• Title/Summary/Keyword: aminopeptidase A

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Enzymatic Degradation of Leucine Enkephalin and $[D-Ala^2]$-Leucine Enkephalinamide in Various Rabbit Mucosa Extracts (토끼의 수종 점막 추출액중 로이신엔케팔린 및 [D-알라$^2]$-로이신엔케팔린아미드의 효소적 분해 특성)

  • Chun, In-Koo;Park, In-Sook
    • YAKHAK HOEJI
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    • v.38 no.5
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    • pp.530-543
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    • 1994
  • To study the feasibility of transmucosal delivery of leucine enkephalin (Leu-Enk) and $[D-ala^2]$-leucine enkephalinamide (YAGFL), their degradation extents and pathways in various rabbit mucosa extracts were investigated by high performance liquid chromatography. The degradation of Leu-Enk and YAGFL was observed to follow the first-order kinetics. The degradation half-lives of Leu-Enk in the nasal, rectal and vaginal mucosal extracts were 1.62, 0.37 and 1.12 hrs and those of YAGFL were 30.55, 9.70 and 6.82 hrs, respectively, indicating Leu-Enk was degraded in a more extensive and rapid manner than YAGFL. But the mucosal and serosal extracts of the same mucosa showed the similar degradation rates for both pentapeptides. The degradation was most rapid in the neutral pH and increasing concentrations of substrates retarded the degradation rates. The maior hydrolytic fragments of Leu-Enk were Des-Tyr-Leu-Enk and tyrosine, indicating the enzymatic hydrolysis by aminopeptidases. However, the data also suggested endopeptidases such as dipeptidyl carboxypeptidase and dipeptidyl aminopeptidase could play some role in the degradation of Leu-Enk. On the other hand, the hydrolytic fragments of YAGFL in all the mucosa extracts were mainly Tyr-D-Ala-Gly and Phe-Leu-Amide, demonstrating the hydrolytic breakdown by endopeptidases. The degradation pathways were further explored by concomitantly determining the formation of smaller metabolites of primary hydrolytic fragments of Leu-Enk and YAGFL in the mucosa extracts.

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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.

Early Development of Digestive Organs, Intestinal Microvilli Digestive Enzymes, and Hepatic Antioxidant Enzymes after Hatching in Korean Native Chicks (한국 재래계에서 초기 성장에 따른 소화기관 발달, 소장 미세융모의 소화 효소 및 간조직의 항산화 효소 발현)

  • Geun-Hui Nam;Young-Bin Lee;Sea-Hwan Sohn;In-Surk Jang
    • Korean Journal of Poultry Science
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    • v.51 no.2
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    • pp.107-116
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    • 2024
  • The study was conducted to examine age-related development of digestive organs, intestinal microvilli hydrolase, and hepatic antioxidant enzyme in Korean native chicks (KNC) aged from 0-d to 28-d of post-hatching. Body weight did not significantly increase from 0-d to 3-d-old, but after that remarkably increased from 3-d to 28-d-old (P<0.05). The relative weight (g/100 g of BW) of the proventriculus, gizzard, and liver was significantly higher at 3- and 7-d-old chicks than that of the other ages. The relative weight of the intestine, mucosal tissues, and pancreas was markedly developed at the ages of 3-, 14-, and (or) 21-d-old chicks (P<0.05). In the small intestine, the specific activities of maltase and sucrose were significantly higher at 14-d-old compared with the other ages (P<0.05). Leucine aminopeptidase activity showed a constant level from 0- to 28-d-old without significance. The specific activity of alkaline phosphatase was significantly higher at 0-d-old compared with the other ages (P<0.05). In the liver, the specific activities of superoxide dismutase, glutathione peroxidase, and glutathione S-transferase were shown to be lowest at 0-d-old, but they continued to increase as the age increased. The lipid peroxidation was significantly high at the age of 21-d (P<0.05), after that its level decreased at 28-d old. In conclusion, the KNC rapidly developed digestive organs and intestinal microvilli hydrolase activity from 3- to 14-d-old after hatching. Hepatic antioxidant enzyme activity continued to increase as the age increased after hatching, resulting in 28-d-old chicks showing the highest antioxidant enzyme activity in the KNC.

