Jun-Sang Ham

Complete Genome Sequence of Bifidobacterium longum subsp. longum KACC 91563

Bifidobacterium longum strains predominate in the colonic microbiota of breast-fed infants. Here we report the complete genome sequence of B. longum subsp. longum KACC 91563, isolated from feces of neonates. A single circular chromosome of 2,385,301 bp contains 1,980 protein-coding genes, 56 tRNA genes, and 3 rRNA operons.

Genome Sequence of Lactobacillus mucosae LM1, Isolated from Piglet Feces

Lactobacillus mucosae LM1, isolated from stool samples of a healthy piglet, displays good in vitro mucin adhesion and antimicrobial activity against pathogenic bacteria. To elucidate its antimicrobial effects and to find its epithelial cell and mucin adhesion genes, the genomic sequence of L. mucosae LM1 was investigated.

Extracellular vesicle-derived protein from Bifidobacterium longum alleviates food allergy through mast cell suppression.

BACKGROUND The incidence of food allergies has increased dramatically during the last decade. Recently, probiotics have been studied for the prevention and treatment of allergic disease. OBJECTIVE We examined whether Bifidobacterium longum KACC 91563 and Enterococcus faecalis KACC 91532 have the capacity to suppress food allergies. METHODS B longum KACC 91563 and E faecalis KACC 91532 were administered to BALB/c wild-type mice, in which food allergy was induced by using ovalbumin and alum. Food allergy symptoms and various immune responses were assessed. RESULTS B longum KACC 91563, but not E faecalis KACC 91532, alleviated food allergy symptoms. Extracellular vesicles of B longum KACC 91563 bound specifically to mast cells and induced apoptosis without affecting T-cell immune responses. Furthermore, injection of family 5 extracellular solute-binding protein, a main component of extracellular vesicles, into mice markedly reduced the occurrence of diarrhea in a mouse food allergy model. CONCLUSION B longum KACC 91563 induces apoptosis of mast cells specifically and alleviates food allergy symptoms. Accordingly, B longum KACC 91563 and family 5 extracellular solute-binding protein exhibit potential as therapeutic approaches for food allergies.

Genome Sequence of Lactobacillus salivarius GJ-24, a Probiotic Strain Isolated from Healthy Adult Intestine

The draft genome sequence of Lactobacillus salivarius GJ-24 isolated from the feces of healthy adults was determined. Its properties, including milk fermentation activity and bacteriocin production, suggest its potential uses as a probiotic lactic acid bacterium and start culture for dairy products.

Genome Sequence of Lactobacillus johnsonii PF01, Isolated from Piglet Feces

Lactobacillus johnsonii PF01, an autochthonous bacterium of the gastrointestinal tract, was isolated from a fecal sample from a piglet. The strain adhered specifically to the duodenal and jejunal epithelial cells of the piglet and had high bile resistance activity. Here we report the genomic sequence of L. johnsonii PF01.

Identification of Antihypertensive Peptides Derived from Low Molecular Weight Casein Hydrolysates Generated during Fermentation by Bifidobacterium longum KACC 91563

Angiotensin-converting enzyme (ACE) inhibitory activity was evaluated for the low-molecular-weight fraction (<3 kDa) obtained from milk fermentation by Bifidobacterium longum KACC91563. The ACE inhibitory activity in this fraction was 62.3%. The peptides generated from the <3 kDa fraction were identified by liquid chromatography-electrospray ionization quantitative time-of-flight mass spectrometry analysis. Of the 28 peptides identified, 11 and 16 were identified as β-casein (CN) and αs1-CN, respectively. One peptide was identified as κ-CN. Three peptides, YQEPVLGPVRGPFPIIV, QEPVLGPVRGPFPIIV, and GPVRGPFPIIV, from β-CN corresponded to known antihypertensive peptides. We also found 15 peptides that were identified as potential antihypertensive peptides because they included a known antihypertensive peptide fragment. These peptides were as follows: RELEELNVPGEIVE (f1-14), YQEPVLGPVRGPFP (f193-206), EPVLGPVRGPFPIIV (f195-206), PVLGPVRGPFPIIV (f196-206), VLGPVRGPFPIIV (f197-206), and LGPVRGPFPIIV (f198-206) for β-CN; and APSFSDIPNPIGSENSEKTTMPLW (f176-199), SFSDIPNPIGSENSEKT- TMPLW (f178-199), FSDIPNPIGSENSEKTTMPLW (f179-199), SDIPNPIGSENSEKTTMPLW (f180-199), DIPNPIGSENSEKTTMPLW (f181-199), IPNPIGSENSEKTTMPLW (f182-199), PIGSENSEKTTMPLW (f185-199), IGSENSEKTTMPLW (f186-199), and SENSEKTTMPLW (f188-199) for αs1-CN. From these results, B. longum could be used as a starter culture in combination with other lactic acid bacteria in the dairy industry, and/or these peptides could be used in functional food manufacturing as additives for the development of a product with beneficial effects for human health.

