• KSBB Journal
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• v.27 no.6
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• pp.352-360
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• 2012

Bacillus clausii I-52 which produced SDS- and $H_2O_2$-tolerant extracellular alkaline protease (BCAP) was isolated from heavily polluted tidal mud flat of West Sea in Incheon, Korea and stable strain (transformant C5) of B. clausii I-52 harboring another copy of BCAP gene in the chromosome was developed using the chromosome integration vector, pHPS9-fuBCAP. When investigated the production of BCAP using B. clausii transformant C5 through pilot-scale submerged fermentation (500 L) at $37^{\circ}C$ for 30 h with an aeration rate of 1 vvm and agitation rate of 250 rpm, protease yield of approximately 105,700 U/mL was achieved using an optimized medium (soybean meal 2%, wheat flour 1%, sodium citrate 0.5%, $K_2HPO_4$ 0.4%, $Na_2HPO_4$ 0.1%, NaCl 0.4%, $MgSO_4{\cdot}7H_2O$ 0.01%, $FeSO_4{\cdot}7H_2O$ 0.05%, liquid maltose 2.5%, $Na_2CO_3$ 0.6%). The enzyme stability of BCAP was increased by addition of polyols (10%, v/v) and also, the stabilities of BCAP towards not only the thermal-induced inactivation at $50^{\circ}C$ but also the SDS and $H_2O_2$-induced inactivation at $50^{\circ}C$ were enhanced. Among the polyols examined, the best result was obtained with propylene glycol (10%, v/v). The BCAP supplemented with propylene glycol exhibited extreme stability against not only the detergent components such as ${\alpha}$-orephin sulfonate (AOS) and zeolite but also the commercial detergent preparations. The granulized enzyme of BCAP was prepared with approximately 1,310,000 U/g of granule. Wash performance analysis using EMPA test fabrics revealed that BCAP granule exhibited high efficiency for removal of protein stains in the presence of anionic surfactants as well as bleaching agents. When compared to Savinase 6T$^{(R)}$ and Everlase 6T$^{(R)}$ manufactured by Novozymes, BCAP under this study probably showed similar or higher efficiency for the removal of protein stains. These results suggest that the alkaline protease produced from B. clausii transformant C5 showing high stability against detergents and high wash performance has significant potential and a promising candidate for use as a detergent additive.

• Microbiology and Biotechnology Letters
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• v.49 no.2
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• pp.255-263
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• 2021

In this study, we assessed the technological characteristics and safety of 88 Enterococcus faecium strains isolated from meju; the strains possess the glutamate decarboxylase gene gadA/B involved in γ-aminobutyric acid production. The study was conducted to evaluate the possibility of introducing E. faecium meju isolates as food fermentation starters. We observed that a NaCl concentration of 6% (w/v) facilitated the growth and acid production of all strains. At a NaCl concentration of 7%, 21 strains (24%) exhibited a low growth rate, 72 strains (82%) a weak acid production, and 16 strains (18%) showed no acid production. All strains exhibited protease activity at a NaCl concentration of 4%. At a NaCl concentration of 5%, 86 strains exhibited weak activity, and one strain showed no protease activity. We could not detect any lipase activity in the investigated strains. None of the strains exhibited an acquired antibiotic resistance to the seven antibiotics tested in the present study, namely ampicillin, chloramphenicol, ciprofloxacin, gentamicin, penicillin G, tetracycline, and vancomycin. We could identify the Enterococcus endocarditis antigen gene efaA and the tyrosine decarboxylase gene tdc contributing to tyramine production, in 88 meju isolates. We could not detect the Enterococcus surface protein gene esp, which is specifically possessed by human-originated E. faecium strains, in any of the 88 strains tested in the study.

• Korean Journal of Poultry Science
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• v.13 no.2
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• pp.209-219
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• 1986

This study was designed to investigate the effects of various sources of dietary fats on the blood and liver cholesterol(CHOL) levels in young Single Comb White Leghorn male chicks, In experiment 1, corn oil, palm gil, tallow and fish oil were added individually at a level of 4% to semipurified type diets composed of isolated soyprotein and glucose as major components. The diets were fed ad libitum for a period of 15 days. In experiment 2, various fats such as corn oil, soybean oil, repeseed oil, palm oil, tallow, fish oil and hydrogenated fish oil(HFO) were added individually at a level of 11.4% to practical type diets primarily based on corn and soybean meal. Control diet contained 3% of corn oil. All these diets were formulated to contain equivalent amount of nutrients such as protein, vitamins and minerals on a basis of unit kcal of metabolizable energy. The third Experiment was to compare the effects of different levels of calorie/protein(C/P ratio) of diets on the performances and various biological parameters in the chicks. Control diet was the same as in experiment 2. Another diet was added with 11.14% corn oil(C/P ratio=146) and the other diet with 10% corn oil(C/P ratio=164), The diets in experiment 2 and 3 were fed ad libitum for 26 days. In the first experiment, the chicks fed the diet containing vegetable oils tended to grow faster and show better feed efficiency without significance than those fed diets added with animal fats. However, this tendency was not observed in the experiment 2. Birds consumed the diets added with fish oil appeared to have heavier liver weight and higher liver CHOL than the others(p〈0.05), No significant differences in the levels of blood CHOL and triacylglycerol(TG) were observed among the chicks of various dietary groups(Exp. 1). Weights of liver or heart were significantly heavier in the chicks consumed the diets added with HFO or fish oil, respectively(Exp. 2). However, chicks ingested diet containing fish oil appeared to have significantly lower plasma CHOL. No significant differences were observed in the levels of liver CHOL and plasma TG among the dietary groups. Birds consumed the diet with a wider C/P ratio resulted in higher liver TG levels in experiment 3(p〈0.05). Although no statistical differences were observed among the various dietary groups, chicks fed the diet with a wider C/P ratio tended to show higher levels of plasma CHOL, TG, liver CHOL and total liver lipids compared to those of the control group.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.1
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• pp.50-57
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• 2015

