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Physicochemical and Functional Properties of Pepsin-modified Myofibrillar protein from Sardine, Sardinops melanostica (Pepsin으로 수식된 정어리 myofibrillar protein의 특성)

  • Kim, Byung-Mook;Kim, Byung-Ryul
    • Korean Journal of Food Science and Technology
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    • v.26 no.2
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    • pp.110-116
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    • 1994
  • In order to study the effects of enzyme modification on the physico-chemical and functional properties of myofibrillar protein prepared from the frozen sardine, Sardinops melanostica, the protein was hydrolyzed with pepsin under the enzyme-substrate ratio 1:100 at $37^{\circ}C$ and pH 1.65 for 1, 4, 8, 12, and 24 hr, respectively. The properties of pepsin-modified sardine myofibriliar protein were determined. The extents of proteolysis with pepsin as a fuction of time was showed a typical enzyme hydorlysis curve with an initial region of 4 hour period followed by plateau region. The SDS-acrylamide slab gel electrophoresis patterns of pepsin-modified proteins showed mainly disappearances of minor protein bands, but no changes of main protein bands. The gel filtration patterns through Sephadex G-75 of sardine myofibrillar protein showed two big peaks and three small peaks. All the small peaks were disappearanced by proteolysis with pepsin in one hour. and during the period of proteolysis the fast big peak became gradually smaller and the late big peak eluted more slowly. By proteolysis, the emulsifying activity and emulsifying capacity of sardine myofibrillar protein were all decreased. The effects of pepsin-modification on emulsifying capacity were greater than those on emulsifying activity of protein. The aeration capacity of the protein was increased about 1.9 folds and the foam stability decreased to 0.6 folds of control by pepsin-modification. The pepsin-modified sardine myofibrillar proteins showed about 0.6 folds of heat coagulation and 1.4 folds of viscosity of control. The pH dependence of solubilities of sardine myofibrillar protein showed two isoelectric areas of pH 5 and 9. The pepsin-modified protein showed more clear pH dependences at the early stage but not at the late stage of proteolysis.

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Kinetic Studies of Lactic Acid Fermentation (Part 3) Effect of Phenol Derivatives on Fermentation (유산균발효에 관한 동력학적 연구 (제3보) 발효에 미치는 Phenol 유도체의 영향)

  • LEE Keun-Tai;YANG Hyeun-Suk
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.14 no.4
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    • pp.212-216
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    • 1981
  • The growth of Lactobacillus bulgaricus treated with vanillin, ortho-vanillin and guaiaco1 was studied on synthetic medium in mechanically agitated chemostat culture, The exponential-phase growth rate exhibited a maximum at the cells treated with 50 ppm vanillin. That stimulation, however, appears to be an effect on growth rate rather than total cell growth. And the others were inhibited by the chemicals. Much greater inhibition in growth of the cells treated with 100 ppm of each chemical than oars treated with 50 ppm was observed after 25 hour fomentation. For aerobic microbes, the alcohol dehydrogenase reaction is enhanced for the reproduction of NAD, which consequently cause to stimulate fermentation. For micro-aerophilic microbes , however, the same effect was not observed at the present study at least in the case of cell concentration. However except f or one treated with 50 ppm vanillin the same effect was observed in the case of growth is to. From the result using the glucose as a substrate, it was found that the cell concentrations measured in terms of ultimate optical density (UOB/ml), were 0.96 and 0.92, when treated with 50 and 100 ppm vanillin; 0.40 and 0.45 when treated with ortho-vanillin 50 and 100 ppm: 0.49 and 0.47, when treated with guaiacol 50 and 100 ppm. The specific growth rates were 0.44, 0.15, 0.25, 0.29, 0.37, and 0.34; the specific production rates wire 0.33, 0.15, 0.16, 0.22, 0.28, and 0.26 and the glucose concentrations (g/1) after 25 hour fermentation were 23.5, 32.8, 31.5, 29.5, 28.0 and 28.8, these all in the same sequences as the first.

