• Title/Summary/Keyword: Acid phosphatase-1

Search Result 511, Processing Time 0.028 seconds

Gene Cloning, Expression, and Characterization of Glucose-1-Phosphatase from Enterobacter cloacae B11

  • Kim, Young-Ok;Park, In-Suk;Nam, Bo-Hye;Kong, Hee-Jeong;Kim, Woo-Jin;Lee, Sang-Jun;Kim, Kyung-Kil
    • Fisheries and Aquatic Sciences
    • /
    • v.13 no.1
    • /
    • pp.49-55
    • /
    • 2010
  • A bacterial strain with phytase and glucose-1-phosphatase activity was isolated from seawater. The colony was identified as an Enterobacter cloacae strain and named E. cloacae B11. A gene, agpEnB11, coding for an intracellular acid glucose phosphatase was cloned from the strain and sequenced. It comprised 1,242 nucleotides and encoded a polypeptide of 413 amino acids. Recombinant glucose-1-phosphatase (AgpEn) was overexpressed in Escherichia coli and purified using Ni-NTA column under native conditions. Purified protein displayed a single band of 47 kDa on SDS-PAGE. AgpEn hydrolyzed a wide variety of phosphorylated compounds, with high activity for glucose-1-phosphate and glucose-6-phosphate. Optimum pH and temperature for enzyme activity were pH 5.0 and $50^{\circ}C$, respectively. Enzyme activity was stimulated by $Ca^{2+}$ and $Co^{2+}$, and inhibited by $Cu^{2+}$.

7,12-Dimethylbenz(a)anthracene에 의한 흰주 골모세포유사세포의 악성형질전환과 특성에 관한 연구

  • Lee, Jin
    • The Journal of the Korean dental association
    • /
    • v.37 no.7 s.362
    • /
    • pp.517-529
    • /
    • 1999
  • 본 연구는 태령 19일된 백서 태자 두 개관에서 분리한 골모세포유사세포에 화학발암물질인 7,12-Dimethylbenz(a)anthracene (DMBA: 0.5 ㎍/ml) 및 tumor promotor인 12-O-tetradecanoyl-phorbol-13-acetate (TPA; 1.0 ㎍/ml)를 단독 혹은 복합 처리하여 PTRCC-DMBA, RCC-DMBA 및 RCC-DMBA-TPA 세포주를 확립시키고, 각 세포의 세포형태, 세포성장곡선, alkaline phosphatase와 acid phosphatase 활성 및 in vitro tumorigenicity를 연구하였다. 또한 c-myc, c-랜, c-jun, p53 및 Rb 유전자의 발현변화와 항암단백질인 p53 및 pRb 단백질의 발현변화를 관찰하여 골모세포유사세포가 악성형질전환되는 분자기전의 일단을 연구하고자 시행하였다. 본 실험에 사용한 모든 세포군에서 높은 aikaline phosphatase 활성과 낮은 acid phosphatase/alkaline phosphatase ratio를 보여 골모세포의 특성을 나타내었다. RCC-DMBA와 RCC-DMBA-TPA 세포는 정상세포나 PTRCC-DMBA에 비해 빠른 성장속도를 보였으며, 또한 SOFT AGAR상에서 colony를 형성하여 anchorage-independent growth를 나타내었다. 화학발암 물질로 악성변형된 세포들은 정상세포나 PTRCC-DMBA 세포에 비해 c-myc 유전자의 과발현이 관찰되었다. 정상세포에서 p53 유전자의 발현은 1.9 kb의 message만이 발현되었다. 그러나 화학발암물질로 형질전환된 세포에서는 1.9 kb message외에도 1.6 kb의 message가 더 발현되었으며, message의 양도 현저히 증가되었다. p53 단백질의 발현은 RCC-DMBA-TPA 세포에서 정상세포에 비해 현저히 감소하였으나, RCC-DMBA 세포에서는 유사한 경향을 보였다. Rb 유전자의 발현은 RCC-DMBA-TPA 세포에서만 현저히 감소하였으나, Rb 단백질의 발현은 정상세포에 비해 형질전환된 세포들에서 모두 현저히 감소되었고, 특히 RCC-DMBA-TPA 세포에서는 거의 발현되지 않았다. 이상의 결과에서 백서 태자 두 개관에서 분리한 골모세포유사세포는 화학발암물질인 DMBA에 의해 악성형질전환이 유도되었으며, c-myc의 과발현 및 p53과 Rb 단백질의 발현감소가 정상 골모세포유사세포의 악성변형과정에 밀접히 연관되어 있음을 시사한다.

