An electroporation‐induced transformation system was developed for two raw sausage starter organisms: Lactobacillus curvatus Lc2‐c and Lact. sake Ls2. Several plasmids of different origin (pAMβ1, pIL253, pNZ12 and pLP825) were tested for transformation and expression of their resistance determinants. Transfer rates were strongly dependent on growth phase and voltage. Several parameters were studied with possible influence on trasformation efficiencies.

Isolates of lactobacilli from infant faeces phenotypically characterized as Lactobacillus paracasei subsp. paracasei (six strains), Lact. rhamnosus (six strains), Lact. gasseri (three strains), Lact. acidophilus (one strain) and Lact. fermentum/reuteri (three strains) according to recent classification systems were subjected to SDS-PAGE of whole cell proteins and rRNAtargeted oligonucleotide probe hybridization, in order to confirm the phenotypic characterization and elucidate the exact taxonomic position of the three strains that had properties between fermentum and reuteri. Results suggested a good agreement between the phenotypic characterization, SDS-PAGE and rRNA-targeted oligonucleotide probe hybridization for strains of all species except for the Lact. fermentum/reuteri strains. Results obtained by rRNA probes suggested a possible phylogenetic relatedness of the strains to Lact. reuteri. Isolates from infant faeces with interesting probiotic properties could be used as components of fermented milk products.

The lysostaphin structural gene was cloned in Bacillus subtilis DSM402 and in Lactobacillus casei 102S. The gene was expressed in both organisms and active lysostaphin was released into the medium. Lysostaphin produced by these organisms induced lysis of growing and heat inactivated staphylococci. Expression in a protective starter organism is a prerequisite to produce lysostaphin in situ in fermenting foods and hence, to reduce the hygienical risk of staphylococcal food poisoning.

No abstract

An electroporation based transformation system was developed for Lactobacillus curvatus Lc2-c, a plasmid cured potential raw sausage starter. It was transformed with plasmid pLipPS1, a staphylococcal vector plasmid containing the lipase gene from Staphylococcus hyicus. Transformants exhibited high lipase activity, whereas the wild type strain was lipase negative. The production and release into the medium of a heterologous lipase by a Lactobacillus could be shown for the first time.