• Title/Summary/Keyword: Aminopeptidase

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The Structural Characterization of Recombinant Bovine Somatotropin Expressed in Escherichia coli (재조합 소성장호르몬의 구조적 특성)

  • 김정호;김훈주박은숙김준
    • KSBB Journal
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    • v.9 no.2
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    • pp.165-173
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    • 1994
  • In this paper we have described the structural characterization of recombinant bovine somatotropin produced in Escherichia coli. Recombinant bovine somatotropin consists of 191 amino acid residues with a calculated molecular weight of 21,802 Da. For fragmentation of recombinant bovine somatotropin, we have used trypsin, Staphylococcus aureus V8 pretease, CNBr, and mild acid hydrolysis method. Digestion and cleavage with these proteases and chemicals yielded peptides of various size for amino acid sequence determination. The N-terminal sequence analysis was carried out up to thirty residues. Because the design of the recombinant bovine somatotropin gene for expression was such that the coding sequence begins with an initiation codon, AUG, before Ala, the first amino acid of bovine somatotropin, we could expect the initial amino acid as N-formyl Met. But the first amino acid of this protein, expressed in E. coli cells as inclusion bodies, was Ala. And the amino acid composition of RP-HPLC purified recombinant bovine somatotropin was determined and no essencial difference was observed. The amino acid sequence of the recombinant bovine somatotropin was identical to that predicted from its recombinant gene. There was no processing or replacement of amino acid residues in recombinant bovine somatotropin expressed in E. coli. The hydropathy plot of recombinant bovine somatotropin revealed a hydrophobic region at the NH2-terminus and hydrophilic region at the COOH-terminus. The E. coli expression system is thought to be valuable for the expression of recombinant bovine somatotropin because protein was processed to remove the N-terminal Met residue by methionyl-aminopeptidase autonomously.

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Purification and Characterization of Authentic Human Growth Hormone Converted from Methionyl Human Growth Hormone by Immobilized Aminopeptidase M (고정화 Aminopeptidase M에 의해 메치오닐 인간성장호르몬으로부터 전환된 천연형 인간성장호르몬의 정제 및 특성 확인)

  • 이성희;조영우
    • KSBB Journal
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    • v.10 no.3
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    • pp.271-282
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    • 1995
  • The authentic hGH converted from met-hGH by immobilized ApM was purified by successive chromatographic processes based on the differences in isoelectric points, hydrophobicities and charges. The final recovery yield was about 14.1% and the specific activity of the purified hGH was 2.75IU per mg when assayed by enzyme immunoassay. The purified hGH was verified to be authentic hGH through the analysis of amino acid composition, amino-terminal amino acid sequence, carboxy-terminal amino acid and tryptic peptide map. The purity of purified hGH was higher than that of commercial hGH when assessed by SDS-PAGE, PAGE, IEF and HSGF. In weight-gain assay and tibia test with hypophysectomized rats, the hGH produced in this study showed the same growth effect as the commercial hGH.

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Effects of the Administration of 5-(4'- Pipweisinomwrhylphwnly)-2,3-dihydroimidazo[2,1-a] is pquinoline (SDZ-62-434) on Rat Kidney

