• KOREAN JOURNAL OF CROP SCIENCE
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• v.47 no.2
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• pp.85-94
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• 2002

Antibodies raised against the purified p-subunit of $\beta$-conglycinin were used in immunohistochemical studies to monitor the pattern of $\beta$-conglycinin mobilization in the cotyledons during soybean [Glycine max (L.) Merr.] seed germination. Western blot analysis revealed that the break down of the $\beta$-subunit of $\beta$-conglycinin commenced as early as 2 days after seed imbibition (DAI). Concurrent with the degradation of the $\beta$-subunit of $\beta$-conglycinin, accumulation of 48, 28, and 26 kD proteolytic intermediates was observed from 2 to 6 DAI. Western blot analysis also revealed that the acidic subunit of glycinin was mobilized earlier than the basic subunit. The basic glycinin subunit was subjected to proteolysis within 2 DAI resulting in the appearance of an intermediate product approximately 2 kD smaller than the native basic glycinin subunit. In contrast to the major seed storage proteins, lipoxygenase was subjected to limited proteolysis and was detected even after 8 DAI. The first sign of $\beta$-conglycinin breakdown was observed near the vascular strands and proceeded from the vascular strands towards the epidermis. Protein A-gold localization studies using thin sections of soybean cotyledons and antibodies raised against the $\beta$-subunit of $\beta$-conglycinin revealed intense labeling over protein bodies. A pronounced decrease in the protein A-gold labeling intensity over protein bodies was observed at later stages of seed germination. The protein bodies, which were converted into a large central vacuole by 8 DAI, contained very little 7S protein as evidenced by sparse protein A-gold labeling in the vacuoles.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.5
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• pp.594-599
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• 2006

In our previous report, we indicated that not only BSA but also BGG played an important role in the allergenicity of beef. In this study, the effect of heat or high-pressure treatments to beef extract on the SDS-PAGE patterns was examined. The antigenicity of each treated samples was also investigated by Western blots assay with the sera of BGG-positive beef allergic patients. The BGG band and its antigenicity slightly disappeared but not generally in $100^{\circ}C$ group, indicating $100^{\circ}C$ treatment is not sufficient to totally eliminate the antigenicity of beef allergens. Compared with BGG band, BSA band significantly disappeared in SDS-PAGE with $100^{\circ}C$ treatment, indicating BSA is more heat- sensitive than BGG. When the beef extract was heated at $120^{\circ}C$, not only BSA but also BGG bands was largely disappeared in both SDS-PAGE and Western blots. High pressure (HP) treatment even at 600 MPa did not affect SDS-PAGE and Western blots pattern of BSA. On the contrary, BGG treated with HP showed visible changes in SDS-PAGE. 600 MPa treatment significantly reduced the antigencity. Interestingly, these behaviors of BGG were not found in the same experiments with pure BGG treated with HP. From these results, it was speculated that some kinds of proteolytic enzymes in beef extracts were involved in the BGG molecular degradation by HP treatment. The aging experiments of beef extracts treated with HP supported this hypothesis. Further studies are needed to clarify the function and working mechanism of enzymes associated with BGG degradation in beef extracts by HP treatment.

• Korean Journal of Food Science and Technology
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• v.34 no.2
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• pp.249-254
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• 2002

To determine the abilities as both lactic starter and probiotics for fermented foods, we investigated the potency of acid production, proteolytic activity and lactose metabolism of Lactobacillus amylovorus IMC-1. And the strain was cultured with lactococci in 10% skim milk medium. It was also examined the bactericidal action of antibacterial substance, produced by the strain IMC-1, against pathogenic bacteria. L. amylovorus IMC-1 showed excellent production of acid in 10% skim milk supplemented with yeast extract, and produced 0.8 and 2.7% of acid at 12 and 72 h incubation, respectively. It was found that the activity of ${\beta}-galactosidase$, about $39\;{\mu}M/minute/dry$ cell weight (mg), was stronger than that of $phospho-{\beta}-galactosidase$ in the strain IMC-1. The strain showed weak proteolytic activity in 10% skim milk, thus it produced 6 and $69\;{\mu}g/mL$ of free tyrosine at 12 and 72 h cultivation, respectively. It was known that the strain utilized mainly ${\alpha}-casein$ than ${\beta}-casein$ from patterns of SDS-PAGE. Mixed culture produced more acid than single cultures of L. amylovorus IMC-1 and Streptococcus thermophilus NIAI 510. Single culture of Str. thermophilus and mixed culture showed increasing cheese flavor with incubation times. Optimal fermentation time of mixed culture for the acid production and flora of lactic starter was 16 and 12 h by adding 0.1 and 0.5% of yeast extract to 10% skim milk, respectively. Antibacterial substance produced by the strain IMC-1 reduced about 2 log of the viable cell counts of both Escherichia coli O157 and Shigella flexneri after 24 and 4 h incubation, and they were not detected after 48 and 6 h incubation, respectively.