Development of a Recombinant Streptomyces griseus with sprA and sprB Genes for Proteolytic Enzyme Production (Streptomyces griseus IFO13350 유래 sprA 및 sprB 유전자를 이용한 Pretense 생산균주 개발)

  • Hwang Ji-Hwan;Lee Chang-Kwon;Lee Kang-Mu;Jo Byoung-Kee;Park Hae-Ryong;Hwang Yong-Il
    • Korean Journal of Microbiology
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    • v.41 no.1
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    • pp.87-92
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    • 2005
  • Pronase, a protease produced for commercial purpose by Streptomyces griseus, was composed of serine protease, alkaline protease, aminopeptidase and carboxypeptidase complex, and it has been widely used as anti-inflammatory drugs for human therapy. In this study, we developed a new integration vector, pHJ101 derived from pSET152, containing strong promoter, ermE, to overexpress a certain protease gene. Specific PCR primers for cloning of sprA (a gene for S. griseus protease A) and sprB (a gene for S. griseus protease B) genes were designed from the basis of nucleotide sequence in databases and amplified by PCR. Plasmid pHJ201 and pHJ202 were constructed by inserting of amplified each gene in a vector pHJ101. S. griseus HA and S. griseus HB were respectively obtained by conjugal process of a parent strain, S. griseus IFO 13350 with the recombinant Escherichia coli harboring plasmid pHJ201 or pHJ202. When protease activity was measured in flask cultivation, produced protease levels of S. griseus HA and S. griseus HB increased about 5.3 times and 5 times, respectively, more than that of parent strain. And, the constructed integrating plasmid pHJ101 was applicable for overexpression of a certain gene in Streptomyces sp.

Degradation of human immunoglobulins and cytotoxicity on HeLa cells by live Trichomonas vaginalis (질편모충 단백질분해효소의 세포독성 및 인체면역글로불린 분해능)

  • 민득영;류재숙
    • Parasites, Hosts and Diseases
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    • v.35 no.1
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    • pp.39-46
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    • 1997
  • The present study was undertaken to determine whether live T. uaginnlis degrades human secretory IgA, serum If and IgG molecules. Human immunoglobulins were exposed to live trophozoites, parasite Iysate, and excretory-secretory product (ESP) of T ucginnlis. To determine the fragmentation of immunoglobulins, the reaction sample was subjected to SDS-PAGE and EITB, and peroxidase conjugated antihuman IgA and IgG were used as probes. Live trophozoites degraded secretory IgA, serum IgA and IgG, and degradation were pressed forward by the prolongation of the incubation time and by increasing the number of trichomonads respectively. Also the Iysates and ESP of trichomonads degraded IgA and IgG. The cysteine and serine proteinase inhibitors such as I-64, antipain, iodoacetic acid, iodoacetamide, TLCK reduced the ability of cleaving immunoglobulins. The proteinase activity and cytotoxicity of T. uaginnlis to HeLa cells were decreased when live T. vusinalis was treated with metallo-proteinase inhibitor as well as cysteine and serine proteinase inhibitors. These results suggest that proteinase secreted from live T ucginclis may play a part role in host pathogenesis by T. uosinnlis, and the cleaving ability of host immunoglobulins by the proteinase may contribute as a one of immune evasion mechanism for parasite survival in the host.

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Five Genes Regulated by Oryctes rhinoceros nudivirus Infection in the Intestinal Tube of Allomyrina dichotoma (장수풍뎅이(Allomyrina dichotoma)에 Oryctes rhinoceros nudivirus 감염에 의해서 유전자 발현이 조절되는 5개의 유전자)

  • Yoo, Bo-Kyung;Kwon, Kisang;Ko, Young Hwa;Lee, Eun Ryeong;Choi, Ji-Young;Kwon, O-Yu
    • Journal of Life Science
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    • v.26 no.11
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    • pp.1336-1340
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    • 2016
  • Korean rhinoceros beetles (Allomyrina dichotoma), which can be found in broad-leaved forests in mountainous habitats and lives for around one year in wild. This beetle is currently popular as a pet and traditionally regarded as a medicine for liver-related diseases in Korea. It is reported that the economic losses in the mass-rearing facilities by virus infection have been increased since the 2010s in Korea. The causing virus for the A. dichotoma was firstly reported as an Oryctes rhinoceros nudivirus (OrNV) in 2015. We, here, observes that serious morphological changes in the intestinal tube from the OrNV-infected beetles, and report five genes, which are regulated by OrNV infection in the intestine; Krueppel-like factor 15 (Klf15), Endoplasmic reticulum aminopeptidase 2 (ERAP2), U5 small nuclear ribonucleoprotein 200 kDa helicase (Snrnp200), Muscleblind-like protein 2a (mbnl2a), and MIMI_L93. The results may provide a clue to the early diagnosis and disease treatment during the mass-rearing facilities of the A. dichotoma.