  • Yi, E.Y.;Ma, Y.;Choi, W.J.;Park, J.S.;Cheon, S.H.;Lim, D.K.
    • Toxicological Research
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    • v.12 no.2
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    • pp.277-281
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    • 1996
  • To evaluate the renal toxicity of the antitumor agent, 5-(piperidonomethylphenyl)-2,3-dihydroimidazo[2,1-a]isoquinoline (SDZ-62-434), rats were treated with SDZ-62-434 of 50 mg/Kg, i.p., once and 10 mg/Kg, i.p., daily for 7 days. The kidney weights and urine volume after and during the treatment were observed. The concentrations of urinary creatinine, protein, and the activities of N-acetyl-$\beta $D-glucosaminidase (NAG), alanine aminopeptidase (AAP), $\gamma$-glutamyl transpeptidase (GGT) and lactate dehydrogenase (LDH) in 24 hr urine were also determined. The kidney weights after acute and subacute administration was not affected. The urine excretions were increased 5 days after the acute administration and increased after the daily 3rd day-administration. The excretion of creatinine was similar as that of urine excretion. The excretion of creatinine was increased 5 days after the acute and subacute administration. However, the protein excretion didn't changed in both treatment. Those indicate that SDZ-62-434 might induce the diuresis and also suggest that diuresis might be due to the some metabolites rather than the compound itself. The urinary activities of NAG and LDH were not affected after the acute treatment. However, the urinary activities of AAP and GGT were slightly increased 3 days after the acute administration but, returned to the control value. In subacute treatment, the activities of GGT was not changed. And the activities of NAG were declined after the 7th day-administration. However, the activities of AAP were significantly increased after the 5th day-administration. Furthermore, the urinary activities of LDH were continuously increased during the subacute administration. These results indicate that the high and subacute administration might induce a weak damage on the kidney cells. Furtherrnore, the present results suggest that SDZ-62-434 might have relatively slow-emerging and mild toxicity to the kidney.

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Genetic Analysis of Some Polymorphic Isozymes in Pinus densiflora (I) -Inheritance of Glutamate-Oxalate Transaminase and Leucine Aminopeptidase, and Linkage Relationship among Allozyme Loci- (소나무의 몇가지 다형적(多形的) 동위효소(同位酵素)의 유전분석(遺傳分析)(I) -Glutamate-Oxalate Transaminase 와 Leucine Aminopeptidase의 유전(遺傳)과 동위효소(同位酵素) 유전자좌(遺傳子座) 간(間)의 연관관계(連關關係)-)

  • Kim, Zin Suh;Hong, Yong Pyo
    • Journal of Korean Society of Forest Science
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    • v.58 no.1
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    • pp.1-7
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    • 1982
  • Megagametophyte and embryo tissue of Pinus densiflora were subjected to study the inheritance of glutamate-oxalate transaminase(GOT) and leucine aminopeptidase(LAP), and linkage relationship among isozyme loci coding both enzymes by starch gel zone-electrophoresis. Four zones of activity were observed for GOT. No variation was found in the fastest migrating zone (GOT-A). Electrophoretic phenotypes of the other two zones (GOT-B and GOT-C) showed 1:1 segregation ration, suggesting that each zone is controlled by a single locus. Foru and three alleles were identified at both loci respectively. The isozyme pattern of the fourth zone(GOT-D), migrated cathodally, coincided precisely with that of GOT-C. Whether the two zones are controlled by the same locus or by two tightly linked loci remained unknown. In all three variant GOT zones, heterozygoes embryos produced triple band patterns, indicating that GOT isozyme in Pinus densiflora is a dimer. Two zones of activity stained for LAP were found. The segregation of the two zones (LAP-A and LAP-B) suggested that tow loci control each of both isozymes. Two and three alleles were identified at both loci. GOT-B and LAP-B were found to be tightly linked, showing an average recombination frequency of 12.5 percent. Slight deviation from independent assortment was observed between GOT-B and GOT-C, with recombination frequency of 41 percent.

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Genetic Variation of the Pleurotus ostreatus Complex Based on Isozyme Analysis (동위효소 분석에 의한 Pleurotus ostreatus Complex의 유전적 변이)

  • Lee, Hee-Kyung;Yoo, Young-Bok;Min, Kyung-Hee
    • The Korean Journal of Mycology
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    • v.27 no.5 s.92
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    • pp.328-336
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    • 1999
  • Isozyme comparisons of mycelial extracts from Pleurotus ostreatus were undertaken using isoelectric focusing. Enzyme isozyme patterns were Used to describe the extent of geographical diversity and degree of intraspecific variation in these extracts. A total of 77 bands were resolved from six different enzymes. Cluster analyses were performed using the zymograms for esterase (EST), glucose phosphate isomerase (GPI), leucine aminopeptidase (LAP), malate dehydrogenase(MDH), peroxidase (POX), and phosphoglucomutase (pGM). EST gave multiple banding patterns, while less variability was observed for GPI, MDH, and PGM. Cluster analyses demonstrated that strains of P. ostreatus from geographically different origins are genetically divergent, supporting the idea that there is little or no gene flow between these geographically distant population groups.

