• Title/Summary/Keyword: Leucine aminopeptidase

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Activities of brush border membrane bound enzymes of the small intestine in iwetagonimus yokogatvai infection in mice (요꼬가와흡충에 감염된 마우스 소장의 미소융모막 효소 활성도)

  • 홍성태;유재란
    • Parasites, Hosts and Diseases
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    • v.29 no.1
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    • pp.9-20
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    • 1991
  • The present study intended to evaluate the influences of MetagonimMi yokogawai on the activities of brush border membrane bound enzymes of the small intestine. Mice were infected with 500 metacercariae respectively, and the worm recovery, morphological changes and enzyme activities were observed chronologically. A part of them were followed after the treatment. Recovered worms decreased in number continuously after the infection, and they were less than 10% after 2 weeks and almost Bero after 28 weeks. Villous atrophy and stromal inflammation were found at two locations of the proximal jejunum from 2 weeks to 4 weeks after the infection. The enzymes, alkaline phosphatase, leucine aminopeptidase and disaccharidases (sucrase, lactase, maltase, and trehalase), showed lowered activities in the duodenum and proximal jejunum of the infected mice but they increased in the distal jejunum for the first two weeks. From three weeks after the infection, the activities were gradually recovered. In one week treated mice, they recovered the activities at 2 weeks from the treatment, but there found no differences of the activities between the 3 week treated group and infected controls. The present data reveal that M. yckogawai infection induces degenerative changes of the host's intestinal mucosa not only morphologically but functionally during the initial phase of infection. The lowered enzyme activities in acute metagonimiasis should be associated with malabsorption and diarrhea.

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Isozyme Variability in Three Species of Freshwater Planorbid Snails in Korea : Gyraulus convexiusculus , Hippeutis cantori and Segmentina hemisphaerula (한국산 또아리물달팽이과 ( family Planorbidae ) 3종에서의 동위 효소 변이)

  • 정평림;정영헌;김기선
    • The Korean Journal of Malacology
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    • v.11 no.1
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    • pp.51-61
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    • 1995
  • A horizontal starch gal electrophoresis for enzyme proteins extracted from 3 species of Korean planorbid snails; Gyraulus convexiusculus, Hippeutis cantori and Segmentina hemisphaerula was carried out in order to elucidate their genetic relationships.The results from 12 enzymes employed in three different kinds of buffer systems are summarized as follows:1) Two loci from each enzyme of aldehyde oxidase, esterase, glucose phosphate isomerase. isocitrate dehydrogenase, leucine aminopeptidase, malate dehyogenase, peptidase and xanthine oxidase were detected, and only one locus was observed from each of the following four enzymes: 6-phosphogluconate dehydrogenase, glyceraldehyde-phosphate dehydrogenase, glutamate oxaloacetate transaminase and glycerol-3-phosphate dehydrogenase.2) Most of loci in 3 species of planorbid snails employed showed homozygous and monomorphic banding patterns and some of them were specifis as genetic markers among different species. However, a few of loci (EST-1. EST-2 and GPI-2)showed polymorphic banding patterns. 3)Hippeutis cantori and Segmentina hemisphaerula were more closely clustered in a dendrogram with the genetic iddentity value of 0.431, and these two species were lineated with Gyraulus convexiusculus as another cluster at the value of 0.294.In summarizing the above results, three species of Korean planorbid snails employed in this study mostly showed monomorphic enzyme protein banding patterns and genetic differences specific among 3 species.

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Effects of Insulin and IGFS on Growth and Functional Differentiation in Primary Cultured Rabbit Kidney Proximal Tubule Cells -Growth and membrane transport-

