• Journal of the Korean Society of Marine Environment & Safety
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• v.23 no.7
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• pp.885-892
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• 2017

We estimated the limiting nutrient and DIP limiting history based on alkaline phosphatase (APase) activity during the spring of 2017 in the Southern Sea, Korea. In the frontal area, concentration of dissolved inorganic phosphorus (DIP), dissolved inorganic nitrogen (DIN): DIP ratio and Chlorophyll a (Chl-a) were < $0.2{\mu}M$, 23.2 and $2.2{\mu}g/L$, respectively, indicating high productivity despite DIP limiting. The relationship between APase and DIP indicates that the study area had limited DIP because of a strongly reverse correlation (r= -0.81; P<0.001). Relationship between APase and Chl-a (r=0.61, p<0.001) also indicated that APase may have been induced by phytoplankton (ca. 60 %) and bacteria (ca. 40 %). In DIP limiting history in this study area, frontal area and non-frontal areas might have induced long-term DIP limitation and the recent relief from DIP-limitation, respectively, based on distributions of dissolved APase and particulate APase. Thus, these results suggest that by measuring the enzyme that hydrolyzes organic matter such as APase in frontal area, it is possible to estimate temporal and spatial characteristics of limiting nutrient, thereby improving our understanding of biogeochemistry cycles.

• Journal of Microbiology and Biotechnology
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• v.7 no.2
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• pp.127-131
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• 1997

Alkaline phosphatase (APase) was found to be inducible in Anabaena sp. strain CA Growth was less than control in presence of most amino acids except glycine and serine, but most amino acids enhanced APase activity. Highest APase activity was recorded in tyrosine supplemented culture followed by hydroxyproline, cystein, valine and glutamic acid. Threonine supplemented material showed lowest APase level (1.8 nmol/mg protein/min). Lactose, glucose, sodium pyruvate and succinate stimulated growth but not APase activity. APase activity was high in the presence of sucrose, mellibiose, mannitol, arabinose, maltose and sorbose, even though the growth in these supplements was less than in control. Organic phosphate sources supported good growth of the organism. Best growth occurred in presence of inorganic phosphate, adenosine diphosphate, fructose 1,6-diphosphate or ribulose 1,5-diphosphate, followed by other phosphorus sources tested. APase activity in presence of any of the organic phosphate sources was 3 to 5 fold low as compared to phosphate limited culture. Also, there was no APase activity in cultures grown on inorganic phosphate. These data indicate that most amino acids and a few carbohydrates (sucrose, mellibiose, arabinose and sorbose) are suitable for APase production. Lactose, glucose, pyruvate or succinate may be used as a carbon source during photoheterotrophic growth of the cyanobacterium. Glycine and serine are preferred nitrogen sources for its growth. Phosphate repressible APase activity has been found in Anabaena sp. strain CA.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.43 no.5
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• pp.540-546
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• 2010

We investigated variations in alkaline phosphatase (APase) activity and alkaline phosphatase hydrolyzable phosphorus (APHP) in northern Gamak Bay from September to December 2009. Dissolved inorganic nitrogen (DIN) and dissolved inorganic phosphorus (DIP) decreased gradually, and the DIN/DIP ratio was higher than the Redfield ratio (16) based on molecular concentrations during most of the observation period. The total APase (T-APase) activity increased with decreasing DIP concentration; i.e., the Relationship between T-APase and DIP showed a high negative correlation (r=-0.80, P<0.001), with APase activity being a good indicator of DIP limiting the Redfield ratio. The T-APase was positively correlated with the concentration of chlorophyll a (r=0.73, P<0.001). This suggests that a major portion of APase activity in northen Gamak Bay seawater is attributed to phytoplankton. The proportion of APHP among dissolved organic phosphorus (DOP) was low in September and high in November. Thus, APase-producing phytoplankton may be able to grow by utilizing APHP as a phosphorus source in autumn when DIP is limiting. Thus, APase activity and the use of DOP by phytoplankton may play an important role in the growth of phytoplankton under DIP limiting conditions such as those of northern Gamak Bay.

• Journal of The Korean Society of Grassland and Forage Science
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• v.17 no.3
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• pp.285-292
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• 1997

This study was conducted to obtain the transgenic Brnssica napus plants with tobacco Apase gene using the binary vector system of Agrobacteriurn fumefociens. The results obtained were summarized as follows: A repressible acid phosphatase gene of Saccharon~yces cerevisiae, pho105 was used for screening of tobacco Apase cDNA. In order to identify Apase gene in tobacco genome, Southern blot analysis was pcrformed and the Apase gcnc may be present as a single copy, or at most two or three copies, in tobacco genome. To isolate the tobacco Apase gene, tobacco cDNA library was constructed using purifed mRNA from -Pi treated tobacco root and the plaque forming unit of the library was 2.8 x $10^5$ pfu/m${\ell}$, therefore the library might cover all expressed mRNAs. Using pho5 as a probe. tobacco Apase cDNA was cloned, and restriction mapping and Southern blot analysis of cDNA insert were revealed that the 3.6 kb cDNA contained tobacco acid phosphatase cDNA. Plasmid pGA695 -tcAPl was constructed by subcloning tobacco Apase cDNA into the Hind site of pGA695 with 35s promoter which can be expressed constitutively in plants. The Brassica napus cotyledonary petioles were cocultivated with the ,4 grobacteriunz and transferred to the selection medium. The transformed and regenerated plants were transplanted to soil medium. Southern blot analysis was done on the transformed plants, and it was confirmed that a foregin gene was stably integrated into the genonies of B. nnpus plants.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.15 no.1
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• pp.16-24
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• 2010