  • PDF

Determination of Branched-Chain α-Keto Acid Dehydrogenase Activity in Rat Tissues

  • Kim, Hyun-Sook;Johnson, Wayne A.
    • BMB Reports
    • /
    • v.28 no.1
    • /
    • pp.12-16
    • /
    • 1995
  • The branched-chain ${\alpha}$-keto acid dehydrogenase (BCKAD) complex is a rate limiting enzyme which catalyzes the oxidative decarboxylation of branched-chain ${\alpha}$-keto acids. Numerous studies have suggested that BCKAD is subject to covalent modification in vitro via phosphorylation and dephosphorylation, which are catalyzed by a specific kinase and phosphatase, respectively. The biggest difficulty in the assay of BCKAD activity is to arrest the interconversion between the active and inactive forms. BCKAD activity was determined from fresh rat heart and liver tissues using homogenizing and assay buffers containing inhibitors of phosphatase and kinase. The results suggest that a radiochemical assay using ${\alpha}$-keto[1-$^{14}C$]-isovalerate as a substrate for the enzyme can be applied as a reliable method to determine in vitro enzyme activity with arrested interconversion between the active and inactive forms of the BCKAD complex.

  • PDF

L-Ascorbic Acid-2-Phosphate Mg염의 합성 및 응용

  • 양창모
    • Journal of the Society of Cosmetic Scientists of Korea
    • /
    • v.13 no.1
    • /
    • pp.29-35
    • /
    • 1987
  • Purely synthesized L-ascorbic acid 2 phosphate Mg salt (1 AsA PMg) improved the weak point of ascorbic acid which is easily decomposed in water solution. This compound is hydrolyzed with phosphatase of skin to corresponding ascorbic acid giving Vitamine C activities. The buffer solution of potassium acetate 0.5% and citric acid 0.005% and the sodium sulfite respectively showed good stabilizing effect of the AsA PMg solution. Compared to the other ascorbic acid derivatives the good solubility of AsA PMg gives broad application to cosmetic field.

  • PDF

Direct Colorimetric Assay of Microcystin Using Protein Phosphatase

  • Oh, Hee-Mock;Lee, Seog-June;Kim, Jee-Hwan;Park, Chan-Sun;Yoon, Byung-Dae
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • v.5 no.6
    • /
    • pp.418-421
    • /
    • 2000
  • A new direct colorimetric assay of microcystin in water and algal samples is proposed consisting of two procedures as follows: 1) the elimination of phosphorus in the sample and concentration of microcystin using a C(sub)18 cartridge, 2) the detection of the released phosphorus by the ascorbic acid method and determination of protein phosphatase (PP) inhibition by microcystin. The optimum amounts of phosphorylase ${\alpha}$ and PP-1 in 50 ${\mu}$L concentrated sample were 50$\mu\textrm{g}$/50${\mu}$L buffer and 1.0unit/50${\mu}$L buffer, respectively, for the best assay. The pH for the maximum activity of PP-1 was 8. The minimum detectable concentration for this method was about 0.02$\mu\textrm{g}$/L, which is sufficient to meet the proposed guideline level of 1$\mu\textrm{g}$ microcystin/L in drinking water. Consequently, it would seem that the proposed direct colorimetric assay using PP is a rapid, easy, and convenient method for the detection of microcystin in water and algal samples.

