• Korean journal of applied entomology
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• v.47 no.4
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• pp.457-465
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• 2008

Eight Bacillus thuringiensis strains activated against mosquito larva were compared their characterization. Spherical-shaped parasporal inclusion of B. thuringiensis subsp. tohokuensis CAB167 was observed by phase-contrast microscopy and scanning electron microscopy. $LC_{50}$ values of B. thuringiensis subsp. tohokuensis CAB167 against Culex pipiens molestus, Culex pipiens pallens, and Aedes aegyti were 173, 190 and 580 ng/ml, respectively. B. thuringiensis subsp. tohokuensis CAB167 had a parasporal inclusion containing 4 major protein components, for example, 135, 80, 49 and 28-kDa by SDS-PAGE. Otherwise, after trypsin digestion of parasporal inclusion, SDS-PAGE was showed new protease-resistant peptides at 72 and 63-kDa. Activated toxins of isolated CAB167 were different from other reference strains on a serological by immuno-diffusion test.

• Korean Journal of Agricultural Science
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• v.44 no.4
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• pp.513-518
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• 2017

Soybean, one of most widely used swine feed component in the world, contains non-starch polysaccharides (NSP). The digestive system of weaned pigs is not yet fully developed, and thus weaned pigs cannot easily digest diets based on corn and soybean meal. Dietary exogenous enzymes supplementation has been intensively investigated to assist digestion of anti-nutritional factors, such as NSP. This experiment was conducted to investigate the effects of dietary enzyme cocktail on growth performance, intestinal morphology, and nutrient digestibility of weaned pigs. A total 36 weaned pigs ($5.92{\pm}0.48kg\;BW$; 28 d old) were randomly allotted to 2 dietary treatments (3 pigs/pen, 6 replicates/treatment) in a randomized complete block design. The dietary treatments were a typical diet based on corn and soybean meal (CON) and CON with 0.05% enzyme cocktail (Cocktail; mixture of xylanase, ${\alpha}-amylase$, protease, ${\beta}-glucanase$, and pectinase). Pigs were fed their respective diets for 6 wk. Growth performance, morphology of ileum, apparent ileal digestibility (AID) and apparent total tract digestibility (ATTD) of dry matter, crude protein, and energy of weaned pigs were measured. No significant differences (p > 0.05) were observed for growth performance for the duration of the experimental period, and morphology of ileum, and nutrient digestibility between CON and Cocktail treatment groups. Therefore, the results from the current study indicated that enzyme cocktail supplementation in diets had no influence on growth performance, intestinal morphology, and nutrient digestibility of weaned pigs.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.4
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• pp.544-550
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• 2015

This study was conducted to investigate effects of heat-oxidized soy protein isolate (HSPI) on growth performance, serum biochemical indices, apparent nutrient digestibility and digestive function of broiler chickens. A total of 320 1-day-old Arbor Acres chicks were randomly divided into 4 groups with 8 replicates of 10 birds, receiving diets containing soy protein isolate (SPI, control group) or the same amount of SPI heated in an oven at $100^{\circ}C$ for 1, 4, or 8 h, for 21 days, respectively. The results indicated that compared with the control group, body weight gain and feed intake of birds fed diet containing SPI heated for 8 h were significantly lower (p<0.05). Serum urea nitrogen concentration was higher in the broilers fed diet containing SPI heated for 4 or 8 h at d 21 (p<0.05). In contrast, serum glucose content was decreased by HSPI substitution at d 21 (p<0.05). The relative pancreas weight in HSPI groups was higher than that in the control group at d 21 (p<0.05). Meanwhile, the opposite effect was observed for relative weight of anterior intestine and ileum in broilers fed a diet containing SPI heated for 8 h (p<0.05). Birds fed diets containing SPI heated for 4 or 8 h had a decreased lipase activity in anterior intestinal content at d 14 and 21 (p<0.05), respectively. In addition, the same effect was also noted in broilers given diets containing SPI heated for 1 h at d 21 (p<0.05). Similarly, amylase, protease and trypsin activity in anterior intestinal content were lower in broilers fed diets containing SPI heated for 8 h at d 21 (p<0.05). The apparent digestibility of dry matter (DM) from d 8 to 10 and DM, crude protein (CP), and ether extract from d 15 to 17 were lower in broilers fed diets containing SPI heated for 8 h (p<0.05). Besides, birds given diets containing SPI heated for 4 h also exhibited lower CP apparent digestibility from d 15 to 17 (p<0.05). It was concluded that HSPI inclusion can exert a negative influence on the growth performance of broilers, which was likely to result from the simultaneously compromised digestive function.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2001.06a
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• pp.40-45
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• 2001