• The Journal of the Korean Society for Microbiology
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• v.34 no.6
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• pp.501-512
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• 1999

Gliotoxin, a fungal metabolite, is one of the epipolythiodioxopiperazine classes and has a variety of effects including immunomodulatory and apoptotic agents. This study is designed to evaluate the effect of zinc on gliotoxin-induced death of HL-60 cells. Here, we demonstrated that treatment of gliotoxin decreased cell viability in a dose and time-dependent manner. Gliotoxin-induced cell death was confirmed as apoptosis characterized by chromatin margination, fragmentation and ladder-pattern digestion of genomic DNA. Gliotoxin increased the proteolytic activities of caspase 3, 6, 8, and 9. Caspase-3 activation was further confirmed by the degradation of procaspase-3 and PARP in gliotoxin-treated HL-60 cells. Zinc compounds including $ZnCl_2$ and $ZnSO_4$ markedly inhibited gliotoxin-induced apoptosis in HL-60 cells (from 30% to 90%). Consistent with anti-apoptotic effects, zinc also suppressed the enzymatic activities of caspase-3 and -9 proteases. In addition, cleavage of both PARP and procaspase 3 in gliotoxin-treated HL-60 cells was inhibited by the addition of zinc compounds. We further demonstrated that expression of Fas ligand by gliotoxin was suppressed by zinc compounds. These data suggest that zinc may prevent gliotoxin-induced apoptosis via inhibition of Fas ligand expression as well as suppression of caspase family cysteine proteases-3 and -9 in HL-60 cells.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.6
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• pp.852-862
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• 2003

An experiment was conducted to study the effect of ionophore enriched cold processed mineral block supplemented with urea molasses on microbial growth and rumen fermentation. Twelve adult male crossbred cattle were divided into four groups on body weight basis. Animals were given wheat straw as a basal diet. The animals of group I and II were supplemented with concentrate mixture and animals of group III and IV were supplemented with cold processed urea molasses mineral block (UMMB). Thirty mg monensin/day/animal were supplemented to the animals of group II and 35 ppm monensin were incorporated in the UMMB supplemented to the animals of group IV. Dry matter (DM) intake did not differ significantly among groups. Mean rumen pH was higher in UMMB fed animals. Total volatile fatty acids (TVFA) concentration (mmole/L strained rumen liquor (SRL) in group III (113.19) was significantly (p<0.05) higher than those of group I (105.83) and II (108.74) but similar to group IV (109.34). TVFA production (mole/day) was similar in all the groups. The molar proportion of acetate was significantly (p<0.01) higher in the group I (59.56) than those of group II (51.73) and IV (55.91) but similar to group III (57.12). The molar proportion of propionate was significantly (p<0.01) higher in the monensin treated groups i.e. group II (38.38) and IV (36.26) than those of group I (27.78) and III (33.06). Butyrate molar percent was significantly (p<0.01) higher in group I (12.65) than those of group II (10.19), group III (9.83) and IV (7.84). The reduction of acetate and butyrate was due to UMMB and monensin resulted in lower A:P ratio. Average bacterial pool and bacterial production rate did not differ significantly among groups. Total N concentration (mg/100 ml SRL) was significantly (p<0.01) higher in the group I (55.30) and III (57.70) as compared to the group II (47.97) and IV (47.59). Ammonia-N concentration (mg/100 ml SRL) of group III (34.99) was significantly (p<0.01) higher than that of the group I (25.76) which was again significantly (p<0.01) higher than that of the group II (20.79) and IV (19.83) indicating slower release of ammonia due to monensin in diet. Total bacterial, cellulolytic, proteolytic bacterial and fungal count at 4 h post feeding did not differ significantly (p<0.05) among treatment groups. However, methanogenic bacterial count was significantly (p<0.01) higher in the group I (11.80) compared to the group II (8.43) which was significantly (p<0.01) higher than that of the group III (4.70) and IV (2.90). Average protozoal population was affected by both treatments. Thus feeding of UMMB and monensin in diet affected the rumen fermentation pattern towards propionate production, slower release of ammonia and reduction in methanogenic bacteria in the rumen.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.4
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• pp.587-594
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• 2010