  • Han, Ho-Jae;Park, Kwon-Moo
    • The Korean Journal of Physiology
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    • v.29 no.2
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    • pp.191-202
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    • 1995
  • The purpose of this study was to compare effects of insulin and IGFs on growth, apical membrane enzyme activities and membrane transport systems of primary cultured rabbit kidney proximal tubule cells. Results were as follows: 1. Insulin and IGF-I produced significant growth stimulatory effects at $5{\times}10^{-10}M.\;IGF-II(5×10^{-10}\;M)$ did not stimulate significant cell growth. 2. Insulin stimulated the phosphorylation of a 97 KD protein. It was difficult to determine whether this band represents insulin and/or the IGF-I receptor. 3. The activities of apical membrane enzymes (alkaline phosphatase, leucine aminopeptidase, and ${\gamma}-glutamyl \;transpeptidase)$ were observed to be diminished after the cells were placed in the culture environment. 4. The uptake of ${\alpha}-MG,$ Pi and Na was significantly increased in cells incubated with insulin or IGF-I, IGF-II had no effect on the uptake of these substrates. 5. Na-pump activity, as assayed by Rb uptake, was significantly increased in cells treated with insulin or IGFs. In conclusion, insulin and IGF-I exert stimulatory effects on growth and membrane transporter(glucose, Na, Pi, and Na-pump) activities in primary cultured rabbit kidney proximal tubule cells. IGF-II had no effect on cell growth and membrane transporter(glucose, Na and Pi) activities.

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Studies on the Genetic Relationships of Sheep Populations from East and South of Central Asia

  • Sun, W.;Chang, H.;Yang, Z.P.;Geng, R.Q.;Lu, S.X.;Chang, G.B.;Xu, W.;Wang, H.Y.;Ren, Z.J.;Tsunoda, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.10
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    • pp.1398-1402
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    • 2002
  • Hu sheep was sampled randomly from Huzhou city, Zhejiang province, China. Of the 11 genetic markers from the blood examined by starch-gel and cellulose acetate electrophoresis, polymorphisms in Hu sheep were found for 10 loci, i.e. post-albumin (Po), transferring (Tf), alkaline phosphatase (Alp), leucine aminopeptidase (Lap), arylesterase (Ary-Es), hemoglobin-$\beta$ (Hb-$\beta$)、Xprotein(X-p), carbonic anhydrase (CA), catalase (Cat) and lysine (Ly). The same data except for Po locus were collected from another 14 sheep breeds from China and other countries, in order to ascertain their genetic relationships with one another and with the Hu sheep. The sheep populations from the east and south of Central Asia can be classified into three genetic groups: 'Mongolian sheep', 'South Asian sheep' and 'European sheep'. The Hu sheep belong to the 'Mongolian sheep' group.

The Purification and Characterization of Bacillus subtilis Tripeptidase (PepT)

  • Park, Yong-Seek;Cha, Myung-Hoon;Yong, Whan-Mi;Kim, Hyo-Joon;Chung, Il-Yup;Lee, Young-Seek
    • BMB Reports
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    • v.32 no.3
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    • pp.239-246
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    • 1999
  • A tripeptidase (PepT) was purified to homogeneity from Bacillus subtilis through four sequential chromatographies including DEAE-Sepharose ion exchange, hydroxylapatite, mono-Q FPLC ion exchange, and Superose-12 FPLC gel filtration. The apparent molecular mass of the enzyme was 49,200 Da and 51,400 Da as determined by sodium dodecylsulfatepolyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration chromatography, respectively, and the enzyme exists in a monomeric form. The physicochemical properties of the enzyme were as follows: optimum pH at 7.5, optimum temperature at $60^{\circ}C$, and pI at 4.9. The $K_m$ and $V_{max}$ values of the enzyme were 4.3 mM and 2.5 mmol/min/mg, respectively, with MetAla-Ser as substrate. The B. subtilis PepT requires $Co^{2+}$ ion(s) for activation, while it is inactivated by EOTA and 1,10-phenanthroline, suggesting that it is a metalloprotein. The enzyme was not inhibited by any of serine protease, aspartic protease, or leucine aminopeptidase inhibitors. The enzyme showed comparable activities towards four different substrates including Met-Ala-Ser, Leu-Gly-Gly, Leu-Ser-Phe, and Leu-Leu-Tyr. The amino terminal sequence of PepT determined by Edman degradation was found to be MKEEIIERFTTYVXV and turned out to be identical to that of PepT deduced from a cloned B. subtilis pepT.