  • PDF

Comparison of enzyme cytochemical activities between rosetted cells in peripheral blood of Korean native cattle (한우(韓牛) 말초혈액내(末梢血液內) rosette 형성세포(形成細胞)의 효소화학적(酵素化學的) 염색성(染色性) 비교(比較))

  • Kim, Nyeun-su;Song, Hee-jong;Kim, Sun-jae;Seo, Ye-won
    • Korean Journal of Veterinary Research
    • /
    • v.37 no.1
    • /
    • pp.129-135
    • /
    • 1997
  • Peripheral blood mononuclear cells(PBMNC) of Korean native cattle rosetted with Korean goat erythrocytes(KGRBC) and blood monocytes were evaluated for four cytochemical reactions such as acid phosphatase(ACP), alkaline phosphatase-anti-boby(ALP-Ab), ${\alpha}$-naphthyl butyrate esterase(${\alpha}$-NBE) and peroxidase. The results obtained were as follows; In rosetted cells, the positivities of ACP in E AET-DeX, EA and EAC were 70.3%, 22.4% and 25.2%, those of ${\alpha}$-NB were 27.4%, 44.2% and 79.8%, and those of ALP-Ab were 9.5%, 88.3% and 91.5%, respectively. Whereas, the positivity for Peroxidase in monocytes was 100%. In non-rosetted (remained) cells, the positivities of ACP in E AET+DeX. EA and EAC were 41.4%, 57.2% and 61.9%, those of ${\alpha}$-NB were 38.6%, 16.5% and 18.9% and those of ALP-Ab were 98.2%, 5.3% and 6.3%, in order.

  • PDF

A comparison of the hydrolase activities of excretory-secretory products and somatic extracts from fish parasitic nematodes, Anisakis simplex sensu stricto and Anisakis pegreffii larvae (어류 기생성 선충 Anisakis simplex sensu stricto와 Anisakis pegreffii 유충의 excretory-secretory products 및 somatic extracts의 가수분해효소 활성 비교)

  • Jeon, Chan-Hyeok;Wi, Seong;Kim, Jeong-Ho
    • Journal of fish pathology
    • /
    • v.27 no.1
    • /
    • pp.25-33
    • /
    • 2014
  • Hydrolase activities of excretory-secretory products (ESP) and somatic extracts (SE) from Anisakis simplex sensu stricto (s.s.) and Anisakis pegreffii larvae were investigated by using API ZYM kit. In esterase group, acid phosphatase showed high activity from both of A. simplex (s.s.) and A. pegreffii. Esterase (C4) showed activity only from SE and A. simplex (s.s.) showed higher activity than A. pegreffii. Alkaline phosphatase, acid phosphatase and naphthol-AS-BI-phosphohydrolase showed higher activity in 3rd stage larvae than in 4th stage larvae of both species. In aminopeptidase group, only leucine arylamidase showed remarkable activity in SE of both anisakid species, and A. simplex (s.s.) SE showed higher activity than A. pegreffii SE. In glycosidase group, N-acetyl-${\beta}$-glucosaminidase, ${\alpha}$-mannosidase, ${\alpha}$-fucosidase showed higher activity in A. simplex (s.s.) than A. pegreffii, and 4th larvae showed higher activity than 3rd larvae. These differences in hydrolase activity of anisakid nematodes larvae are thought to be due to different metabolism such as growth, moulting, digestion and feeding.

