• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.10
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• pp.1535-1542
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• 2014

This study was conducted to investigate the physicochemical properties and biological activities of collagens with different molecular weights from Alaska pollack (Theragra chalcogramma) skin as well as their efficacies as functional materials. The molecular weights of collagens were between 1~10 kDa (below 1 kDa (AP1), 1~3 kDa (AP2), 3~10 kDa (AP3), and above 10 kDa (AP4). The protein content of AP4 (40.19 g/100 g) was the highest. Collagen contents of AP1, AP2, AP3, and AP4 were 36.43, 32.23, 19.23, and 14.89%, respectively. The free amino acid and essential amino acid contents of AP1 were higher than those of AP2, AP3, and AP4. Fourier transform infrared spectroscopy spectra of collagens with different molecular weights showed wavenumbers representing the regions of amide I, amide II, amide III, and amide A, respectively. The electron-donating ability (29.51%) and SOD-like activity (38.45%) of AP1 were higher than those of AP2, AP3, and AP4. Tyrosinase inhibition activity of AP1 improved with higher treatment concentration. The rate of inhibition of MMP-1 production in HS68 cells exposed to UVB was suppressed by treatment with AP1 (29.78%) and AP2 (26.49%) at 1 mg/mL. Furthermore, there was a strong correlation between DPPH, superoxide dismutase, tyrosinase activity, and MMP-1 inhibition rate of collagens with different molecular weights.

• Parasites, Hosts and Diseases
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• v.32 no.1
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• pp.35-42
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• 1994

This study was designed to evaluate the humoral immune reactions in clonorchiasis before and after praziquantel treatment. Rabbits were infected with 150 or 450 metacercariae, treated on 4 and 8111 months after infection, and observed for 13 months of posttreatment. Infection controls were maintained for 22 months. Antigen was the metabolic product of worms incubated in physiologic saline. The immune reactions of anti-clonorchis IgG were observed using SDS-PAGE/immunoblot. During the Infection and Posttreatment, the antigenic Proteins of 66, 63, 54, 52, 50,47,42, 40, 38, 34,33,30, 27, 25, 23, 20, 19, 18, 17, 16, 15, 14, 13, 12.5 and 11.5 kDa were detected. Of them, 33,27, 13, and 12.5-kDa antigens were highly antigenic and observed predominently in infection controls. After the treatment, 13 and 12.5-kDa antigens faded in 6 months after the second treatment, but 33 and 27-kDa antigens were detected until 13 months of posttreatment. The results clearly demonstrate that 13 and 12.5-kDa antigens represent attenuated host immune reactions by praziquantel treatment. As the 12.5-kDa antigen had a large amount of protein in SDS-PAGE, it was designated as'K2-Ag'of C. sinenis.

• Animal cells and systems
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• v.3 no.2
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• pp.221-228
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• 1999

In order to find a biochemical marker for late Iysosomes, we characterized two cDNAs which were cloned by using a monoclonal antibody (mAb) against Iysosomes in Amoeba proteus as a probe. The two cDNAs, a 1.3-kb cDNA in pBSK-Iys45 and a 1.6-kb cDNA in pBSK-Iys60, were found to encode proteins homologous to pepstatin-insensitive carboxyl proteinases (PICPs). E. coli transformed with pBSK-Iys45 produced two immunopositive polypeptides (45 and 43 kDa) and the cDNA in 1274 bases encoded a 44,733-Da protein (Lys45) of 420 amino acids containing one site for a core oligosaccharide. On the other hand, E. coli transformed with pBSK-Iys60 produced several polypeptides (64, 54, 45, 41, and 37 kDa) reacting with the mAb. The cDNA contained 1629 bases and encoded a 59,231-Da protein (Lys60) of 530 amino acids containing two sites for asparagine-linked core oligosaccharides. These two cDNAs showed identities of 60.3% in nucleotide sequences and 23.6% in amino acid sequences. Lys45 and Lys60 appeared to share XXEFQK as a common antigenic domain. The amino acid sequence of the Lys45 protein showed 17.4% identity and 40.9% similarity to that of PICP from Pseudomonas sp. 101. On the other hand, Lys60 showed a 24.3% identity and 51.9% similarity with human Iysosomal PICP in the amino acid sequence. A putative active center for serine protease, GTS＊xxxxxFxG, was found to be conserved among PICP homologues. The two PICPs are the first reported enzymatic markers for late Iysosomes.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.9 no.3
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• pp.800-806
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• 2008

The strain HK-12 was enriched and isolated from naturally fermented soybean for the production of fibrinolytic enzyme and the proteome of this enzyme induced during the incubation period was analyzed. The activity of fibrinolytic enzyme derived from supernatants of the HK-12 culture was performed by fibrin plate method for solid fibrinolytic activity. As the result, the fibrinolytic activity of HK-12 grown on the nutrient agar media was about 2.3 times greater than that of plasmin used as standard. The purified enzyme was prepared by a series of purification process including ammonium sulfate precipitation, DEAE-cellulose, Sephadex chromatography. The molecular weight of the enzyme was determined to approximately 23kDa with SDS-PAGE. In order to examine which strain HK-12 proteins increased or decreased during the incubation period, 2-DE analysis was performed. Protein spot #1 significantly expressed on the 2-DE gel of bacteria cultivated for 36-hrs was analysed. As the result of protein sequence analysis using MALDI-TOF MS, one protein was identified as serine protein kinase (PrkA).

