Molecular characterization and secreted production of basidiomycetous cell-bound β-glycosidases applicable to production of galactooligosaccharides.

Cell-bound β-glycosidases of basidiomycetous yeasts show promise as biocatalysts in galactooligosaccharide (GOS) production. Using degenerated primers designed from Hamamotoa singularis (Hs) bglA gene, we newly identified three genes that encode cell-bound β-glycosidase from Sirobasidium magnum (Sm), Rhodotorula minuta (Rm), and Sterigmatomyces elviae (Se). These three genes, also named bglA, encoded family 1 glycosyl hydrolases with molecular masses of 67‒77 kDa.

Probiotics into outer space: feasibility assessments of encapsulated freeze-dried probiotics during 1 month's storage on the International Space Station.

Suppression of immune function during long spaceflights is an issue that needs to be overcome. The well-established probiotic Lactobacillus casei strain Shirota (LcS) could be a promising countermeasure, and we have launched a project to investigate the efficacy of its use on the International Space Station (ISS). As a first step, we developed a specialist probiotic product for space experiments, containing freeze-dried LcS in capsule form (Probiotics Package), and tested its stability through 1 month of storage on the ISS.

Functional analysis of a novel hydrogen peroxide resistance gene in Lactobacillus casei strain Shirota.

Lactic acid bacteria have a variety of mechanisms for tolerance to oxygen and reactive oxygen species, and these mechanisms differ among species. Lactobacillus casei strain Shirota grows well under aerobic conditions, indicating that the various systems involved in oxidative stress resistance function in this strain. To elucidate the mechanism of oxidative stress resistance in L.

Expression of Porphyromonas gingivalis gingipain antigen Hgp44 domain on surface of Lactococcus lactis.

Porphyromonas gingivalis, a pathogen involved in the development of chronic periodontitis, has a number of major virulence factors, among which are its surface cysteine protease gingipains. The purpose of this study was to investigate the feasibility of inducing protective antibodies against P. gingivalis by means of immunization with recombinant Lactococcus lactis expressing the 44-kDa gingipain adhesion/hemagglutinin domain (Hgp44).

Transposon mutagenesis of probiotic Lactobacillus casei identifies asnH, an asparagine synthetase gene involved in its immune-activating capacity.

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.

A practical random mutagenesis system for probiotic Lactobacillus casei using Tn5 transposition complexes.

Aims: Despite the fact that the entire genome sequence of probiotic Lactobacillus casei has recently been available, their mechanisms of beneficial effects are poorly clarified, probably because of the lack of an efficient mutagenesis system. The aim of this study was to establish a practical random mutagenesis system of L. casei using the Tn5 transposome complexes.

Antimicrobial susceptibility of Bifidobacterium breve strains and genetic analysis of streptomycin resistance of probiotic B. breve strain Yakult.

The minimum inhibitory concentrations (MICs) of 17 antimicrobials for 26 Bifidobacterium breve strains of various origins were determined by broth microdilution. MIC distributions for 17 antimicrobials except streptomycin and tetracycline were unimodal for all strains tested, whereas bimodal distributions were observed for streptomycin and tetracycline. The probiotic strain B.

Immunogenicity and protective efficacy of orally administered recombinant Lactococcus lactis expressing surface-bound HIV Env.

This study investigates whether genetically modified orally administered Lactococcus lactis (L lactis) could be used as an HIV vaccine. L lactis is immunogenic and extremely safe when delivered orally. We created a recombinant L lactis vector expressing the envelope protein of HIV on its cell surface.

Insertion of bacteriophage phiFSW into the chromosome of Lactobacillus casei strain Shirota (S-1): characterization of the attachment sites and the integrase gene.

The integrase gene (int) on the genome of φFSW, which is a temperate bacteriophage of Lactobacillus casei strain Shirota (formerly denoted as S-1), and the four attachment sites on the genomes of the phage and its host were characterized by sequencing. The φFSW integrase was found to belong to the integrase family of site-specific tyrosine recombinase. The attachment sites shared a 40bp common core within which an integrative site-specific recombination occurs.

Molecular characterization of a cell wall-associated proteinase gene from Streptococcus lactis NCDO763.

Streptococcus lactis NCDO763 harbours a plasmid designated pLP763. The cells harbouring pLP763 are able to grow to a higher density in milk because of their proteinase-positive phenotype (Prt+). The 6.

ISL1: a new transposable element in Lactobacillus casei.

The genome structures of a temperate Lactobacillus phage, phi FSW, and its virulent mutants, phi FSVs, were examined by restriction, heteroduplex and nucleotide-sequence analyses. The results showed that two out of three phi FSVs had the same 1.3 kbp insertion (designated as ISL1) at different positions in the phi FSW sequence.

BinI: a new site-specific endonuclease from Bifidobacterium infantis.

A new restriction endonuclease, BinI, from Bifidobacterium infantis 659 has been isolated. By mapping and sequencing of the cleavage sites on SV40 DNA, it was deduced that BinI recognizes the asymmetric pentanucleotide sequence (formula; see text) and cleaves at the sites indicated by the arrows, generating mononucleotide 5'-terminal extensions. BinI is a member of a unique class of Type-II restriction endonucleases which recognize a specific but asymmetric sequence and cleave at a site several bases away.

Two site-specific endonucleases BinSI and BinSII from Bifidobacterium infantis.

Two site-specific endonucleases, BinSI and BinSII, were isolated from Bifidobacterium infantis S76e. BinSI was found to be an isoschizomer of EcoRII, while BinSII was shown to have the same sequence and cutting specificity as BbeI, 5'-GGCGC decreases C-3'. Both BinSII- and BbeI-generated DNA fragments could be ligated with HaeII-generated DNA fragments.