To develop commercially available food protein hydrolysates, the effects of different types of enzymes and substrates on bitterness and solubility of partially hydrolyzed food proteins were investigated. Four types of proteins (casein, isolated soy protein (ISP), wheat gluten, and gelatin) and five types of proteolytic enzymes (a microbial alkaline protease (alcalase), a microbial neutral protease (neutrase), papain, bromelain, trypsin) were used. To profile the pattern of hydrolysis, the degree of hydrolysis (DH) were monitored during 180 min of reaction time by pH-stat method. Casein showed the highest susceptibility to hydrolysis for all five proteases compared to those of ISP, gluten, and gelatin. In addition, the bitter intensity and solubility (nitrogen soluble index, NSI) of each protein hydrolysate were compared at DH 10%. Bitterness and solubility of protein hydrolysates were highly affected by DH and the types of enzymes and substrates. At DH=10%, casein hydrolysate by trypsin, ISP and gluten hydrolysates by either bromelain or neutrase, and gelatin hydrolysates by the five proteases tested in this study were highly soluble and less bitter.

• Korean Journal of Food Science and Technology
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• v.25 no.1
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• pp.39-45
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• 1993

The effects of enzymatic modification with pepsin and actinidin was studied on molecular weight distributions and functional properties of hydrolysates from soy protein isolate (SPI) differing in degree of hydrolysis. The hydrolyzed SPI by pepsin showed 41.5% degree of hydrolysis after 5 min, and maximum hydrolysis was obtained after 2 hours. Actinidin hydrolyzed SPI 26.71% degree after 1 hour. On SDS-PAGE, native SPI showed 9 distinguishable bands on SDS-PAGE gel. Pepsin treated SPI showed one broad band in the lower part of gel. This band was shifted further to the bottom of the gel and became faint as hydrolysis time increased. While actinidin treated SPI showed different SDS-PAGE pattern from pepsin. However PAGE patterns were similar with pepsin and actinidin treated groups. With pepsin treatment, solubility of SPI distinctively increased around isoelectric point(pI). Emulsifying activity (EA) and emulsifying stability (ES) showed marked increase over pH range of $3.0{\sim}8.0$. 5 min modified group had most excellent foam expansion (FE). Foam stability (FS) was increased as pepsin treatment time increased at pI. With actinidin treatment, solubility was increased. 60 min modified SPI had the most effective EA at pH 4.5. However ES was not effected by actinidin treatment. 5 min modified group was most effect in FE. FS was higher at alkaline pH.

• Clinical and Experimental Reproductive Medicine
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• v.10 no.1
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• pp.7-24
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• 1983

On the basis of the semen analysis in 66 subjects, they were divided into six different groups: Group I consisted of 16 normal subjects with sperm counts of over 40 ${\times}10^6$/ml and motility of over 40 percent, Group II, 7 subjects with normal sperm counts, but motility of under 40 percent, Group III, 15 oligospermic patients with under 40 ${\times}10^6$/ml, Group IV 14 azoospermic patients, Group V, 10 patients with vasectomy and Group VI, 4 abnormal patients with 2 cases of hypoplastic testis, 1 case of Klinefelter's syndrome and 1 case of testis tumor. After seperation of semen into sperm and seminal plasma by centrifugation, the protein contents and the activities of hyaluronidase, ${\beta}$-N acetylglucosaminidase, ${\beta}$-glucuronidase, arylsulfatase, acrosin and azocoll proteinase in seminal plasma were measured. Vasectomy group has 30 percent less of total protein than normal group. For the comparison of enzyme activities of seminal plasma, it could be assumed that the enzymes in seminal plasma were not contaminated with the enzymes of spermatozoa by testing the enzymes of the seminal plasma from the vasectomy and azoospermic groups. It had been reported that hyaluronidase was only released from spermatozoa, however, the result obtained in this investigation showed that azoospermic and vasectomy group had high specific activities of hyaluronidase. The results indicated that hyaluronidase was not only from the testis but also from the male accessory sexual glands. Oligospermic group (Group III) showed the lowest total activity of hyaluronidase among them. The specific activities of ${\beta}$ -N-acetylglucosaminidase was high in oligospermic group (Group III) and low in vasectomy group (Group V). These results were contradictory with the pattern of hyaluronidase activities. This indicated that the spermatozoa which were stayed in epididymis would increase the activity of this enzyme. The specific activity of ${\beta}$ - glucuronidase was low in oligospermic and vasectomy groups. Group VI including testis tumor had remarkably high arylsulfatase activity. Arylsulfatase, a typical lysosomal enzyme, has been known to be released unusually large amounts from certain tumor cells. Arylsulfatase was also released with high activities from azoospermic and vascetomy group. This result indicated that this enzyme was also released from the sources other than testis. Acrosin, a proteolytic enzyme locating in the sperm acrosome, was not found throughout all the samples of seminal plasma. The activities of azocoll proteinase, a non-specific neutral proteinase was nearly identical in all the groups. This enzyme must have been released from the sources other than testis.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.11
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• pp.1653-1658
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• 2002