• Korean Journal Plant Pathology
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• v.10 no.4
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• pp.305-313
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• 1994

Typically susceptible lesions were developed on pepper (cv. Hanbyul) leaves inoculated with the compatible strains Ds 1 of Xanthomonas campestris pv. vesicatoria. The lesions appeared first water-soaked and then turned yellow with a chlorotic area. In contrast, the leaves inoculated with the incompatible strain 81-23 initially turned yellow and then developed local necrosis. Multiplication of x. c. pv. vesicatoria in pepper leaves also were distinctly different between the two strains. The strain Ds 1 multiplied more greatly than did the strain 81-23 in the infected leaves. X. c. pv. vesicatoria infection of pepper leaves induced the synthesis of soluble proteins, especially more greatly in the compatible than in the incompatible interactions. Some pathogenesis-related (PR) proteins were detected in the intercellular washing fluid (IWF) and extracts of the infected pepper leaves. In particular, the 32 kDa protein on SDS-PAGE gels appeared intensely in the incompatible interaction. In contrast, some proteins with moluecular masses of 65, 71, and 75 kDa disappeared in the infected pepper leaves. Isoelectric focusing could identify the pIs of soluble proteins in infected pepper leaves. The accumulation of the IWF from infected leaves was more conspicuous in the incompatible than the compatible interaction. These results suggest that some extremely acidic and basic proteins were induced and accumulated in the intercellular spaces of infected pepper leaves.

• Parasites, Hosts and Diseases
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• v.35 no.1
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• pp.31-38
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• 1997

The skin ulcer in Leishmcnio mcior infection is known to be variable according to the inoculation sites even in a susceptible host. The present study traced the immunoblot patterns by the site of inoculation and duration of infection in BALB/c mice. L. mqior were subcutaneously inoculated on the nose, footpad, and back of the mice, in a dose of 3 × 106 promastigotes. Sera of the mice were collected every 10 days after inoculation. SDS-PAGE separated soluble protein bands of the promastigotes and immunoblot was carried out with the infection sera. The skin ulcer first appeared on the nose at 15 days, and on the footpad at 17 days after inoculation. The ulcer on the back appeared after 90 days. In the mice with ulcer on the nose or footpad, serum IgG antibody reacted to 202, 139, 98, 83, 81, 67, 65, 62, 59, 54, 52, 42, 26, and 23 kDa bands at 20 days after inoculation. In mice inoculated on the back, however, the immunoblot showed visible reactions with 202, 83, 81 74, 67, 65, 62, 59, 54, 52,20 and 17 kDa bands at 90 days after inoculation. The present result showed that the antigenic protein bands of L. mqior promastigotes were differed by the inoculation site and duration of infection. Since the skin ulcer and the serum antibodies to antigenic bands between 67-52 kDa appeared simultaneously, it is suggested that the serum IgG antibodies may play a role in formation of the skin ulcer in BALB/c mice.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.6
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• pp.984-991
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• 1997

New tumor suppressor gene, snm23, homologous to human nm23/NDP kinase (human nucleoside diphosphate kinase) gene whose product has a tumor metastasis inhibitory activity, was first cloned from Korean tiger shark (Scyliorhinus forazame) skin cDNA library constructed by using a $\lambda$ ZAP-II cDNA synthesis kit. About $1\times10^5$ plaques were screened and several positive plaques were isolated and confirmed by second screening. The phagemid containing a positive clone from the Uni-Zap XR vector was excised in vivo and the gene containing the tumor metastasis suppressor protein was named as snm23. Cloned gene, snm23, was sequenced with ABI-PRISM 310 Genetic Analyzer. The nucleotide and deduced amino acid sequences of snm23 have shown an open reading frame consisting of 450 base pairs that correspond to a protein of 150 amino acid residues, with a calculated molecular mass of 16.8 kDa. Sequence comparison of snm23 with human nm23/NDP kinase was performed by using Blast protein data base of National Center for Biotechnology Information. In order to determine tissue specificity, reverse transcription-polymerase chain reaction (RT-PCR) was used. Good expression level of snm23/NDP kinase was detected at the tissues from skin, cartilage, and liver of Korean tiger shark.