Effects of a lipid-encapsulated zinc oxide supplement on growth performance and intestinal morphology and digestive enzyme activities in weanling pigs

  • Jang, Insurk;Kwon, Chang Hoon;Ha, Duck Min;Jung, Dae Yun;Kang, Sun Young;Park, Man Jong;Han, Jeong Hee;Park, Byung-Chul;Lee, Chul Young
    • Journal of Animal Science and Technology
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    • v.56 no.8
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    • pp.29.1-29.6
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    • 2014
  • This study compared the effects of varying lipid content and dietary concentration of a lipid-encapsulated (LE) ZnO product to those of native ZnO and thereby to find insights into optimal lipid coating and dosage of the Zn supplement. A total of 192 21-d-old weanling pigs were allotted to 48 pens, after which each six pens received a ZnO-free basal diet supplemented with 125 ppm ZnO (100 ppm Zn; BASAL), 2,500 ppm Zn as native ZnO (HIGH), or 100 or 200 ppm Zn as LE ZnO (LE-100 or LE-250) containing 8%, 10%, or 12% lipid [LE-8%, LE-10%, or LE-12%, respectively; $2{\times}3$ factorial arrangement within the LE-ZnO diets (LE-ALL)] for 14 d. Forty pigs were killed at the end for histological and biochemical examinations. None of ADG, ADFI, gain:feed, and fecal consistency score differed between the LE-ALL and either of the BASAL and HIGH groups. Hepatic and serum Zn concentrations were greater (p <0.05) in the HIGH vs. LE-ALL group, but did not differ between LE-ALL and BASAL, between LE-100 and -250, or among LE-8%, -10%, and -12% groups. Villus height (VH), crypt depth (CD), and the VH:CD ratio in the duodenum, jejunum, and ileum did not differ between the LE-ALL and either of the BASAL and HIGH groups, except for a greater CD in the duodenum in the LE-ALL vs. HIGH group. Additionally, VH and CD in the duodenum and VH:CD in the jejunum were greater in the LE-250 vs. LE-100 group. Specific activities of sucrase, maltase, and leucine aminopeptidase in these intestinal regions and those of amylase and trypsin in the pancreas were not influenced by the lipid content or dietary concentration of LE ZnO and also did not differ between the LE-ALL and either of the BASAL and HIGH groups, except for a greater pancreatic amylase activity in the former vs. HIGH group. In conclusion, the present results indicate that the LE ZnO, regardless of its lipid percentage or supplementation level examined in this study, has no significant effect on growth performance, fecal consistency, or digestive enzyme activities of weanling pigs under the experimental conditions.

Intestinal Growth and Development of Weanling Pigs in Response to Dietary Supplementation of Antibiotics, Phytogenic Products and Brewer's Yeast plus Bacillus Spores

  • Lee, C.-Young;Lim, Jung-Won;Ko, Young-Hyun;Kang, Sun-Young;Park, Man-Jong;Ko, Tae-Gu;Lee, Ji-Hoon;Hyun, Young;Jeong, Kyu-Sik;Jang, In-Surk
    • Journal of Animal Science and Technology
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    • v.53 no.3
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    • pp.227-235
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    • 2011
  • A total of 96 crossbred weanling barrows aged 21 days were randomly allocated to 32 pens of a new nursery to investigate the effects of antibiotics, phytogenics, and probiotics on intestinal growth and development. The animals were fed a set of three-phase basal diets containing 0.3% zinc oxide (CON) or the basal diets supplemented with 353 ppm of a combination of tiamulin, neomycin, chlortetracycline, and oxytetracycline (ANTI), 75 ppm triterpenoid saponin plus 150 ppm mixed saccharides (HERB; Sacchapin$^{(R)}$), or $1{\times}10^7$ brewer's yeasts plus $8{\times}10^7$ spores of each of Bacillus licheniformis and Bacillus subtilis per kilogram feed (PROBIO; Yeasture Plus 2B$^{(R)}$) for five weeks. Thirty-two pigs representing as many pens were slaughtered at the end of the feeding trial, after which morphological measures and digestive enzyme activities of intestinal mucosa were determined. Weight gain and gain:feed of the pigs were not affected by the dietary treatments (TRT) during the overall feeding trial. Total intestinal length was greater in PROBIO than in ANTI (P<0.05). Wet mucosa weight of the duodenum was not affected by TRT. However, jejunal mucosa weight was greater in PROBIO than in any other group sum of mucosa weights of the duodenum and jejunum was greater (P<0.05) in PROBIO than in ANTI and HERB. The height and width of duodenal villus were not affected by TRT, but crypt depth decreased (P<0.05) in response to HERB and PROBIO vs CON. Specific activities of alkaline phosphatase, sucrase, maltase, lactase, and leucine aminopeptidase in the duodenum and jejunum were not changed by TRT. In conclusion, results suggest that the present dietary treatments have no effects on growth performance of weanling pigs and that of PROBIO enhances intestinal growth and development under a clean experimental setting.