Raw starch-digesting amylase (BF-2A, M.W. 93, 000 Da) from Bacillus circulans F-2 was converted to two components during digestion with subtilisin. Two components were separated and designated as BF-2A' (63, 000 Da) and BF-2B (30, 000 Da), respectively. BF-2A' exhibited the same hydrolysis curve for soluble starch as the original amylase (BF-2A). Moreover, the catalytic activities of original and modified enzymes were indistinguishable in $K_{m}$, Vmax for, and in their specific activity for soluble starch hydrolysis. However, its adsorbability and digestibility on raw starch was greatly decreased. Furthermore, the enzymatic action pattern on soluble starch was greatly different from that of the BF-2A. A smaller peptide (BF-2B) showed adsorb ability onto raw starch. By these results, it is suggested that the larger peptide (BF-2A') has a region responsible for the expression of the enzyme activity to hydrolyze soluble substrate, and the smaller peptide (BF-2B) plays a role on raw starch adsorption. A similar phenomenon is observed during limited proteinase K, thermolysin, and endopeptidase Glu-C proteolysis of the enzyme. Fragments resulting from proteolysis were characterized by immunoblotting with anti-RSDA. The proteolytic patterns resulting from proteinase K and subtilisin were the same, producing 63- and 30-kDa fragments. Similar patterns were obtained with endopeptidase Glu-C or thermolysin. All proteolytic digests contained a common, major 63-kDa fragment. Inactivation of RSDA activity results from splitting off the C-terminal domain. Hence, it seems probable that the protease sensitive locus is in a hinge region susceptible to cleavage. Extracellular enzymes immunoreactive toward anti-RSDA were detected through whole bacterial cultivation. Proteins of sizes 93-, 75-, 63-, 55-, 38-, and 31-kDa were immunologically identical to RSDA. Of these, the 75-kDa and 63-kDa proteins correspond to the major products of proteolysis with Glu-C and thermolysin. These results postulated that enzyme heterogeneity of the raw starch-hydrolysis system might arise from the endogeneous proteolytic activity of the bacterium. Truncated forms of rsda, in which the gene sequence encoding the conserved domain had been deleted, directed the synthesis of a functional amylase that did not bind to raw starch. This indicates that the conserved region of RSDA constitutes a raw starch-binding domain, which is distinct from the active centre. The possible role of this substrate-binding region is discussed.d.

• Korean Journal of Food Science and Technology
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• v.34 no.5
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• pp.867-872
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• 2002

This study was performed to investigate the optimum process conditions for manufacturing yeast extract from waste brewer's yeast using various enzymes. Contents of IMP, GMP, free amino acids, and crude protein of yeast extracts were measured by enzymes treatment. Crude protein contents of yeast extracts subjected to cell wall digestion enzyme treatment were 21.1, 33.6, and 28.0% for the control grouup, glucanase (0.5%, 12 h), and tunicase (1%, 18 h), respectively. Crude protein contents of yeast extracts subjected to protease treatment were 22.0, 30.8, and 29.8% for control group, bromelin (1%, 3 h), and protamex (1%, 3 h), respectively. Crude protein content of yeast extract subjected to glucanase and protamex mixed treatment was 34.4%. The total contents of IMP and GMP of yeast extracts subjected to G+P+A (glucanase+phosphodiesterase+adenyldeminase) and G+Pro+P+A (glucanase+protamex+phosphodiesterase+adenyldeaminase) treatments were 1,066 and 1,047 mg/100 g, respectively. The content of free amino acids of yeast extract was the highest (2,302 mg/100 g) in G+Pro+P+A treatment. Optimum concentration and process condition of enzyme treatment to obtain yeast extract with high IMP, GMP, and free amino acid content were in the order of glucanase (0.5%, 12 h), protamex (1%, 3h), phosphodiesterase (0.1%, 3 h) and adenyldeaminase (1%, 1.5 h) treatments.

