Insertion of bacteriophage φFSW into the chromosome of Lactobacillus casei strain Shirota (S-1): Characterization of the attachment sites and the integrase gene
“…However, YIT 9029 bound only to CSL, a rhamnose-specific lectin (32,33,37). It is known that PS from YIT 9018, the parental strain of YIT 9029 which was produced by removing bacteriophage FSW from the YIT 9018 genome (33), contains Rha, Glc, Gal, N-acetylglucosamine (GlcNAc), and Nacetylgalactosamine (GalNAc) (28).…”
The patterns of lectin-binding affinity of most strains were found to be unique. There appears to be two types of lectin-binding profiles: the first is characterized by a few lectins, and the other is characterized by multiple lectins with different specificities. We also showed a dramatic change in the lectin-binding profile of a YIT 9029 derivative with a mutation in the cps1C gene, encoding a putative glycosyltransferase. In conclusion, the developed technique provided a novel strategy for rapid profiling and, more importantly, differentiating numerous bacterial strains with relevance to the biological functions of PS.
“…However, YIT 9029 bound only to CSL, a rhamnose-specific lectin (32,33,37). It is known that PS from YIT 9018, the parental strain of YIT 9029 which was produced by removing bacteriophage FSW from the YIT 9018 genome (33), contains Rha, Glc, Gal, N-acetylglucosamine (GlcNAc), and Nacetylgalactosamine (GalNAc) (28).…”
The patterns of lectin-binding affinity of most strains were found to be unique. There appears to be two types of lectin-binding profiles: the first is characterized by a few lectins, and the other is characterized by multiple lectins with different specificities. We also showed a dramatic change in the lectin-binding profile of a YIT 9029 derivative with a mutation in the cps1C gene, encoding a putative glycosyltransferase. In conclusion, the developed technique provided a novel strategy for rapid profiling and, more importantly, differentiating numerous bacterial strains with relevance to the biological functions of PS.
“…L. casei ATCC 27139, the parental strain of Shirota [15], has been consistently shown to exhibit anti-infectious activity against L. monocytogenes and anti-tumor activity against MethA fibrosarcoma in mice associated with the marked increases in the cytokine levels such as TNF-α, IL-12, IL-18, and IFN-γ [16]. Although L. casei ATCC 27139 is susceptible to J1 phage, which shows a remarkably high specificity to certain strains of L. casei/L.…”
Lactobacillus casei ATCC 27139 enhances host innate immunity, and the J1 phage-resistant mutants of this strain lose the activity. A transposon insertion mutant library of L. casei ATCC 27139 was constructed, and nine J1 phage-resistant mutants out of them were obtained. Cloning and sequencing analyses identified three independent genes that were disrupted by insertion of the transposon element: asnH, encoding asparagine synthetase, and dnaJ and dnaK, encoding the molecular chaperones DnaJ and DnaK, respectively. Using an in vivo mouse model of Listeria infection, only asnH mutant showed deficiency in their ability to enhance host innate immunity, and complementation of the mutation by introduction of the wild-type asnH in the mutant strain recovered the immuno-augmenting activity. AsnH protein exhibited asparagine synthetase activity when the lysozyme-treated cell wall extracts of L. casei ATCC 27139 was added as substrate. The asnH mutants lost the thick and rigid peptidoglycan features that are characteristic to the wild-type cells, indicating that AsnH of L. casei is involved in peptidoglycan biosynthesis. These results indicate that asnH is required for the construction of the peptidoglycan composition involved in the immune-activating capacity of L. casei ATCC 27139.
Lysogeny entails more economical and technological risks in probiotic Lactobacillus casei/paracasei bacteria than in other lactic acid bacteria, due to economic value. Lysogeny is widely spread among L. casei/paracasei strains. Siphophages CL1 and CL2, isolated from noninfected lysed‐cultures of commercial L. paracasei A, are thermoresistant, have identical host spectrum, latent and burst times, whereas burst sizes are 148 and 85, respectively. Mitomycin C induction of L. paracasei A yielded prophage iA2, whose presence was confirmed on the strain genome and whose restriction patterns differed from phages CL1 and CL2. The latter shared several restriction fragments, probably indicating a common origin.
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