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Influences of Saliva Substitutes on Salivary Enzymatic Activity (타액대체제가 타액 효소 활성에 미치는 영향)

  • Kho, Hong-Seop;Lee, Sung-Woo
    • Journal of Oral Medicine and Pain
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    • v.34 no.3
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    • pp.227-235
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    • 2009
  • Many of the protective functions of saliva can be attributed to the biological, physical, structural, and rheological characteristics of salivary glycoproteins. Therefore, the development of ideal saliva substitutes requires understanding of the rheological as well as biological properties of human saliva. In the present study, we investigated the changes of salivary enzymatic activities by saliva substitutes and compared viscosity of saliva substitutes with human saliva. Five kinds of saliva substitutes such as Moi-Stir, Stoppers4, MouthKote, Saliva Orthana, and SNU were used. Lysozyme activity was determined by the turbidimetric method. Peroxidase activity was determined with an NbsSCN assay. $\alpha$-Amylase activity was determined using a chromogenic substrate, 2-chloro-p-nitrophenol linked with maltotriose. The pH values of saliva substitutes were measured and their viscosity values were measured with a cone-and-plate digital viscometer at six different shear rates. Various types of saliva substitutes affected the activities of salivary enzymes in different ways. Stoppers4 enhanced the enzymatic activities of hen egg-white lysozyme, bovine lactoperoxidase (bLP), and $\alpha$-amylase. Saliva Orthana and SNU inhibited bLP activity and enhanced $\alpha$-amylase activity. MouthKote inhibited $\alpha$-amylase activity. Moi-Stir inhibited the enzymatic activities of bLP and $\alpha$-amylase. The pH values were very different according to the types of saliva substitutes. Stoppers4, MouthKote, and Saliva Orthana showed lower values of viscosity at low shear rates and higher values of viscosity at high shear rates compared with unstimulated and stimulated whole saliva. Moi-Stir and SNU displayed much higher values of viscosity than those of natural whole saliva. Collectively, our results indicate that each saliva substitute has its own biological and rheological characteristics. Each saliva substitute affects the enzymatic activity of salivary enzyme and finally oral health in different ways.

Metabolic Responses of Activated Sludge to Pentachlorophenol in a SBR System (SBR 처리 장치에서 활성 슬럿지의 대사에 미치는 Pentachlorophenol의 독성 효과)

  • KIM Sung-Jae;Benefield Larry D.
    • Journal of Aquaculture
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    • v.6 no.4
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    • pp.323-338
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    • 1993
  • The primary objective of this study was to examine the toxic effects of PCP on activated sludge and to analyze its metabolic responses while treating wastewater containing pentachlorophenol (PCP) in a sequencing batch reactor (SBR) system operating under different control strategies. This study was conducted in two phases 1 and 2 (8-hr and 12-hr cycles). Each phase was operated with two control strategies I and II. Strategy I (reactor 1) involved rapid addition (5 minutes to complete) of substrate to the reactor with continuous mixing but no aeration for 2 hours. Strategy II (reactor 2) involved adding the feed continuously during the first 2 hours of the cycle when the system was mixed but not aerated. During both phases each reactor was operated at a sludge age of 15 days. The synthetic wastewater was used as a feed. The COD of the feed solution was about 380 mg/L. After the reference response for both reactors was established, the steady state response of each system was established for PCP feed concentrations of 0.1 mg/L, 1.0 mg/L, and 5.0 mg/L in SBR systems operating on both 8-hr and 12-hr cycles. Soluble COD removal was not inhibited at any feed PCP concentrations used. At 5.0 mg/L feed PCP concentration and in SBR systems operating on phase 2, the concentrations or ML VSS were decreased; selective pressure on the mixed biomass might be increased, narrowing the range of possible ecological responses; the settleability of activated sludge was poor; the SOURs were increased, showing that the systems were shocked. Nitrification was made to some extent at all concentrations of feed PCP in SBR systems operating on phase 2 whereas in SBR systems operating on phase 1 little nitrification was observed. Then, nitrification will be delayed as much as soluble COD removal is retarded due to PCP inhibition effects. Enhanced biological phosphorus removal occurring in the system operating with control strategy I during phase 1 of this work and in the presence of low concentrations of PCP was unreliable and might cease at anytime, whereas enhanced biological phosphorus removal occurring in the system operating with either control strategy I or II during phase 2 of this work and in the presence of feed PCP concentrations up to 1.0 mg/L was reliable. When, however, such processes were exposed to 5.0 mg/L PCP dose, enhanced phosphorus removal ceased and never returned.