Isolation and Characterization of Cellulase Secreting Bacterium from Cattle Manure: Application to Composting

A extracellular cellulase producing bacterium was isolated from cow feces, and was identified to be Bacillus licheniformis on the basis of morphological and biochemical properties as well as the composition of cellular fatty acids composition (FAME). CMCase, FPase and avicelase activities of the isolates cultured in CMC media at 37°C for 24 hrs were 1.65 U/ml, 0.13 U/ml and 0.18 U/ml, respectively. However, β-glucosidase activity was not detected. The optimum pH and temperature for induction of the enzyme were 7.5 and 50°C. The maximum CMCase activity was observed at pH 7.5 and 75°C. Zymogram analysis for crude supernatant showed four major bands CMC-SDS-PAGE. By adding 0.1% DMP (Developed Microbial Product) -comp including B. licheniformis NLRI X-33, microbial product, to swine manure composting pile, the gab of composting temperature within the pile in DMP-comp and control was 23°C and 34°C, respectively, during the initial 23 day composting periods. Coliforms were not found in the swine + sawdust samples after 30 days of DMP-comp treated and 40 days of control under forced aeration system.

Effect of Dietary Supplementation of Quercetin on Oxidative Stability of Chicken Thigh

This study was carried out to determine the effect of dietary supplementation of quercetin and methoxylated quercetin extracted from onions on oxidation of chicken thigh during cold storage. For 35 days, 1-day-old 320 broiler chicks (Ross) were divided into 8 groups and supplemented the diet; basal diet only (T1), T1 with antibiotics (T2), vitamin E 20 IU (T3), vitamin E 200 IU (T4), quercetin 20 ppm (T5), quercetin 200 ppm (T6), methoxylated quercetin 20 ppm (T7), methoxylated quercetin 200 ppm (T8). After slaughtering the broilers, thigh meats were collected and analyzed the oxidation stability during storage at 4 for 7 days. Volatile basic nitrogen (VBN) was ranged from 10.98 15.37 mg% until day 3 and quercetin 20 ppm (T5) significantly reduced (p<0.05) the VBN value of thigh as much as antibiotics treatment group (T2). The supplementation of quercetin at a 200 ppm level significantly reduced 2-thiobarbituric acid reactive substances value when compared with the control. Supplementation of 20 ppm quercetin also affected fatty acid composition of chicken thigh and resulted in increase of C18:1w9 and decrease of C18:2w6 on day 3. In microbes of chicken thigh during storage, supplementation of 20 ppm quercetin significantly reduced total microbes compare to control on day 7 (p<0.05). There was no effect on the number of coliforms in thigh with quercetin diet. From these results, the dietary supple- mentation of 20 ppm of quercetin may have effects on enhancing oxidation stability.

Development of Ice Cream with Improved Microbiological Safety and Acceptable Organoleptic Quality Using Irradiation

To develop the manufacturing method of ice cream with microbiologically safe and proper sensory quality using irradiation for sensitive consumer, 3 different flavors, which were resistant to their flavors against irradiation, were selected and used for ice cream manufacturing to reduce the irradiation-induced off-flavor problem. The general composition was not different among treatments. Total aerobic bacteria were detected as 2.38, 1.23, 1.38, and 1.15 log CFU/g level in ice cream with control(no flavor added), spearmint, mint, and citrus flavor, respectively. No viable cells were observed by irradiation at 1 kGy except for the control. Sensory evaluation indicated that the irradiated ice cream with spearmint flavor at 1 kGy and citrus flavor at 3 kGy had higher overall acceptability. Therefore, a low dose irradiation (less than 3 kGy) with mint or citrus flavors may enhance the safety of ice cream with proper sensory quality for sensitive consumer.

Determination of molecular weights of caprine milk proteins by matrix-assisted laser desorption/ionization mass spectrometry

Molecular weights (MW) of major proteins in milk of 3 Korean dairy goat breeds were determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, after treatment of milk samples with the reduction buffer used in capillary electrophoresis. The MW of caprine milk proteins were compared with those of Holstein milk counterparts using commercial bovine milk protein standards. The MW of α-lactalbumin, β-lactoglobulin, and α- and β-casein standards were 14,197±3.4, 18,326±26.3, 23,591±13.0, and 23,967±12.8 m/z, respectively, whereas those of Holstein milk treated with the reduction buffer were 14,199±8.3, 18,397±25.9, 23,614±64.8, and 23,984±75.6 m/z, respectively. The respective MW of α-lactalbumin in Saanen, Toggenberg, and Alpine milk were 14,194±27.2, 14,266±105.9, and 14,241±13.2 m/z, which were not different from those of the bovine milk. The respective MW of β- lactoglobulin in corresponding caprine milk were 18,840±31.5, 18,856±26.3, and 18,857±21.3 m/z, which were higher than those in the bovine milk. The MW of β-casein in corresponding caprine milk were 23,860±27.2, 23,886±12.3, and 23,901±8.4 m/z, which were lower than those in the bovine milk. The results indicated that matrix-assisted laser desorption/ionization time-of-flight mass spectrometry could be used for rapid determination of MW of Korean caprine milk proteins without protein separation steps.