• Journal of Dairy Science and Biotechnology
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• v.21 no.2
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• pp.97-104
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• 2003

It is extremely important to destroy the antigenicity of milk proteins for dietetic treatment of infants with milk allergy. Enzymatic digestion of milk protein is not only effective for destroying antigenicity, but it also is less liable to alter the nutritive value. Bovine casein was hydrolyzed with eight different commercial proteases derived from bacterias or fungi, either individually or in combination to eliminate protein allergenicity. The average molecular weight of casein hyrdolysates determined by size exclusion chromatography is about 550${\sim}$2,300 dalton range. Antigenicity of the casein hyrdolysates was not detected by heterologous passive cutaneous anaphylaxis in guinea pig-rabbit antiserum system. The inhibition test on the enzyme-linked immunosorbent assay(ELISA) showed that the antigenicity of casein hydrolysates is lowed up to 1/8,000 than that of intact bovine casein. As the enzyme reaction was carried out by the combination of bacterial and fungal protease, casein hydrolysates showed much lower bitterness and antigenicity. It suggests that these hydrolysates will be applied to many kinds of foods including the development of hypo-allergenic infant formula.

• Food Science of Animal Resources
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• v.29 no.6
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• pp.659-667
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• 2009

Casein is considered to be the main source of protein in milk; therefore, many studies have been conducted to identify casein-derived bioactive peptides and their physiological effects. Casein is inactive within the parent protein but can be liberated by various proteases and enzymatic hydrolysis during microbial fermentation and gastrointestinal digestion. Once absorbed, casein exhibits different bioavailabilities in the body. Specifically, casein-derived peptides function as angiotensin converting enzyme (ACE) inhibitor in the cardiovascular system; thus, they are expected to reduce and prevent hypertension. Additionally, casein-derived peptides behave as opioid-like peptides in the nervous system, which impacts relaxation. These peptides are also expected to modulate various aspects of immune functions. Finally, caseinophosphopeptide (CPP) and glycomacropeptide (GMP) may exhibit a number of nutritional effects such as the absorption of calcium, iron or zinc. Many studies have been conducted to evaluate casein-derived peptides due to their multifunctional properties and the results of these studies have contributed to the development of a wide variety of functional dairy products. The purpose of this paper was to review the generation of bioactive peptides, their absorption and metabolism, and their specific bioactive effects.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.45 no.4
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• pp.389-397
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• 2019

Lysosomes are cellular organelles involved in energy metabolism and intracellular digestion in eukaryotic cells, including protease, nuclease, glycosidase, lipase, and phosphatase. Our previous studies have confirmed that egg white lysosomes had melanin decolorization and reduction activity. However, there have been few studies on skin barrier and skin regeneration as well as inhibition of melanin production by egg white lysosomes on B16F10 melanocyte cell line. In this study, we attempted to identify the effect of lysosome-related organelle extract (LOE) extracted from egg white on the melanin content change and skin barrier enhancement in cells. First, cytotoxicity evaluation was performed on B16F10 melanocyte cell line to confirm the whitening efficacy of LOE. Cytotoxicity by LOE was not observed at 20 mg/mL concentration, but cytotoxicity was observed at 40 mg/mL, and the maximum concentration value was set to 20 mg/mL in all subsequent experiments. LOE samples of 5, 10, 20 mg/mL inhibited melanin production by 61.5 ± 4.0%, 61.4 ± 7.3%, 58.3 ± 8.3%, respectivly, compared to α-MSH, a negative control in melanin contents assay. MITF mRNA expression was reduced by about 39.7 ± 3.2% compared to the α-MSH treatment group. TEER assay using HaCaT showed that LOE increased TEER resistance in a dose-dependent manner, indicating that LOE is involved in strengthening the skin barrier. LOE also increased the TEER resistance under TNF-α treatment. Skin barrier was normally restored by LOE even under the condition of inflammation. LOE had a positive effect on cell division and cell migration promotion, confirmed by the observing the effect of promoting cell migration by LOE through cell migration assay. Taken together, we expect that LOE can be developed as a cosmetic material to enhance has effects on skin regeneration and skin barrier strengthening as well as whitening function if enzyme stabilization and formulation technology are combined.

