. METHODS AND RESULTS: A L. monocytogenes genomic library constructed in an Escherichia coli background was screened for loci conferring increased HP resistance. Pressure treatments at 400 megapascals for 5 min in Luria-Bertani (LB) agar were used to screen for increased resistance to pressure. Colonies arising on the treated agar plates were isolated, the plasmid extracted and the inserts sequenced to identify the genetic loci conferring HP resistance. Seven different genetic regions were identified, which encoded proteins similar to an inorganic polyphosphate/ATP-NAD kinase, the septation ring formation regulator EzrA, flagellar motor proteins MotA and MotB, proteins similar to the quorum sensing Agr system from Staphylococcus (AgrA, AgrC and AgrD), proteins similar to a transcription regulator (RpiR family) and a fructose phosphotransferase system, proteins of unknown function, and a Fur regulator. Of the seven loci confirmed, three (EzrA, MotA/B and the Agr system) maintained significantly reproducible HP tolerance when expressed in a different E. coli background. CONCLUSIONS: Novel genetic loci from the L. monocytogenes genome confer increased HP resistance when heterologously expressed in an E. coli background. SIGNIFICANCE AND IMPACT OF THE STUDY Molecular and functional approaches to the screening of genetic elements linked to HP resistance provide greater insights into microbial inactivation and/or survival mechanisms when using HP as a means of controlling/eliminating bacterial growth. This information will ultimately have significant implications for the use of HP processing in the food industry, in terms of both food quality and safety.