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Expression of Antihypertensive Peptide, His-His-Leu, as Tandem Repeats in Escherichia coli

  • Jeong, Do-Won;Shin, Dong-Seok;Ahn, Chang-Won;Song, In-Sang;Lee, Hyong-Joo
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.952-959
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    • 2007
  • His-His-Leu (HHL), a tripeptide derived from a Korean soybean paste, is an angiotensin-I-converting enzyme (ACE) inhibitor. We report here a method of producing this tripeptide efficiently by expressing tandem multimers of the codons encoding the peptide in E. coli and purifying the HHL after hydrolysis of the peptide multimers. The HHL gene, tandemly multimerized to a 40-mer, was ligated with ubiquitin as a fusion gene (UH40). UH40 was inserted into vector pET29b; the UH40 fusion protein was then produced in E. coli BL21. The recombinant UH40 protein was purified by cation-exchange chromatography with a yield of 17.3mg/l and analyzed by matrixassisted laser desorption ionization (MALDI) time-of-flight (TOF) mass spectrometry and protein N-terminal sequencing. Leucine aminopeptidase was used to cleave a 405-Da HHL monomer from the UH40 fusion protein and the peptide was purified using reverse-phase high-performance liquid chromatography (HPLC) on a C18 HPLC column, with a final yield of 6.2mg/l. The resulting peptide was confirmed to be HHL with the aid of MALDI-TOF mass spectrometry, glutamine-TOF mass spectrometry, N-terminal sequencing, and measurement of ACE inhibiting activity. These results suggest that our production method is useful for obtaining a large quantity of recombinant HHL for functional antihypertensive peptide studies.

Selection of Highly Bitter taste Strains on Ganoderma sp. (영지버섯 고미성 균주 선발에 관한 연구)

  • Kim, Kyung-Soo;Byun, Myung-Ok;You, Chang-Hyun;Cha, Dong-Yeul;Jeong, Hoon;Ko, Mi-Suk
    • The Korean Journal of Mycology
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    • v.22 no.4
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    • pp.350-354
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    • 1994
  • A study was conducted on the characteristics of 11 strains of Ganoderma sp. to select the strains with more bitterness. Among the flat type strains, ASI 7071 and 7091 showed higher bitterness, while among the branched type strains ASI 7074 and 7094 were found to more bitter than other strains. The growth of ASI 7091 was best on Ganoderma complete media(GCM), while ASI 7010, 7048 and 7075 performed best in oak saw dust media. Among the branched type strains the esterase isozyme band patterns were similar. On the other hand among the flat type strains, the esterase isozyme band patterns differed from each other. Out of 11 strains, ASI 7004 was found to have the heaviest fruiting body, ASI 7071 the thickest cap and ASI 7094 the biggest cap.

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Enzymatic Activity and Amino Acids Production of Predominant Fungi from Traditional Meju during Soybean Fermentation

  • Dong Hyun Kim;Byung Hee Chun;Jae-Jung Lee;Oh Cheol Kim;Jiye Hyun;Dong Min Han;Che Ok Jeon;Sang Hun Lee;Sang-Han Lee;Yong-Ho Choi;Seung-Beom Hong
    • Journal of Microbiology and Biotechnology
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    • v.34 no.3
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    • pp.654-662
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    • 2024
  • To investigate the effect of the predominant fungal species from Korean traditional meju and doenjang on soybean fermentation, the enzymatic activity and amino acid production of twenty-two fungal strains were assessed through solid- and liquid-state soybean fermentation. Enzymatic activity analyses of solid-state fermented soybeans revealed different enzyme activities involving protease, leucine aminopeptidase (LAP), carboxypeptidase (CaP), glutaminase, γ-glutamyl transferase (GGT), and amylase, depending on the fungal species. These enzymatic activities significantly affected the amino acid profile throughout liquid-state fermentation. Strains belonging to Mucoromycota, including Lichtheimia, Mucor, Rhizomucor, and Rhizopus, produced smaller amounts of total amino acids and umami-producing amino acids, such as glutamic acid and aspartic acid, than strains belonging to Aspergillus subgenus circumdati. The genera Penicillium and Scopulariopsis produced large amounts of total amino acids and glutamic acid, suggesting that these genera play an essential role in producing umami and kokumi tastes in fermented soybean products. Strains belonging to Aspergillus subgenus circumdati, including A. oryzae, showed the highest amino acid content, including glutamic acid, suggesting the potential benefits of A. oryzae as a starter for soybean fermentation. This study showed the potential of traditional meju strains as starters for soybean fermentation. However, further analysis of processes such as the production of G-peptide for kokumi taste and volatile compounds for flavor and safety is needed.