• Radiation Oncology Journal
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• v.19 no.2
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• pp.153-162
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• 2001

Purpose : The mechanical insights of death of cancer cells by ionizing radiation are not of yet clearly defined. Recent evidences have demonstrated that radiation therapy may induce cell death via activation of signaling pathway for apoptosis in target cells. This study is designed whether ionizing radiation may activate the signaling cascades of apoptosis including caspase family cysteine pretenses, $Bcl_2/Bax$, cytochrome c and Fas/Fas-L in target cells. Materials and Methods : HL-60 cells were irradiated in vitro with 6 MV X-ray at dose ranges from 2 Gy to 32 Gy. The cell viability was tested by M assay and the extent of apoptosis was determined using agarose gel electrophoresis. The activities of caspase proteases were measured by proteolytic cleavages of substrates. Western blot analysis was used to monitor PARP, Caspase-3, Cytochrome-c, Bcl-2, Bax, Fas and Fas-L. Results : Ionizing radiation decreases the viability of HL-60 cells in a time and dose dependent manner. Ionizing radiation-induced death in HL-60 cells is an apoptotic death which is revealed as characteristic ladder-pattern fragmentation of genomic DNA over 16 Gy at 4 hours. ionizing radiation induces the activation of caspase-2, 3, 6, 8 and 9 of HL-60 cells in a time-dependent manner. The activation of caspase-3 pretense is also evidenced by the digestion of poly (ADP-ribose) polymerase and procaspase 3 with 16Gy ionizing irradiation. Anti-apoptotic Bcl2 expression is decreased but apoptotic Bax expression is increased with mitochondrial cytochrome c release in a time- dependent manner. In addiiton, expression of Fas and Fas-L is also increased in a time dependent manner. Conclusion : These data suggest that ionizing radiation-induced apoptosis is mediated by the activation of various signaling pathways including caspase family cysteine proteases, $Bcl_2/Bax$, Fas and Fas-L in a time and dose dependent manner.

• The Korean Journal of Mycology
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• v.40 no.4
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• pp.229-234
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• 2012

A key molecule in the pathogenesis of Alzheimer's disease (AD) is the ${\beta}$-amyloid peptide ($A{\beta}$) generated by ${\beta}$-secretase activity, an aspartic protease. This study was designed to evaluate inhibitory effect of the high-molecular weight water-soluble polysaccharides (Et-P) isolated and purified from Phellinus linteus fruiting body on ${\beta}$-secretase activity. The Et-P was purified from the hot water extract of Phellinus linteus fruiting body mainly by 75% ethanol precipitation and DEAE-Cellulose column chromatography. From the DEAE-Cellulose chromato-gram and molecular weight analysis, the Et-P was shown to be a mixture of three polysaccharides with molecular mass of 1,629, 1,294, and 21 kDa, respectively. The monosaccharide composition of Et-P was determined to be glu-cose, galactose, and mannose as major sugars, glucose being the most prominent one (48% in mole percentage). The elemental analysis and FT-IR analysis suggested that Et-P is typical polysaccharides having at least partially ${\beta}$-linkages and possible existing as complex with phenolic compounds. The laminarinase digestion and HPAEC-PAD analysis suggested that Et-P is a variant of beta-(1,3)-glucans. The Et-P showed DPPH radical scavenging activity and, especially, a significant inhibitory activity on ${\beta}$-secreatase activity (48% inhibitin at 100 ${\mu}g/mL$), suggesting that they may inhibit the formation of $A{\beta}$ which is the major causative of Alzheimer's disease. The results of this study suggest that the water soluble polysaccharides of Phellinus linteus fruiting body can be a potent material for the development of preventive or therapeutic agents for AD.