Fujitoshi Yanagida

Isolation and identification of lactic acid bacteria from soil using an enrichment procedure

Aims:  To survey, and identify and classify the ecological distribution of lactic acid bacteria from soil in Japan and Taiwan.

Distribution of phenolic yeasts and production of phenolic off-flavors in wine fermentation

The activity of wine yeasts to decarboxylate ferulic and p-coumaric acids is one of their biological properties related to the production of phenolic off-flavors (POF) in wine-making. We examined POF productivity in 116 strains of wine yeast, 74 strains of wild yeast (Saccharomyces cerevisiae) and 23 strains of non-Saccharomyces yeast, and found that a majority of these yeasts were POF-producing strains. The frequency distribution of POF-producing strains was 81 to 95% in wine yeasts, 85 to 97% in wild yeasts and 78 to 83% in non-Saccharomyces yeasts based on the POF test with addition of ferulic and p-coumaric acids to grape juice medium. The Rhodotorula, Candida, Cryptococcus, Pichia, Hansenula, and Brettanomyces strains had high or moderate POF productivity among the 20 non-Saccharomyces species. The decomposition rate of ferulic acid correlated with POF production and the critical concentration of phenolic acid (free form) in grape must was estimated to be more than 10 mg/l. Segregation of POF phenotype and Southern blot analysis of phenolic wine yeasts suggest that POF production is controlled by the POF gene (PAD1). The results showed the frequent distribution of phenolic yeasts in the wine-making environment. These suggest the importance of controlling POF production by using wine yeast strains of low POF productivity. The grapes must be prepared by a suitable process to prevent the increase in phenolic acid content.

Weissellicin 110, a Newly Discovered Bacteriocin from Weissella cibaria 110, Isolated from Plaa-Som, a Fermented Fish Product from Thailand

ABSTRACT Weissella cibaria 110, isolated from the Thai fermented fish product plaa-som, was found to produce a bacteriocin active against some gram-positive bacteria. Bacteriocin activity was not eliminated by exposure to high temperatures or catalase but was destroyed by exposure to the proteolytic enzymes proteinase K and trypsin. The bacteriocin from W. cibaria 110 was purified, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the purified bacteriocin contained one protein band that was approximately 2.5 kDa in size. Mass spectrometry analysis showed the mass of the peptide to be approximately 3,487.8 Da. N-terminal amino acid sequence analysis was performed, and 27 amino acids were identified. Because it has no similarity to other known bacteriocins, this bacteriocin was defined as a new bacteriocin and termed weissellicin 110.

Durancin L28‐1A, a new bacteriocin from Enterococcus durans L28‐1, isolated from soil

Aims:  To isolate, characterize and identify bacteriocins from lactic acid bacteria in soil.

Production and purification of a bacteriocin peptide produced by Lactococcus sp. strain GM005, isolated from Miso-paste.

Lactococcus sp. GM005 was isolated from Miso-paste and was found to produce a bacteriocin with strong antibacterial activity. A culture of Lactococcus sp. GM005, maintained at 30 degrees C and a constant pH of 6.0, exhibited bacteriocin activity eightfold higher than that of a culture grown under pH-uncontrolled conditions. GM005 bacteriocin was purified to homogeneity on SDS-PAGE by hydrophobic column chromatography and gel filtration. The estimated molecular weight of GM005 bacteriocin was approximately 9.6 kDa based on gel-filtration analysis, and was approximately 2.4 kDa based on tricine-SDS-PAGE analysis, indicating a tetrametric structure. N-terminal amino acid analysis revealed that the N-terminal end was blocked. Amino acid composition analysis revealed a high proportion of hydrophobic amino acid residues and lanthionine. This differs from the composition of some antibiotics. GM005 bacteriocin exhibits a bactericidal activity against Lactobacillus sakei JCM1157T.

Restriction fragment length polymorphism analysis of 16S rRNA genes in lactic acid bacteria isolated from red wine

Various lactic acid bacteria isolated from wine and alcoholic drinks attempted to identified by restriction fragment length polymorphism (RFLP). Oenococcus oeni strains exhibited unique RFLP patterns by HaeIII-digestion of 12 reference strains. We performed RFLP analysis using the AccII or HaeIII enzyme for 44 strains isolated from red wine and the results indicated profiles identical to O. oeni type strain. O. oeni does not exhibit interspecific diversity. Thus, the use of RFLP analysis of 16S rDNA is useful in the identification of O. oeni strains isolated from wines.

Isolation and characteristics of lactic acid bacteria from koshu vineyards in Japan

Aims:  To isolate, characterize and identify lactic acid bacteria (LAB) in the vineyards where koshu grapes, a primary wine grape cultivar in Japan, are grown.

Effects of prebiotic oligosaccharides and trehalose on growth and production of bacteriocins by lactic acid bacteria.

Aims:  To investigate the effects of two prebiotics and trehalose on the production of bacteriocins.

Proposal of Sporolactobacillus nakayamae subsp. nakayamae sp. nov., subsp. nov., Sporolactobacillus nakayamae subsp. racemicus subsp. nov., Sporolactobacillus terrae sp. nov., Sporolactobacillus kofuensis sp. nov., and Sporolactobacillus lactosus sp. nov.

Catalase-negative spore-forming lactic acid bacteria were isolated from soil samples and fermentation starters for Asian traditional alcoholic beverages. The isolates were characterized by determining morphological, biochemical, physiological, and chemotaxonomic properties and were found to be members of the genus Sporolactobacillus. Twelve isolates and some authentic strains belonging to this genus were used in DNA base composition and DNA relatedness studies, and the results revealed that the strains tested could be divided into six groups which correlated with the phenotypic characteristics. One of the groups corresponded to the previously described species Sporolactobacillus inulinus. In addition, we propose the following four new species and two new subspecies for the remaining five groups: Sporolactobacillus nakayamae subsp. nakayamae sp. nov., subsp. nov. (type strain, M-114 [= JCM 3514]), Sporolactobacillus nakayamae subsp. racemicus subsp. nov. (type strain, M-17 [= JCM 3417]), Sporolactobacillus terrae sp. nov. (type strain, M-116 [= JCM 3516]), Sporolactobacillus kofuensis sp. nov. (type strain, M-19 [= JCM 3419]), and Sporolactobacillus lactosus sp. nov. (type strain, Xl-1 [= JCM 9690]).

Isolation and characterization of lactic acid bacteria from dochi (fermented black beans), a traditional fermented food in Taiwan.

Aims:  To isolate, characterize and identify lactic acid bacteria (LAB) in dochi (fermented black beans), a traditional fermented food in Taiwan.