The objectives of this study were to examine calpain activity and meat tenderness by three different feeding patterns in Korean native cattle (KNC). Total forty-five animals were assigned each fifteen in long term restriction feeding (LTFR), long-term restriction feeding and hormone treatment (LTFR-tH), and short term non-restriction feeding (STFNR), respectively. Concentrate was restricted based on body weight in exp 1 and 2. However, it was fed ad libitum in exp. 3. Hormonal implantation was made with $M-PO^{TM}$ for bulls and with $F-TO^{TM}$ for heifers at 18, 20, 22 months of age in exp. 2. Animals were purchased (3-5 month old) from local cattle market and managed in two local farms and university research unit at three different years. Animals were slaughtered at 24 months for long-term trial and at 18 month for short term-trial. Loin and tender loin muscle was used for calpain activity and meat quality. Calpain proteolytic system was not changed by treatment. However, calpastatin activity was low in short-term trial. The calpain and calpastatin activity is reciprocal relationship, therefore, the high calpain activity may effect on quality grade. The shear force value was decreased as the processing of aging after postmortem. On the other hand, the cooking loss was significantly higher in short-term than in long-term trial, and then gradually decreased by the aging. Hormone implants to increase meat yield influenced to calpastatin activity more powerfully than calpain activity to meat tenderness. In meat color-a*, there was not significant difference in loin. Meat color-b* was decreased as postmortem aging time increased in tenderloin. Western blots were done to learn whether these proteins are degraded during postmortem storage and whether this degradation temporally parallels the decrease of shear force value. Vinculin was detected at 0 day and 1 day and degraded after 3 day. In conclusion, Calpain activity was affected slightly on meat tenderness. But meat tenderness was influenced by calpastatin, more effectively.

• Food Science and Preservation
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• v.13 no.1
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• pp.71-76
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• 2006

We monitored the characteristics of soybean hydrolysate prepared under various hydrolysis condition using response surface methodology. The yield was affected by protease content but 1be effect of hydrolysis time to yield gradually increased at over $0.4\%$ of protease, while the $R^2$ of polynomial equation was 0.978 (p<0.01). The soluble solid enlarged by increase of both variables and the $R^2$ of polynomial equation was 0.954 (p<0.01). The degree of hydrolysis was affected by protease content at low (under $0.4\%$) protease and maximized at $0.57\%$ protease and 5.49 hrs. The $R^2$ of polynomial equation for the degree of hydrolysis was 0.916 (P<0.05). The calcium intolerance capacity showed similar pattern like yield but the effect of hydrolysis time was rapidly increased at over $0.4\%$ protease. The $R^2$ of polynomial equation for calcium intolerance capacity was 0.932 (p<0.05). The total phenolic compounds increased in proportion to protease content and hydrolysis time, while the $R^2$ of polynomial equation was 0.920 (p<0.05). According to the results of this study, the optimal conditions for soybean hydrolysis were predicted to be $0.51\~0.66\%$ of protease and $6.5\~9.0\;hrs$, and the predicted values and actual values of each response variable were similar to each other when the hydrolysis was performed at a random point within the optimal range.