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Comparative Study on the Sawdust Cultivation and the Antioxidants of Hericium spp. (노루궁뎅이버섯류의 톱밥재배와 항산화물질 비교 연구)

  • Ryu, Sung-Ryul;Lee, Wi-Young;Ka, Kang-Hyeon
    • The Korean Journal of Mycology
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    • v.37 no.1
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    • pp.80-85
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    • 2009
  • As a preliminary study in order to develop new varieties of Hericium species, this study was carried out to investigate the optimal temperature for mycelial growth, to figure out the applicability to sawdust cultivation on Quercus mongolica substrate, and to analyze the antioxidant capacity of ergothioneine and polyphenols in Hericium strains preserved in Korea Forest Research Institute (KFRI). In the results of optimal temperature for mycelial growth of eight Hericium erinaceus, it was $20^{\circ}C$ in a strain (KFRI 842), $25^{\circ}C$ in five strains (KFRI 507, 508, 509, 843, 845), and $30^{\circ}C$ in two strains (KFRI 582, 844). Optimal temperature for mycelial growth of H. coralloides (KFRI 713) was $25^{\circ}C$. Four strains (KFRI 508, 843, 844, 713) out of the total nine Hericium strains showed full mycelium growth within 20 days at the optimal temperature on PDA medium in petri-dish (85 mm in diameter). The other strains have need of more time for full mycelium growth. Mushroom production of H. erinaceus ranged from 215 to 384 g of fresh weight and its dry weight was 7 to 9% of it, whereas that of H. coralloides was 299 g of fresh weight and its dry weight was 10% of it. The contents of ergothioneine and polyphenols of H. erinaceus strains were different by strains and those were in the range of $1.6{\sim}3.7$ mg/g dw. and $5.9{\sim}7.8$ mg/g dw., respectively. On the other hand, those of H. coraloides were in the range of 1.7 mg/g dw. and 3.9 mg/g dw., respectively. From the results of correlation ($R^2$ = 0.1) between ergothioneine and polyphenols in the strains, it was found that the total contents of them differ by strains but the ratio of the two compounds was not very different in the strains.

Optimum Synthesis Conditions of Coating Slurry for Metallic Structured De-NOx Catalyst by Coating Process on Ship Exhaust Gas (선박 배연탈질용 금속 구조체 기반 촉매 제조를 위한 코팅슬러리 최적화)

  • Jeong, Haeyoung;Kim, Taeyong;Im, Eunmi;Lim, Dong-Ha
    • Clean Technology
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    • v.24 no.2
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    • pp.127-134
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    • 2018
  • To reduce the environmental pollution by $NO_x$ from ship engine, International maritime organization (IMO) announced Tier III regulation, which is the emmision regulation of ship's exhaust gas in Emission control area (ECA). Selective catalytic reduction (SCR) process is the most commercial $De-NO_x$ system in order to meet the requirement of Tier III regulation. In generally, commercial ceramic honeycomb SCR catalyst has been installed in SCR reactor inside marine vessel engine. However, the ceramic honeycomb SCR catalyst has some serious issues such as low strength and easy destroution at high velocity of exhaust gas from the marine engine. For these reasons, we design to metallic structured catalyst in order to compensate the defects of the ceramic honeycomb catalyst for applying marine SCR system. Especially, metallic structured catalyst has many advantages such as robustness, compactness, lightness, and high thermal conductivity etc. In this study, in order to support catalyst on metal substrate, coating slurry is prepared by changing binder. we successfully fabricate the metallic structured catalyst with strong adhesion by coating, drying, and calcination process. And we carry out the SCR performance and durability such as sonication and dropping test for the prepared samples. The MFC01 shows above 95% of $NO_x$ conversion and much more robust and more stable compared to the commercial honeycomb catalyst. Based on the evaluation of characterization and performance test, we confirm that the proposed metallic structured catalyst in this study has high efficient and durability. Therefore, we suggest that the metallic structured catalyst may be a good alternative as a new type of SCR catalyst for marine SCR system.