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Preliminary Studies on the Relationship between Reed and Bacterial Communities in the Salt Marsh Environment of Namyang Bay, Korea

  • Kwon, Kae-Kyoung;Je, Jong-Geel
    • Ocean and Polar Research
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    • v.24 no.1
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    • pp.47-53
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    • 2002
  • To evaluate the effect of reed population on the distribution and activities of microorganisms, vertical distribution of heterotrophic bacteria, degradation rate of cellulose, extracellular aminopeptidase activity (APA) and metabolic diversity based on GN2 Microlog plate were measured at two salt marsh stations in Hogok-ri, Namyang Bay, west coast of Korea. The number of heterotrophic bacteria at station 1 (reed population inhabited area) showed 2 to 6 times higher than that of station 2 (exposed area) with exception in the surface layer. Cellulose degradation rates in station 1 showed more than 50%. month-I and higher than that of station 2 (10.2 to 38.4%. $month^{-1}$). Yet the APA at two stations did not show difference except surface layer and suggested that APA might not be a significant factor in degrading marsh plant debris. Lipid class compounds, cell wall polymers and L-alanine were widely used by microorganisms. The number and activities of bacterial populations especially concerned in plant debris degradation seemed to be stimulated by the reed communities.

Bioactive Metabolites from Selected Sponges of Korean and Tropical Waters

  • Shin, Jong-Heon;Park, Jung-Rae;Seo, Young-Wan;Lee, Hyi-Seung;Cho, Ki-Woong
    • Proceedings of the PSK Conference
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    • 2001.04a
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    • pp.90-94
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    • 2001
  • Wondonins A and B, aromatic alkaloids of an unprecedented skeletal class have been isolated form and association of the sponges Poecillastra wondoensis and Jaspis sp. In addition, four novel bromotyrosine-derived metabolites, psammaplins $A_1$ and $A_2$, aplysinellins A and B, have been isolated from the tropical sponge Aplysinella rhax. The structures of these compounds have been determined on the basis of combined chemical and spectral analyses. The new compounds exhibited significant cytotoxicity and antiangiogenic activity as well as inhibitory activities against farnesyl protein transferase and leucine aminopeptidase. In addition to these compounds, several bioactive metabolites have been isolated from sponges of Korean and tropical waters.

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Beta-amyloid peptide degradation by aminopeptidase and its functional role in Alzheimer's disease pathogenesis

  • AhnJo, Sang-Mee
    • 한국약용작물학회:학술대회논문집
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    • 2006.04a
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    • pp.75-90
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    • 2006
  • Both synthetic and endogenous $A{\beta}$ are degraded by peptidase G. Both $A{\beta}40$ and 42 are cleaved by peptidase G. Peptidase G cleaves $A{\beta}40$ into small fragments ($A{\beta}18$) which lacks aggregation property and are not toxic to neuron. Peptidase G seems to degrade multimeric $A{\beta}$ more efficiently than monomeric $A{\beta}$. Peptidase G protects neurons from toxicity induced by $A{\beta}$ by cleaving it into smaller fragments. Thus, dis-regulation of peptidase G could contribute amyloid deposit found in AD brain.

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Enzymatic Activities in Petroleum Wastewater Purification System by an Activated Sludge Process

  • Li Yin;Chrost Ryszard J.
    • Journal of Microbiology and Biotechnology
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    • v.16 no.2
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    • pp.200-204
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    • 2006
  • The enzymology of an activated sludge system for a petroleum wastewater purification process was investigated. Leucine-aminopeptidase (L-AMP), ${\beta}$-glucosidase (${\beta}-GLC$), and lipase (LIP) were selected for the study. It was found that more than 81.7% of enzymatic activity was associated with microbial cells in the activated sludge floc. The metabolic response of a mixed microbial population to increased phenol concentration showed that L-AMP activity increased in the activated sludge, whereas activities of ${\beta}-GLC$ and LIP decreased, due to the inhibitory effect of the phenol which varied from 100 mg/l to 500 mg/l.