Food preservative potential of gassericin A‐containing concentrate prepared from cheese whey culture supernatant of Lactobacillus gasseri LA39

Gassericin A (GA) is a circular bacteriocin produced by Lactobacillus gasseri LA39. In this study, GA‐containing concentrate was prepared using a cross‐flow membrane filtration device (30 kDa cut‐off) from the culture supernatant of Lb. gasseri LA39 cultivated in a cheese whey‐based food‐grade medium. The bacteriocin activity titer in the concentrate was 16 times as high as that of the culture supernatant and was completely maintained through each incubation at 4°C for 3 months, 37°C for 2 months, 60°C for 5 h, and 100°C for 30 min. The GA‐containing concentrate was used with glycine powder to make custard creams, and then four representative strains of custard cream spoilage bacteria (Bacillus cereus, Lactococcus lactis subsp. lactis, Achromobacter denitrificans and Pseudomonas fluorescens) were individually inoculated at c. 10³ colony forming units/g in the custard creams. Throughout 30 days of incubation at 30°C, all of the inoculated bacteria were completely inhibited by the combination of 5% (w/w) of the GA‐containing concentrate and 0.5% (w/w) glycine. This is the first highly practical application of GA to foods as a biopreservative, and the concentration method and the bacteriocin concentrate would contribute to biopreservation of several foods.     

Heterologous expression of gassericin A,a bacteriocin produced by Lactobacillus gasseri LA39

Gassericin A, a bacteriocin produced by Lactobacillus gasseri LA39, has a cyclic structure linking N‐ and C‐terminal amino acids. Gassericin A was expressed in Escherichia coli JM109 as a biotinylated fusion protein on the basis of the DNA sequence of mature bacteriocin. A positive clone accumulated the bacteriocin, with no activity, as a soluble fusion protein in the cytoplasm. After release of an N‐terminal tag with factor Xa protease, gassericin A was converted into an active peptide having N‐ and C‐termini. The total amount of purified bacteriocins (expressed and native) was 480 µg/L and 370 µg/L, respectively. However, the specific activity of expressed gassericin A was 15 AU/mg lower than that of native bacteriocin (2600 AU/mg). Although the actual Mr (molecular weight) of the expressed bacteriocin should be 5666, the peptide showed the same mobility (Mr 3800) in sodium dodecylsulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) as native cyclic gassericin A, suggesting that the expressed peptide retains compact folding of the molecule similar to that of native gassericin A.     

Effects of high protein levels in concentrate feed during the early fattening stage on physico-chemical composition and sensory characteristics of M. longissimus in Japanese Black heifers

The effects that high levels of protein in concentrate feed has during the early fattening stage on physico-chemical composition and sensory characteristics of M. longissimus in Japanese Black heifers were investigated. Four sets (8 head) of identical twins of Japanese Black heifers were divided into two groups: a group fed high levels of protein in concentrate feed in the early fattening stage (HCP); and a control group. Moisture, fat, protein, cooking loss, Warner-Bratzler shear force and fatty acid composition of M. longissimus were similar in both groups. Levels of free amino acid (FAA), aspartic acid, glutamine, taurine and histidine were lower in the HCP group than in the control group ( P  < 0.05). Levels of glutamic acid, serine, asparagine, threonine and total FAA in the HCP group tended to be lower than in the control group ( P  < 0.1). There were no significant differences between the sensory characteristics (juiciness, tenderness, fattiness, flavor and overall acceptability) of the two groups. These results suggest that the use of high levels of protein in concentrate feed during the early fattening stage does not affect the sensory characteristics of Japanese Black beef, but does alter the FAA content.     

Dietary supplementation with cellooligosaccharide improves growth performance in weanling pigs

Nondigestible oligosaccharides are not digested in the small intestine, but are fermented by bacteria colonizing in the large intestine. Physiological effects of non‐digestible oligosaccharides have been considered to be conferred by the fermentation of bacteria colonizing in the large intestine. Because cellooligosaccharide is a non‐digestible oligosaccharide, various physiological effects are expected. However, physiological functions of cellooligosaccharide are not well understood. This experiment was conducted to clarify the effect of dietary supplementation with cellooligosaccharide on the growth performance in weanling pigs. The result showed that average daily gain was significantly higher (P < 0.05) in pigs fed a diet supplemented with cellooligosaccharide than in pigs without cellooligosaccharide. There was a tendency to increasing average daily feed intake in pigs with cellooligosaccharide, though the significant difference was not detected (P = 0.18). Feed efficiency and nutrient digestibility of feces and ileum were not changed by feeding cellooligosaccharide. In addition, blood urea nitrogen was significantly higher (P < 0.05) in pigs fed the diet supplemented with cellooligosaccharide than in pigs without cellooligosaccharide. The concentrations of acetic and iso‐valeric acids in the cecum of pigs fed the diet with cellooligosaccharide tended to be higher (P < 0.10) than those without cellooligosaccharide. The results obtained in this study demonstrated that dietary supplementation with cellooligosaccharide improves growth performance in weanling pigs.     

