• Asian-Australasian Journal of Animal Sciences
• /
• v.2 no.2
• /
• pp.99-102
• /
• 1989

The level of serum alkaline phosphatase activity was determined in 7 Japanese Black steers at different ages. The isoenzyme activity of non-bone origin was estimated using a heat-inactivation technique. The activity of serum alkaline phosphatase (SALP, K-A unit) decreased as age (AGE, mo.) increased: SALP = 14.15 - 0.17 (${\pm}\;0.03$) AGE, r = -0.81, P<0.01, $S.E.\;{\pm}\;0.28$. The variation of the activity was greater in younger age than the older. The temperature of $58^{\circ}C$ for the treatment of heat inactivation of bovine serum appeared to be suitable. The percentage of heat inactivated enzyme activity negatively correlated with age and positively with the level of serum alkaline phosphatase activity. The activity of SALP of non-bone origin was inferred to stay at about constant level irrespective of age and that of bone origin decreased with age.

• Journal of Microbiology and Biotechnology
• /
• v.13 no.5
• /
• pp.660-665
• /
• 2003

High-level expression of Thermus caldophilus GK24 alkaline phosphatase (Tca APase) was achieved in Escherichia coli using the pET-based expression plasmids, pEAP1 and pEAP2. In the case of plasmid pEAP2, the signal peptide region of Tca APase was replaced by the PelB leader peptide of expression vector pET-22b(+). Furthermore, the expression level was somewhat higher than that of plasmid pEAPl. A rapid purification procedure of Tca APase overproduced in E. coli was developed which involved heating to denature E. coli proteins followed by HiTrap Heparin HP column chromatography. Optimal temperature and pH and $Mg^{2+}$ dependence of the recombinant Tca APase were similar to those of native enzyme isolated from T. caldophilus GK24.

• BMB Reports
• /
• v.30 no.4
• /
• pp.262-268
• /
• 1997

The thermophilic and thermostable alkaline phosphatase was purified to near homogeneity from the osmotic lysis of Thermus caldophilus GK24, The purified enzyme had an apparent molecular mass of 108, 000 Da and consisted of two subunits of 54,000 Da. lsoelectric-focusing analysis of the purified enzyme showed a pi of 7.3. The enzyme contained two Cys residues, and its amino acids composition was quite different from that of Thermus aquaticus YT-1 alkaline phosphatase and Escherichia coli alkaline phosphatase, The optimum pH and temperature of the enzyme were 11.0-11.5 and $80^{\circ}C$ respectively. The enzyme was stable in the pH range of 9.0-12.0 at $25^{\circ}C$ for 36 h. and the half-life at $80^{\circ}C$ (pH 11.0) was 6 h. The enzyme was activated by $MgCl_2$ and inhibited by EDTA. With ${\rho}-nitrophenyl\;phosphate\;({\rho}NPP)$ as the substrate, the enzyme had a Michaelis constant $(K_m) $of $3.6{\times}10^{-5}M$, The enzyme preferentially hydrolyzed the phosphomonoester bond of AMP in ribonucleotides and glycerophosphate.

• The Korean Journal of Physiology
• /
• v.7 no.1
• /
• pp.1-11
• /
• 1973

To study the effects of Korean ginseng on mice visceral alkaline phosphatase activity and inorganic phosphate phosphorus level in the blood, the experimental groups of male and female mice were injected subcutaneously with 0.01 ml or 0.02 ml of Tyrode solution per 10 g of body weight daily, and 0. 1 mg or 0.2 mg of alcohol extract of ginseng which was diluted with Tyrode solution. After groups of mice were treated for 7 days or 14 days, the alkaline phosphatase activity in the jejunum, kidney and liver homogenate and serum were determined with sodium ${\beta}-glycerophophosphate$ as substrate, and quantified the content of inorganic phosphate phosphorus in the blood of above mice. 1. Alkaline phosphatase activity in the jejunum, kidney, liver and serum of male mice for the 7 day treatment with ginseng extract (0.1 mg/10 g) was increased by 15.28%, 12.86%, lg.05% and 11.70% respectively, compared with Tyrode solution group. 2. Alkaline phosphatase activity in the jejunum, kidney, liver and serum of female mice for the 7 day treatment with ginseng extract (0.2 mg/10 g) was increased 3.46%, 6.94%, 20.37% and 4.05 % respectively. 3. Alkaline phosphatase activity in the jejunum, kidney, liver and serum of male mice for the 14 day treatment with ginseng extract(0.1 mg/10 g) was increased·15.92%, 19.76f, 10.16% and -1.63 % respectively. 4. Alkaline phosphatase activity in the jejunum, kidney, liver and serum of female mice for the 14 day treatment with ginseng extract(0.2 mg/10 g) was increased 18.89%, 24.55%, 16.97% and 27.59% respectively. 5. Increasing activity of the enzyme in the liver of both male and female mice for the 7 day treatment was declined to some extent at the 14 day treatment, on the other hand, increasing activity of the enzyme in the jejunum, kidney and serum of mice for the 7 day treatment was more promoted at the 14 day treatment. 6. The 7 day and 14 day treatment with ginseng extract increased 20.20% and 20.96% of inorganic phosphate phosphorus in male blood, 22.38% and 17.57f in female blood respectively. In accordance with the results mutual relationships of ginseng, internal secretion, nucleic acid and alkaline phosphatase activity were disscussed.