This study was conducted to investigate the effect of soybean meal (SM) and soluble starch (SS) on biogenic amine production and microbial diversity using in vitro ruminal fermentation. Treatments comprised of incubation of 2 g of mixture (expressed as 10 parts) containing different ratios of SM to SS as: 0:0, 10:0, 7:3, 5:5, 3:7, or 0:10. In vitro ruminal fermentation parameters were determined at 0, 12, 24, and 48 h of incubation while the biogenic amine and microbial diversity were determined at 48 h of incubation. Treatment with highest proportion of SM had higher (p<0.05) gas production than those with higher proportions of SS. Samples with higher proportion of SS resulted in lower pH than those with higher proportion of SM after 48 h of incubation. The largest change in $NH_3$-N concentration from 0 to 48 h was observed on all SM while the smallest was observed on exclusive SS. Similarly, exclusive SS had the lowest $NH_3$-N concentration among all groups after 24 h of incubation. Increasing methane ($CH_4$) concentrations were observed with time, and $CH_4$ concentrations were higher (p<0.05) with greater proportions of SM than SS. Balanced proportion of SM and SS had the highest (p<0.05) total volatile fatty acid (TVFA) while propionate was found highest in higher proportion of SS. Moreover, biogenic amine (BA) was higher (p<0.05) in samples containing greater proportions of SM. Histamines, amine index and total amines were highest in exclusive SM followed in sequence mixtures with increasing proportion of SS (and lowered proportion of SM) at 48 h of incubation. Nine dominant bands were identified by denaturing gradient gel electrophoresis (DGGE) and their identity ranged from 87% to 100% which were mostly isolated from rumen and feces. Bands R2 (uncultured bacterium clone RB-5E1) and R4 (uncultured rumen bacterium clone L7A_C10) bands were found in samples with higher proportions of SM while R3 (uncultured Firmicutes bacterium clone NI_52), R7 (Selenomonas sp. MCB2), R8 (Selenomonas ruminantium gene) and R9 (Selenomonas ruminantium strain LongY6) were found in samples with higher proportions of SS. Different feed ratios affect rumen fermentation in terms of pH, $NH_3$-N, $CH_4$, BA, volatile fatty acid and other metabolite concentrations and microbial diversity. Balanced protein and carbohydrate ratios are needed for rumen fermentation.

• Journal of Environmental Science International
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• v.23 no.6
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• pp.1095-1100
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• 2014

Keratin wastes are generated in excess of million tons per year worldwide and biodegradation of keratin by microorganisms possessing keratinase activity can be used as an alternative tool to prevent environmental pollution. For practical use of keratinase, its physicochemical properties should be investigated in detail. In this study, we investigated characteristics of keratinase produced by Xanthomonas sp. P5 which is isolated from rhizospheric soil of soybean. The level of keratinase produced by the strain P5 increased with time and reached its maximum (10.6 U/ml) at 3 days. The production of soluble protein had the same tendency as the production of keratinase. Optimal temperature and pH of keratinase were $40^{\circ}C-45^{\circ}C$ and pH 9, respectively. The enzyme showed broad temperature and pH stabilities. Thermostability profile showed that the enzyme retained 94.6%-100% of the original activity after 1 h treatment at $10^{\circ}C-40^{\circ}C$. After treatment for 1 h at pH 6-10, 89.2%-100% of the activity was remained. At pH 11, 71.6% of the original activity was retained after 1 h treatment. Although the strain P5 did not degrade human hair, it degraded duck feather and chicken feather. These results indicate that keratinase from Xanthomonas sp. P5 could be not only used to upgrade the nutritional value of feather hydrolysate but also useful in situ biodegradation of feather.