Development of immune assay system for both CpG and non-CpG DNA from lactic acid bacteria using a transfectant of swine Toll-like receptor 9

Bacterial DNA is expected to be a potent immune stimulating agent to Toll‐like receptor (TLR) 9 expressed cells such as macrophages, monocytes, B lymphocytes, NK cells and dendritic cells. In the present study, we constructed a transfectant of swine TLR9 with mammalian cells. We demonstrated that the transfectant, recognizing both CpG and non‐CpG oligonucleotides from lactic acid bacteria, induced NF‐κB activation by gene reporter assay. These findings indicate that the swine TLR9 transfectant will be highly useful for the screening of immunostimulatory DNA from lactic acid bacteria.     

Physiological Specialization of Fusarium oxysporum f. sp. lactucae, a Causal Organism of Fusarium Root Rot of Crisp Head Lettuce in Japan

In 1995, Fusarium root rot of crisp head lettuce, caused by Fusarium oxysporum f. sp. lactucae, was simultaneously found in the Shiojiri and Kawakami areas of Nagano Prefecture, Japan. The Shiojiri and Kawakami isolates differed in pathogenicity to lettuce cultivars. Because of this distinct physiological specialization, these Shiojiri and Kawakami isolates should be designated as race 1 and race 2, respectively, using lines VP1010 (highly resistant to race 1), VP1013 (highly resistant to race 2) and variety Patriot (highly susceptible to both races) as differential varieties. This is the first report of races of Fusarium oxysporum f. sp. lactucae, Received 21 September 2000/ Accepted in revised form 21 March 2001     

Influence of Sulfur and Nitrogen Content of Rice Grains on Flavor in Stored Sake

The influence of nitrogen and sulfur compounds in rice grains on changes in flavor in stored sake was investigated using Japanese rice cultivars for sake making. Nitrogen content exhibited a significant positive correlation with sulfur content. Based on the molar ratio of nitrogen to sulfur in the rice grain, the sulfur compounds appeared to be derived from protein‐associated sulfur‐containing amino acids, as reported previously. The higher the protein content of the rice, the greater the amount of nitrogen and sulfur compounds found in both the digest of steamed rice grains and in the sake. Physicochemical changes were investigated in the stored sake to confirm the influence of total sulfur content. Polysulfides in the stored sake appeared to be higher when made from rice grains of high total sulfur content. Staling of stored sake was affected by levels of protein‐associated sulfur‐containing amino acids in the rice.     

Chemical characterization of milk oligosaccharides of an African lion (Panthera leo) and a clouded leopard (Neofelis nebulosa)

The Carnivora include the superfamilies Canoidea and Feloidea. In species of Canoidea other than the domestic dog, Canis lupus, the milk contains only traces of lactose and much larger concentrations of oligosaccharides. In this study, lactose was found to be the dominant saccharide in the milk or colostrum of two species of Feloidea, namely the African lion (Panthera leo) and the clouded leopard (Neofelis nebulosa). In addition to lactose, the following oligosaccharides were characterized in the milk of a lion; Neu5Gc(α2‐3)Gal(β1‐4)Glc (3′‐NGc‐SL), Fuc(α1‐2)Gal(β1‐4)Glc (2′‐fucosyllactose) and GalNAc(α1‐3)[Fuc(α1‐2)]Gal(β1‐4)Glc (A‐tetrasaccharide). The colostrum of a clouded leopard contained 3′‐NGc‐SL, Gal(α1‐3)Gal(β1‐4)Glc (isoglobotriose) and A‐tetrasaccharide. These oligosaccharides differ in some respects from those previously identified in another species of Feloidea, the spotted hyena (Crocuta crocuta). These milks contained 3′‐NGc‐SL and A‐tetrasaccharide, while spotted hyena colostrum did not; however, it contained Neu5Ac(α2‐3)Gal(β1‐4)Glc (3′‐NAc‐SL) and Gal(α1‐3)[Fuc(α1‐2)]Gal(β1‐4)Glc (B‐tetrasaccharide).