Present status and prospect for development of mushrooms in Korea

  • Jang, Kab-Yeul;Oh, Youn-Lee;Oh, Minji;Im, Ji-Hoon;Lee, Seul-Ki;Kong, Won-Sik
    • 한국균학회소식:학술대회논문집
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    • 2018.05a
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    • pp.27-27
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    • 2018
  • The production scale of mushroom cultivation in Korea is approximately 600 billion won, which is 1.6% of the Korean gross agricultural output. Annually, ca. 190,000 tons of mushrooms are harvested in Korea. Although the numbers of mushroom farms and cultivators are constantly decreasing, the total mushroom yields are increasing due to the large-scale cultivation facilities and automation. The recent expansion of the well-being trend causes increase in mushroom consumption in Korea: annual per capita consumption of mushroom was 3.9kg ('13) that is a little higher than European's average. Thus the exports of mushrooms, mainly Flammulina velutipes and Pleurotus ostreatus, have been increased since the middle of 2000s. Recently, however, it is slightly reduced. However, Vietnam, Hong Kong, the United States, the Netherlands and continued to export, and the country has increased recently been exported to Australia, Canada, Southeast Asia and so on. Canned foods of Agaricus bisporus was the first exports of the Korean mushroom industry. This business has reached the peak of the sale in 1977-1978. As Korea initiated trade with China in 1980, the international prices of mushrooms were sharply fall that led to shrink the domestic markets. According to the high demand to develop new items to substitute for A. bisporus, oyster mushroom (Pleurotus ostreatus) was received the attention since it seems to suit the taste of Korean consumers. Although log cultivation technique was developed in the early 1970s for oyster mushroom, this method requires a great deal of labor. Thus we developed shelf cultivation technique which is easier to manage and allows the mass production. In this technique, the growing shelf is manly made from fermented rice straw, that is the unique P. ostreatus medium in the world, was used only in South Korea. After then, the use of cotton wastes as an additional material of medium, the productivity. Currently it is developing a standard cultivation techniques and environmental control system that can stably produce mushrooms throughout the year. The increase of oyster mushroom production may activate the domestic market and contribute to the industrial development. In addition, oyster mushroom production technology has a role in forming the basis of the development of bottle cultivation. Developed mushroom cultivation technology using bottles made possible the mass production. In particular, bottle cultivation method using a liquid spawn can be an opportunity to export the F.velutipes and P.eryngii. In addition, the white varieties of F.velutipes were second developed in the world after Japan. We also developed the new A.bisporus cultivar "Sae-ah" that is easy to grown in Korea. To lead the mushroom industry, we will continue to develop the cultivars with an international competitive power and to improve the cultivation techniques. Mushroom research in Korea nowadays focuses on analysis of mushroom genetics in combination with development of new mushroom varieties, mushroom physiology and cultivation. Further studied are environmental factors for cultivation, disease control, development and utilization of mushroom substrate resources, post-harvest management and improvement of marketable traits. Finally, the RDA manages the collection, classification, identification and preservation of mushroom resources. To keep up with the increasing application of biotechnology in agricultural research the genome project of various mushrooms and the draft of the genetic map has just been completed. A broad range of future studies based on this project is anticipated. The mushroom industry in Korea continually grows and its productivity rapidly increases through the development of new mushrooms cultivars and automated plastic bottle cultivation. Consumption of medicinal mushrooms like Ganoderma lucidum and Phellinus linteus is also increasing strongly. Recently, business of edible and medicinal mushrooms was suffering under over-production and problems in distribution. Fortunately, expansion of the mushroom export helped ease the negative effects for the mushroom industry.

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Parkin Interacts with the PDZ Domain of Multi-PDZ Domain Protein MUPP1 (Parkin과 Multi-PDZ Domain Protein (MUPP1) 단백질 간의 PDZ 결합)

  • Jang, Won Hee;Jeong, Young Joo;Choi, Sun Hee;Lee, Won Hee;Kim, Mooseong;Kim, Sang-Jin;Urm, Sang-Hwa;Moon, Il Soo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.24 no.8
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    • pp.820-826
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    • 2014
  • The localization to specific subcellular sites and the regulation of cell surface receptors and channels are crucial for proper functioning. Postsynaptic density-95/Disks large/Zonula occludens-1 (PDZ)-domain is involved in recognition of and interaction between various proteins, by which the localization and the regulation are mediated. Multi-PDZ domain protein 1 (MUPP1) contains 13 PDZ domains. MUPP1 serves a scaffolding function for structure proteins and signaling proteins, but the mechanism how MUPP1 is stabilized and signalized has not yet been elucidated. We used the yeast two-hybrid system to identify proteins that interact with PDZ domains of MUPP1. We found an interaction between MUPP1 and Parkin. Parkin is an E3 ubiquitin ligase. Loss-of-function mutations of Parkin gene are known to cause an autosomal recessive juvenile parkinsonism. Parkin bound to the $12^{th}$ PDZ domain, but not to other PDZ domains of MUPP1. The C-terminal end of Parkin has a type II PDZ-association motif, which was essential for the interaction with MUPP1 in the yeast two-hybrid assay. When co-expressed in HEK-293T cells, Parkin co-localized with MUPP1. When co-expressed with ubiquitin in HEK-293T cells, MUPP1 has been strongly ubiquitinated by Parkin. These findings collectively suggest that MUPP1 is a novel substrate of Parkin and its function or stability could be modulated by Parkin-mediated ubiquitination.