• Asian-Australasian Journal of Animal Sciences
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• v.5 no.2
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• pp.217-224
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• 1992

The study was undertaken to determine the effect of dietary magnesium on the eggshell quality and other performance in laying Tsaiya ducks and Leghorn hens. Twenty-five Tsaiya ducks and 25 Leghorn hens were raised in individual cages. The basal diet was mainly consisted of corn starch and isolated soybean protein. At the beginning of the experiments, birds were fed for 10 days with the basal diet supplemented with 500 mg/kg Mg (1070 mg/kg in total by analysis) in order to allow the birds adapting to the new diet. Both Tsaiya ducks and Leghorn hens were then each randomly divided into five groups and each group of five birds were fed with the experimental diets containing 690, 1070, 1690, 2150 or 2380 mg/kg Mg, respectively for 21 days. Eggs were collected in order to measure eggshell quality, Mg and Ca content of the eggshell. At the end of the experiments, blood samples of all birds were taken from their brachial veins for measuring the concentration of Mg and Ca in the plasma. Experimental results appeared that the dietary Mg content did not significantly affect egg production, egg weight, eggshell breaking strength and thickness in both Tsaiya ducks and Leghorn hens. In Tsaiya ducks, however, the plasma Mg concentration was affected by the dietary Mg content, but the plasma Mg almost reached a plateau (4.66 mg/dl) as long as the dietary Mg level was over 1070 mg/kg. In Leghorn hens, the plasma Mg level was significantly increased from 1.66 mg/dl to 4.03 mg/dl when the dietary Mg content in the diet had been increased from 690 mg/kg to 2380 mg/kg, suggesting that the plasma Mg concentration seems to be directly influenced by the Mg absorbed in the intestine. In the Tsaiya ducks, however, the dietary Mg level did not significantly affect the eggshell Mg content (from 0.113% to 0.123%). Whereas, there was a negative correlation between the eggshell thickness and eggshell Mg content (r = -0.50, p<0.01), revealing that the increase in eggshell Mg content probably associated with the impairment of eggshell quality in Tsaiya ducks. In Leghorn hens, however, there was no significant correlation between eggshell quality and eggshell Mg content, although the Mg content in the eggshell was also increased from 0.279% to 0.427% when the dietary Mg had been elevated from 690 mg/kg to 2380 mg/kg.

• Journal of the Korean Society of Food Science and Nutrition
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• v.22 no.2
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• pp.222-225
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• 1993

To obtain a thermal diffusivity predicting equation for meat products, minced pork meat was mixed with some additives such as lard, isolated soybean protein, 1.5% of table salt and 2% of polyphosphate to control the composition and texture of the products and then stuffed in a model can. Heat penetration curves were measured in the temperature range of 80.76~121.03$^{\circ}C$ by using a thermocouple fixed at the cold point of the model can and the thermal diffusivities were calculated from the plotted heat penetration curves. At constant heating temperature, the thermal diffusivities of pork meat with water content of 49.01~77.55% increased linearly with increasing water content. The thermal diffusivities of the products with constant water content also increased linearly with increasing heating temperature and the values could be predicted by following equation: $\alpha$p=(2.1394+0.5X$_{w}$).$\alpha$$_{w}$+0.0035.10$^{-6}$ .X$_{w}$-0.2785.10$^{-6}$ ,(m$^2$.s$^{-1}$ ). The maximal difference of the values predicted with this equation on the basis of the practical measured values were less than 1.7%. 1.7%.

• Journal of Life Science
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• v.27 no.7
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• pp.805-811
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• 2017

In the previous paper, we isolated a bacterium that can hydrolyze various organic materials from soybean paste, including cellulose, lipids, starch, and protein. The activity and chemical properties of the crude enzymes produced by the isolate Bacillus subtilis CK-2 were further investigated. Cellulase showed the highest activity at pH 5.0 and $55^{\circ}C$. The stability of cellulase was maintained within the ranges of pH 5.0~10.0 and $20{\sim}50^{\circ}C$. Cellulolytic enzymes were activated by a $Co^{2+}$ ion, demonstrating the highest activity at a 0.45%(w/v) concentration of $Co^{2+}$. The optimal conditions for amylase were pH 5.0 and $50^{\circ}C$. The activity of amylase was stable within the ranges of pH 4.0~5.0 and $20{\sim}50^{\circ}C$. The $Co^{2+}$ ion was also necessary for amylase activity, which was the highest at a 0.2%(w/v) concentration of $Co^{2+}$. The optimal pH and temperature conditions of protease were pH 8.0 and $50^{\circ}C$. The activity of protease was stable within the ranges of pH 7.0~8.5 and $20{\sim}50^{\circ}C$. Protease activity was catalyzed by $Mn^{2+}$, which was the highest at a 0.125%(w/v) concentration of $Mn^{2+}$. The isolate B. subtilis CK-2 demonstrated a high activity of autolysin. Based on these results, we identified and suggested the optimal pH, temperature, and metal ion concentration in the use of the hydrolytic enzymes of B. subtilis CK-2 for industrial purposes.