Utilization of dissolved organic phosphorus (DOP) and alkaline phosphatase (APase) activity by Skeletonema costatum, Chaetoceros didymus, Alexandrium tamarense and Heterosigma akashiwo under the phosphorus deficient condition were examined in the laboratory. S. costatum, C. didymus, A. tamarense and H. akashiwo could make use of phosphomonoester and nucleotide compounds for the growth of them as a phosphorus source. APase activity of S. costatum, C. didymus, A. tamarense and H. akashiwo began to be activated at dissolved inorganic phosphorus (DIP) concentrations less than $0.30\;{\mu}M$, $0.33\;{\mu}M$, $2.04\;{\mu}M$ and $0.63\;{\mu}M$ respectively, and their maximum APase activity were $0.01\;pmol\;cell^{-1}\;hr^{-1}$, $0.11\;pmol\;cell^{-1}\;hr^{-1}$, $1.63\;pmol\;cell^{-1}\;hr^{-1}$ and $0.19\;pmol\;cell^{-1}\;hr^{-1}$, respectively. Although each phytoplankton species displayed different APase activity for DOP utilization, their maximum APase activities were higher than maximum phosphorus uptake rates, inferring that these species might be able to keep growing under DIP-limited conditions thought utilizing effectively the hydrolized product of DOP. This result also implies that utilization of DOP might contribute to not only the growth of red tide forming phytoplankton but also the interspecific competition among phytoplankton species in coastal environments.

• BMB Reports
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• v.36 no.6
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• pp.597-602
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• 2003

Acid phosphatases (APases) play a role in the release of phosphate in organic complexes in soil. We investigated tissue- and isoform-specific responses of APases to phosphorus (P) deficiency in three rice genotypes; Dasan-byeo, Sobi-byeo, and Palawan. The levels of shoot APase activity per protein were similar in the three genotypes. They significantly decreased with P deprivation that was longer than seven days. Root APase activity per protein was two- to three-fold higher in Dasan than in Sobi and Palawan. In all genotypes the APase activity increased in P-deficient plants, but the increase was higher in Sobi and Palawan. After 21 days of P deprivation, secreted APase activity increased more than eight-fold in Dasan and two-fold in Sobi and Palawan. Isoform profiles of shoot and root APases were most diverse in Dasan. The activities of the major isoforms in P-deficient shoots decreased in all three genotypes. Depending on the genotypes, further increases in constitutive isoforms and new induction of one to four isoforms occurred in P-deficient roots. The results indicate that tissue and genotype differences in the response of APase to P deficiency are primarily facilitated by the different responses of the isoforms.

• Microbiology and Biotechnology Letters
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• v.19 no.1
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• pp.37-44
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• 1991

- I aimed to isolate trans-acting factors involved in the basal expression level of eukaryotic genes. One of the yeast histidine biosynthetic gene, HIS5 was taken as a model for this study. HIS5 gene has a substantial basal level in amino acid rich medium and is derepressed if starved for any single amino acid. The derepression is mediated by cis-acting DNA sequences 5'-TGACTC-3' found in 5' non-transcribed region of the gene and trans-acting factors including GCN4 as positive factor and its negative factor GCDI 7, and GCNZ as a negative factor of GCD17. I first investigated the role of these trans-acting factors in HIS5 basal expression level by using HIS5-pH05 fusion in which expression of pH05 gene encoding inorganic phosphate-repressible acid phosphatase (APase) is regulated by HIS5 promoter. Strain with gcn2 or gcn4 mutation showed 3 to 4 fold lower APase activity than wild type. The level of APase activity was similar in gcn2 and gcn4 mutants. Trans-acting factors involved in basal level were identified by isolating 14 mutants showing increased expression of HISSPH05 fusion from gcn4 background. All the mutants carry a single nuclear recessive mutation and fall into four complementation groups, designated as bell (basal expression level), be12, be23 and be14.

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

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.322.1-322.1
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• 2002

Lawsone methyl ether (LME. 2-methoxy-1, 4-naphthoquinone) is a natural compound found in balsaminaceae. In this study the effect of LME on the release of renal dipeptidase (RDPase) and alkaline phosphatase (APase) known as glycosylphosphatidylinositol (GPI) anchored proteins was examined from the renal proximal tubules. Compared with control, LME (0.5mM) increased RDPase release (218%) and APase release (135%). The increase of RDPase release by LME showed concentration-dependent effect but the release pattern of APase did not. (omitted)

• Journal of Microbiology and Biotechnology
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• v.16 no.2
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• pp.272-279
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• 2006

The gene encoding Thermus sp. T351 alkaline phosphatase (T351 APase) was cloned and sequenced. The gene consisted of 1,503 bp coding for a protein with 500 amino acid residues including a signal peptide. The deduced amino acid sequence of T351 APase showed relatively low similarity to other Thermus APases. The T351 APase gene was expressed under the control of the T7lac promoter on the expression vector pET-22b(+) in Escherichia coli BL21 (DE3). The expressed enzyme was purified by heat treatment, and $UNO^{TM}$ Q and $HiTrap^{TM}$ Heparin HP column chromatographies. The purified enzyme exhibited high activity at extremely alkaline pHs, reaching a maximum at pH 12.0. The optimum temperature of the enzyme was $80^{\circ}C$, and the half-life at $85^{\circ}C$ was approximately 103 min. The enzyme activity was found to be dependent on metal ions: the addition of $Mg^{2+}$ and $CO^{2+}$ increased the activity, whereas EDTA inhibited it. With p-nitrophenyl phosphate as the substrate, T351 APase had a Michaelis constant ($K_{m}$) of $3.9{\times}10^{-5}M$. The enzyme catalyzed the hydrolysis of a wide variety of phosphorylated compounds.