• The Korean Journal of Zoology
• /
• v.31 no.4
• /
• pp.300-308
• /
• 1988

Human placental alkaline phosphatase (PLAP), one of the oncofetal antigen was purified from placentas through the procedures including butanol extraction, concanavalin A-Sephar-ose, DEAE-cellulose and Sephadex G-200 gel chromatography. Monoclonal antibodies (fibs) against human PMP were produced by hybridizing SP 210-Ag 14 mouse myeloma cells with spleen ceils of Balblc mice immunized with PLAP. Six stable monoclones uvere obtained by cloning tuvice in serial dilutions, and the monoclonal speclfidty of these MAbs was confirmed by biochemical and immunonogical criteria. Tumor marker의 하나인 태반형 alkaline phosphatase(PLAP)에 대한 단일 클론항체의 생산과 분석을 위하여, 태반조직을 재료로 butanol 추출법 및 concanavaline A-Sepharose, DEAE-cellulose, Sephadex G-200 gel 크로마토그라피법에 의하여 PLAP를 순수 분리하였다. 이를 항원으로 하여 하이브리도마 방법에 의해 항-PLAP 단일 클론항체를 생산 분비하는 안정된 6클론세포를 얻었으며 생화학적 및 면역학적 분석방법으로 이들의 단일 클론성을 확인하였다.

• Microbiology and Biotechnology Letters
• /
• v.23 no.2
• /
• pp.131-137
• /
• 1995

The alkaline phosphatase (K-ALPase) gene of Kluyveromyces fragilis has been cloned (1) and determined its base sequences (2) previously in our laboratory. When the K-ALPase gene was expressed in Escherichia coli and Saccharomyces cerevisiae, it showed a constitutive activity in E. coli, and a derepressed activity in S. cerevisiae in phosphate-limited medium. Northem hybridization experiment was performed to elucidate the transcription level of the K-ALPase gene. Northern experiment showed that transcription level of K-ALPase gene in S. cerevisiae was higher in phosphate depletion, but it was higher in high phosphate medium than in phosphate limited medium in K. fragilis. The transcription initiation site of the K-ALPase gene was determined by primer extension analysis. It matched nucleotide position - 169 in relation to the putative trnslational start site.

• The Journal of the Korean dental association
• /
• v.13 no.1
• /
• pp.37-45
• /
• 1975

• Journal of Periodontal and Implant Science
• /
• v.4 no.1
• /
• pp.12.2-12.2
• /
• 1974

• Korean Journal of Microbiology
• /
• v.23 no.3
• /
• pp.172-176
• /
• 1985

The present study was designed to investigate isoenzyme (ACPase, ALPase) pattern and its refulatory function between catabolically repressed and derepressed states in yeast, Saccharomyces uvarum. As the results, no other isoenzyme was detectable in acid phosphatase, but there were three isoenzyme types in aldaline phosphatase. Type "B" isoenzyme among alkaline phosphatases in catabolically repressed cell was derepressed, but in normally cultivated cell, type "C" isoenzyme was derepressed while type "B" activity was lowered. Type "B" isoenzyme could be postulated as repressible enzyme, type "A" as constityityve enzyme and type "C" as L-histidinol phosphatase, respectively, Also, it could be shown that type "B" ALPase, repressible enzyme, compensated for phosphate group supplier under catabolically repressed states. Protein profile in cytoplasmic soluble fraction of exponential phase cell was characterized by negative charged protein.

• Imaging Science in Dentistry
• /
• v.33 no.1
• /
• pp.51-57
• /
• 2003

Purpose: To investigate the effects of irradiation on the phenotypic expression of the MC3T3-El osteoblastic cell line, particularly an the expression of type I collagen and alkaline phosphatase mRNA. Materials and Methods: Cells were irradiated with a single dose of 0.5, 1, 2, 4, and 8 Gy at a dose rate of 5.38 Gy/min using a cesium 137 irradiator. The specimens were then harvested and RNA extraction was carried out at 1 and 3 days after irradiation. The extracted RNA strands were reverse-transcribed and the resulting cDNA fragments were amplified by PCR. Results: The irradiated cells demonstrated a dose-dependent increase in type I collagen mRNA expression relative to the control group, with a maximum level of type I collagen mRNA expression occurring at 8 Gy. The degree of type I collagen mRNA expression increased significantly at 1 day after irradiation, but little differences were found between the control group and at the 3rd day. The amount of alkaline phosphatase mRNA expression increased significantly at land 3 days after irradiation in the 1 Gy exposed group compared with the control group. Conclusion: The amount of type I collagen and alkaline phosphatase mRNA expression increased significantly 1 day after irradiation when compared with the control group.