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Autochthonous single (Ent+) or multiple (m-Ent+) enterocin-producing strains of dairy enterococci show promise for use as bioprotective adjunct cultures in traditional cheese technologies, provided they possess no pathogenic traits. This study evaluated safety, decarboxylase activity, and enzymatic (API ZYM) activity profiles of nine Ent+ or m-Ent+ Greek cheese isolates previously assigned to four distinct  E. faecium ( represented by the isolates KE64  (entA),  GL31  (entA) , KE82  (ent A -ent B -ent P) and KE77  (ent A -ent B -ent P -bac31) ) and two  E. durans  (represented by the isolates KE100  (ent P )  and KE108  (ent P -bac31-cyl )) strain genotypes. No strain was  β -hemolytic or harbored  van A and  van B or the virulence genes  agg, ace, espA, IS16, hyl , or  gel E. All strains were of moderate to high sensitivity to ampicillin, ciprofloxacin, chloramphenicol, erythromycin, gentamicin, penicillin, tetracycline, and vancomycin, except for the  E. faecium  KE64 and KE82 strains, which were resistant to erythromycin and penicillin. All cheese strains showed moderate to strong esterase-lipase and aminopeptidase activities and formed tyramine, but none formed histamine in vitro. In conclusion, all Ent+ or m-Ent+ strain genotypes of the  E. faecium/durans  group, except for the  cyl -positive  E. durans  KE108, were safe for use as adjunct cultures in traditional Greek cheeses. Further in situ biotechnological evaluations of the strains in real cheese-making trials are required.

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Safety Evaluation, Biogenic Amine Formation, and Enzymatic Activity Profiles of Autochthonous Enterocin-Producing Greek Cheese Isolates of the Enterococcus faecium/durans Group