• Journal of Radiation Industry
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• v.4 no.1
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• pp.65-71
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• 2010

Two chitinase producing strains CHI2 and CHI4 were isolated from soybean rhizosphere soil. Both the strains belonged to Lysobacter enzymogenes as indicated by 16S rDNA sequence analysis. Though strain CHI2 and CHI4 produced extracellular chitinase, they differ in their chitinolytic activity. CHI4 produced approximately three times the higher amounts of enzyme than that of CHI2 under specified conditions. CHI2 produced $535.67U\;l^{-1}$ of chitinase after 48 h incubation with a specific activity of $3.91U\;mg^{-1}$ of protein while strain CHI4 produced $1584.13U\;l^{-1}$ of chitinase with a specific activity of $10.88U\;mg^{-1}$ protein. SDS-PAGE analysis indicated that the molecular weight of chitinase enzyme was approximately 45 kDa. A faint band with a molecular weight of 55 kDa reveals the possibility for the presence of another kind of chitin binding protein. Mutant library was developed by exposing the isolates to gamma rays at their $LD_{99}$ value (0.23 kGy). Totally, 11 mutants of CHI2 and CHI4 are reported to have enhanced chitinase activity. Several leaky mutant clones with decreased enzyme activity and a defective mutant (CHI2-M16) with complete loss of chitinase activity were also identified. CHI4-M18, CHI4-M8 and CHI4-M29 showed 78.8, 41.5, and 31.9% increased chitinase activity over wild type CHI4.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.5
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• pp.598-606
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• 2010

This study investigated the nutritional quality of soybean meal (SBM) fermented by Aspergillus ($FSBM_A$) and/or followed by Lactobacillus fermentation ($FSBM_{A+L}$). Both fermented products significantly improved protein utilization of SBM with higher trichloroacetic acid (TCA) soluble true protein content, in vitro protein digestibility and available lysine content, especially in $FSBM_{A+L}$. Moreover, $FSBM_{A+L}$ produced a huge amount of lactic acid resulting in lower pH as compared to the unfermented SBM or soybean protein concentrate (SPC) (p<0.05). $FSBM_A$ and $FSBM_{A+L}$ raised 4.14% and 9.04% of essential amino acids and 5.38% and 9.37% of non-essential amino acids content, respectively. The ${\alpha}$-galactoside linkage oligosaccharides such as raffinose and stachyose content in $FSBM_A$ and $FSBM_{A+L}$ decreased significantly. The results of soluble protein fractions and distribution showed that the ratio of small protein fractions (<16 kDa) were 42.6% and 63.5% for $FSBM_A$ and $FSBM_{A+L}$, respectively, as compared to 7.2% for SBM, where the ratio of large size fractions (>55 kDa, mainly ${\beta}$-conglycinin) decreased to 9.4%, 5.4% and increased to 38.8%, respectively. There were no significant differences in ileal protein digestibility regardless of treatment groups. SPC inclusion in the diet showed a better protein digestibility than the SBM diet. In summary, soybean meal fermented by Aspergillus, especially through the consequent Lactobacillus fermentation, could increase the nutritional value as compared with unfermented SBM and is compatible with SPC.

• Biomedical Science Letters
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• v.6 no.3
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• pp.201-208
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• 2000

The vancomycin, one of the family of glycopeptide antibiotics, inhibits the synthesis of bacterial cell wall peptidoglycan and has been widely used against gram-positive bacterial infections, especially for a treatment of methicillin resistant S. aureus infection. However, clinical isolate which was intermediately resistant to vancomycin (Mu50: MIC 8 $\mu\textrm{g}$/ml) was isolated in recent years. In this study we performed vancomycin susceptibility test with the increment method and population analysis with clinical isolates S. aureus. Also we did several kinds of tests with three selected isolates (s129: MIC 7 $\mu\textrm{g}$/ml, s134: MIC 7 $\mu\textrm{g}$/ml, s135: MIC 8 $\mu\textrm{g}$/ml) to find out possible mechanism of vancomycin resistance. As a result, the prevalence of vancomycin resistant S. aureus isolates among S. aureus strains resistant to methicillin was 23.3% (25/107). The vancomycin resistances of isolated strains of S. aureus were between those of Mu5O and Mu3 strains. By PCR analysis, none of the isolates with decreased vancomycin susceptibility contained known vancomycin resistant genes such as vanA, vanB, vanC1, vanC2, and vanH. Major bands of 81 kDa, 58 kDa, 33 kDa, 28 kDa were demonstrable in whole cell lysates by SDS-PAGE from all three isolates as well as reference strains. And especially,45 kDa protein was overproduced in Mu50 strains. Among them increased production of NAD$^{+}$-linked-$_{D}$-lactate dehydrogenase (dnLDH) were detected from one clinical strain (s135) and Mu5O strain. From these data, we suggest that the mechanism of vancomycin resistance in these isolates are distinct from that in enterococci.