Emerging Roles of Human Prostatic Acid Phosphatase

  • Kong, Hoon Young;Byun, Jonghoe
    • Biomolecules & Therapeutics
    • /
    • v.21 no.1
    • /
    • pp.10-20
    • /
    • 2013
  • Prostate cancer is one of the most prevalent non-skin related cancers. It is the second leading cause of cancer deaths among males in most Western countries. If prostate cancer is diagnosed in its early stages, there is a higher probability that it will be completely cured. Prostatic acid phosphatase (PAP) is a non-specific phosphomonoesterase synthesized in prostate epithelial cells and its level proportionally increases with prostate cancer progression. PAP was the biochemical diagnostic mainstay for prostate cancer until the introduction of prostate-specific antigen (PSA) which improved the detection of early-stage prostate cancer and largely displaced PAP. Recently, however, there is a renewed interest in PAP because of its usefulness in prognosticating intermediate to high-risk prostate cancers and its success in the immunotherapy of prostate cancer. Although PAP is believed to be a key regulator of prostate cell growth, its exact role in normal prostate as well as detailed molecular mechanism of PAP regulation is still unclear. Here, many different aspects of PAP in prostate cancer are revisited and its emerging roles in other environment are discussed.

Morphological Characteristics of Seed Coat and Enzyme Activity of Aleurone Layers in Soybeans (콩 종피의 형태적 특성과 호분층 세포의 효소활성 연구)

  • 박정화;김용욱
    • KOREAN JOURNAL OF CROP SCIENCE
    • /
    • v.33 no.2
    • /
    • pp.112-121
    • /
    • 1988
  • Soybean seed coat consisted of three layers, and the aleurone layer was attached to the innermost part of seed coat. It showed the different morphological characteristics with single cell layer compared with many cell layers in barley aleurone layer. The structural difference in aleurone cell among varieties was not detected in this experiment. The hole of middle part of hilum consisted of net formed material in order to pass water and gas. In the experiment, it was not studied whether the varieties with hard seed consist of the same structure or not, but the detailed study on this would be necessary. The activity of acid phosphatase showed a tendency to increase in process of imbibition in distilled water. There was no significant difference in the enzyme activity among the varieties tested, but the enzyme activity of Danyoupkong was slightly higher than that of Hwanggeumkong. In germinability, Danyoupkong is higher than Hwangkeumkong, so it might be attributed to the higher activity. There was no difference in acid phosphatase activity between released from the aleurone cell and accumulated in the aleurone cell from 6 to 12 hours incubation of the medium in the absence of GA$_3$, but the difference was detected after 12 hours incubation. And enzyme activity was the highest in the 18 hours incubation. The increase in the release of acid phosphatase from the aleurone cell would be considered as passive diffusive effect due to the increase in turgo pressure of aleurone cell. The acid phosphatase released from aleurone layer increased greatly after 24 hours incubation of the medium in the presence of GA$_3$ and the accumulation within the aleurone cell decreased linearly after 18 hours incubation. The result indicates that GA$_3$ enhance the rate of enzyme release from aleurone layer, suggests that the aleurone cell wall be digested by the introduction of GA and the digested wall act as the channels for enzyme release.

  • PDF

Effect of Gibberellic acid on Isozyme Pattern of Rice Plant (Gibberellic acid가 수도의 Isozyme pattern에 미치는 영향)

  • Park, W.M.;Lee, Y.S.;Son, E.R.
    • KOREAN JOURNAL OF CROP SCIENCE
    • /
    • v.29 no.1
    • /
    • pp.39-45
    • /
    • 1984
  • The present researches were carried out to investigate the effects of gibberellic acid on the appearance of isozyme patterns of esterase, phosphatase, amylase and peroxidase, also to investigate if there were any differences of the isozyme patterns among root, shoot and seeds of rice plants. It was noticed that the isozyme patterns of the above tested enzymes were differ among the organs, root, shoot and seed. The GA treated plants showed difference of esterase patterns in root from Akibare and the difference in shoot and root from Yushin, phosphatase patterns in root from Akibare. However, the GA did not affect isozyme patterns of amylase or peroxidase. The seed should be the suitable organ to study isozyme patterns for genetics or variety characterization of rice plant.

  • PDF