Metabolic Adjustments of Lactate Dehydrogenase Isozymes to the Environmental Temperature in Bluegill (Lepomis macrochirus) (환경온도에 대한 파랑볼우럭(Lepomis macrochirus) 젖산탈수소효소 동위효소들의 대사조절)

  • Ku, Bora;Cho, Sung Kyu;Yum, Jung Joo
    • Journal of Life Science
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    • v.26 no.10
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    • pp.1105-1112
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    • 2016
  • The aim of this study was to examine the metabolic adjustment of lactate dehydrogenase (EC 1.1.1.27, LDH) isozymes to the environmental temperature in bluegill (Lepomis macrochirus). This study included three groups of bluegill collected in April (group Ⅰ), May (group Ⅱ), and September (group Ⅲ). The LDH activities of skeletal muscle, heart, and brain tissues were higher in group Ⅲ than in groups Ⅰ and Ⅱ. The citrate synthase (EC 4.1.3.7, CS) activity was higher in skeletal muscle but lower in heart and brain tissues of group Ⅱ as compared to group Ⅰ. In contrast, the CS activity was lower in skeletal muscle and higher in heart and brain tissues in group Ⅲ than in group Ⅱ. Furthermore, the LDH/CS activity ratio was higher in the skeletal muscle and brain in group Ⅲ than in groups Ⅰ and Ⅱ. Accordingly, anaerobic metabolism was increased in group Ⅲ. LDH A4, A2B2, and B4 isozymes were expressed in skeletal muscle, heart, liver, and brain tissues. The LDH C hybrid was detected in brain tissue. The LDH A4 isozyme was successfully purified by affinity chromatography. The molecular weight of the purified LDH A4 isozyme was 136 kDa and its optimal pH for enzymatic activity was 8.0. The KmPYR values of LDH in skeletal muscle were 0.161-0.227 mM using pyruvate as a substrate. These kinetic properties of LDH in skeletal muscle are consistent with the fact that bluegill is a cold-adapted species. These results may be useful for predicting the habitat use of this fish.

The development of anti-DR4 single-chain Fv (ScFv) antibody fused to Escherichia coli alkaline phosphatase (대장균의 alkaline phosphatase가 융합된 anti-DR4 single-chain Fv (ScFv) 항체의 개발)

  • Han, Seung Hee;Kim, Jin-Kyoo
    • Korean Journal of Microbiology
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    • v.52 no.1
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    • pp.10-17
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    • 2016
  • Enzyme immunoassay to analyze specific binding activity of antibody to antigen uses horseradish peroxidase (HRP) or alkaline phosphatase (AP). Chemical methods are usually used for coupling of these enzymes to antibody, which is complicated and random cross-linking process. As results, it causes decreases or loss of functional activity of either antibody or enzyme. In addition, most enzyme assays use secondary antibody to detect antigen binding activity of primary antibody. Enzymes coupled to secondary antibody provide a binding signal by substrate-based color development, suggesting secondary antibody is required in enzyme immunoassay. Additional incubation time for binding of secondary antibody should also be necessary. More importantly, non-specific binding activity caused by secondary antibody should also be eliminated. In this study, we cloned AP isolated from Escherichia coli (E. coli) chromosome by PCR and fused to) hAY4 single-chain variable domain fragment (ScFv) specific to death receptor (DR4) which is a receptor for tumor necrosis factor ${\alpha}$ related apoptosis induced ligand (TRAIL). hAY4 ScFv-AP expressed in E. coli showed 73.8 kDa as a monomer in SDS-PAGE. However, this fusion protein shown in size-exclusion chromatography (SEC) exhibited 147.6 kDa as a dimer confirming that natural dimerization of AP by non-covalent association induced ScFv-AP dimerization. In several immunoassay such as ELISA, Western blot and immunocytochemistry, it showed antigen binding activity by color development of substrates catalyzed by AP directly fused to primary hAY4 ScFv without secondary antibody. In summary, hAY4 ScFv-AP fusion protein was successfully purified as a soluble dimeric form in E. coli and showed antigen binding activity in several immunoassays without addition of secondary antibody which sometimes causes time-consuming, expensive and non-specific false binding.