Effects of Dietary Supplementation of Coffee Meal on Intestinal Enzyme Activity, Biochemical Profiles and Microbial Population in Broiler Chicks (커피박 첨가가 육계의 소장 효소 활성도, 생화학 지표 및 장내 미생물 균총에 미치는 영향)

  • Ko, Young-Hyun;Yun, Seo-Hyun;Song, Min-Hae;Kim, Se-Yun;Kim, Jong-Sun;Kim, Hyoun-Wook;Jang, In-Surk
    • Korean Journal of Poultry Science
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    • v.41 no.2
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    • pp.105-113
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    • 2014
  • The current study was performed to investigate the effects of dietary supplementation of dried coffee meal (CM) on growth performance, intestinal and blood biochemical index, intestinal enzymes, and cecal microbial populations. A total of 162, 3-day-old male broiler chicks were randomly allocated into three dietary groups: control group (CON), basal diet added with 0.5% CM (CM I), and basal diet added with 1.0% CM (CM II). Dietary supplementation of CM did not change bird performance and the relative weight of intestinal mucosal tissues. The birds fed the diet supplemented with CM (0.5 and 1.0%) significantly decreased mucosal glucose concentration (P<0.05) without affecting blood glucose level compared with those fed control diet. The level of blood aspartate aminotransferase (AST) significantly increased in CM II group (P<0.05) without affecting ${\gamma}$-glutamyl transpeptidase (${\gamma}$-GTP) compared with that in the CON group. The specific activity of intestinal maltase, leucine aminopeptidase (LAP) and alkaline phosphatase (ALP) were not affected by dietary supplementation of CM, whereas sucrase activity in birds fed the diet supplemented with CM was decreased (P<0.05) compared to that in the control birds. The colony forming units (CFU) of E. coli in the cecum of CM-fed birds was significantly decreased (P<0.05) compared with that of control birds without changing the CFU of Lactobacillus. In conclusion, dietary supplementation of lower level of CM (0.5%) can be used as a beneficial feed resource without liver toxicity in broiler chicks.

Effects of Probiotic Complex on Performance, Blood Biochemical and Immune Parameters, Digestive Enzyme Activity, Fecal Microbial Population and Noxious Gas Emission in Broiler Chicks (복합생균제가 육계의 생산성, 혈액생화학성분과 면역지표, 소화효소 활성도, 분중 미생물 및 유해가스 발생에 미치는 영향)

  • Kim, Min-Jeong;Jeon, Dong-Gyung;Ahn, Ho-Sung;Yoon, Il-Gyu;Moon, Eun-Seo;Lee, Chai-Hyun;Lim, Yong;Jang, In-Surk
    • Korean Journal of Poultry Science
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    • v.47 no.3
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    • pp.169-180
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    • 2020
  • This study examined the effects of a probiotic complex (PC) containing Lactobacillus plantarum, Bacillus subtilis, and Saccharomyces cerevisiae on growth performance, organ weight, immune parameters, fecal microbial count, and noxious odor in broiler chicks. A total of 216 birds (4-day-old) were fed a basal diet (CON) and a diet supplemented with 0.25% (PC1) and 0.5% (PC2) of PC until 35 days of age. No difference in body weight, feed intake, and FCR was observed among the groups. The intestinal mucosal weight of the PC1 group was greater than that of the CON group without affecting weights of the other organs. Intestinal secretory immunoglobulin A (sIgA) levels in the PC2 group increased significantly (P<0.05) compared with that in the CON group. The PC2 group also had a strong tendency for elevated blood sIgA levels. Dietary PC did not affect the level of interleukin-1β in the blood and mucosal tissues or alter maltase, sucrase, and leucine aminopeptidase activities in the intestinal mucosa. The PC2 group had higher colony-forming units (cfu) for L. plantarum and S. cerevisiae, but lower cfu for E. coli than those in the CON group. Compared to the CON diet, the PC2 diet resulted in a decreased H2S concentration and a tendency toward decreased CH3SH concentration. In conclusion, a 0.5% PC diet showed increased sIgA and desirable microbial population, and decreased noxious odor in the feces, suggesting that PC could be applied as an environmentally friendly feed additive in broiler chicks.