Observations on the Genetic Structure of Pinus densiflora Sieb. et Zucc(I) : The Young-il Population (소나무의 유전적(遺傳的) 구조(構造)에 관한 연구(硏究) (I) : 영일(迎日) 집단(集團)의 유전적(遺傳的) 구조(構造))

  • Chung, Min Sup
    • Journal of Korean Society of Forest Science
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    • v.80 no.2
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    • pp.246-254
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    • 1991
  • Genetic structure of a Pinus densiflora population consisting of two subpopulations on the north-and south-facing slopes of a mountain was studied by allozyme analysis. Allozyme variants in aspartate aminotransferase(AAT), glutmate dehydrogenase(GDH) and leucine aminopeptidase(LAP) systems are encoded, at least, by eight loci ; five for AAT, one for GDH and two for LAP. Average number of alleles examined over six loci was 3.33. Average heterozygosity and genetic diversity computed over six loci were, respectively, 0.19 and 2.76 for parental population, 0.17 and 2.22 for progeny population. Differences in allelic frequencies between maternal sources at many of the investigated loci were found and between subpopulations on the north- and south-facing slopes. Allele frequencies of maternal origin at some of the loci were significantly different from each other between the two subpopulations. Thus it appears that the matings within and between subpopulations were not random and the mountain ridge that divides the north-and south-facing slopes isolate the two suhpopulations reproductively to a great extent. Some of the genotypes both in parental and progeny(embryo) groups deviate significantly from the Hardy-Weinberg equilibrium state. It appears from the result that the pine population is originated from a few limited ancestral trees and thus consanguineous matings are prevalent in this pine population.

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Regulatory Mechanisms of Angiotensin II on the $Na^+/H^+$ Antiport System in Rabbit Renal Proximal Tubule Cells. I. Stimulatory Effects of ANG II on $Na^+$ Uptake

  • Han, Ho-Jae;Koh, Hyun-Ju;Park, Soo-Hyun
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.4
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    • pp.413-423
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    • 1997
  • The importance of the kidney in the development of hypertension was first demonstrated by Goldblatt and his colleagues more than fifty years ago. Many hormones and other regulatory factors have been proposed to play a major role in the development of hypertension. Among these factors angiotensia II (ANG II) is closely involved in renal hypertension development since it directly regulates $Na^+$ reabsorption in the renal proximal tubule. Thus the aim of the present study was to examine signaling pathways of low dose of ANC II on the $Na^+$ uptake of primary cultured rabbit renal proximal tubule cells (PTCs) in hormonally defined seum-free medium. The results were as follows: 1) $10^{-11}$ M ANG II has a significant stimulatory effect on growth as compared with control. Alkaline phosphatase exhibited significantly increased activity. However, leucine aminopeptidase and ${\gamma}-glutamyl$ transpeptidase activity were not significant as compared with control. In contrast to $10^{-11}$ M ANG II stimulated $Na^+$ uptake $(108.03{\pm}2.16% of that of control)$, $10^{-9}$ M ANG II inhibited ($92.42{\mu}2.23%$ of that of control). The stimulatory effect of ANG II on $Na^+$ uptake was amiloride-sensitive and inhibited by losartan (ANG II receptor subtype 1 antagonist) and not by PD123319 (ANG II receptor subtype 2 antagonist). 2) Pertussis toxin (PTX) alone inhibited $Na^+$ uptake by $85.52{\pm}3.52%$ of that of control. In addition, PTX pretreatment prevented the AMG II-induced stimulation of $Na^+$ uptake. 8-Bromoadenosine 3',5'-cyclic monophosphate (8-Br-cAMP), forskolin, and isobutylmethylxanthine (IBMX) alone inhibited $Na^+$ uptake by $88.79{\pm}2.56,\;80.63{\pm}4.38,\;and\;84.47{\pm}4.74%$ of that of control, respectively, and prevented the ANG II-induced stimulation of $Na^+$ uptake. However, $10^{-11}$ M ANG II did not stimulate cAMP production. 3) The addition of 12-O-te-tradecanoylphorbol-13-acetate (TPA, 0.01 ng/ml) to the PTCs produced significant increase in $Na^+$ uptake ($114.43{\pm}4.05%$ of that of control). When ANG II and TPA were added together to the PTCs, there was no additive effect on $Na^+$ uptake. Staurosporine alone had no effect on $Na^+$ uptake, but led to a complete inhibition of ANG II- or TPA-induced stimulation of Na'uptake. ANG II treatment resulted in a $111.83{\mu}4.51%$ increase in total protein kinase C (PKC) activity. In conclusion, the PTX-sensitive PKC pathway is the main signaling cascade involved in the stimulatory effects of ANG II on $Na^+$ uptake in the PTCs.

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