• Journal of Microbiology
• /
• v.41 no.3
• /
• pp.202-206
• /
• 2003

A protein, exhibiting a high similarity to the major serine transporter of Escherichia coli, SstT, was found in Haemophilus influenzae Rd. A Na$\^$＋/-stimulated serine transport activity was also detected in the cells. The gene (sstT) for the Na$\^$＋//serine symporter from the chromosome of H. influenzae was cloned, and the properties of the transporter investigated. The serine transport activity was stimulated by Na$\^$＋/. The uptake of Na$\^$＋/ was elicited by the addition of serine or threonine into the cells, supporting the idea that these amino acids are transported by a mechanism of Na$\^$＋//substrate symport. No uptake of H$\^$＋/ was elicited by the influx of serine. The serine transport via the SstT of H. influenzae was inhibited by excess threonine, which was used as another substrate. The $K_{m}$ and the $V_{max}$ values for the serine transport were 2.5 ${\mu}$M and 14 nmol/min/mg protein, respectively.

• Microbiology and Biotechnology Letters
• /
• v.21 no.6
• /
• pp.577-581
• /
• 1993

The amino acid threonine was produced from glycine and ethanol in a reaction mixture using cell free extract of the methylotrophic bacterium isolated from soil and identified as mellthylo-bacterium sp. KJ29. Although the isolate could grow on carbon source other than methanol, only the cell free extract from the cells grown on methanol produced threonine. Methanol dehydrogenase (MDH) activity was present only in the cells grown on methanol when compared to the cells grown on heterotrophic substrates.

• Microbiology and Biotechnology Letters
• /
• v.35 no.2
• /
• pp.112-117
• /
• 2007

Protein kinases play very important role for maintaining viability in prokaryote and eukaryote. The metabolism of prokaryotic cell is generally regulated by bacterial two-component regulatory systems that are composed of histidine and asparitic acid kinases, however, some eukaryotic signal transduction system such as, serine and threonine kinases, have been also found to be involved in the regulation of morphogenesis and physiological differentiation in Streptomyces. Streptomyces griseus, a streptomycin producer, was expected to have varlous types of eukaryotic-type serine/threonine protein kinases, controlling morphogenesis. Thus, many steps of chromatographies were applied to isolate serine and threonine kinases from S. griseus IFO13350. The immunoaffinity steps using anti-phosphoserine, anti-phosphothreonine, and anti-phosphotyrosine agarose column chramatographies were successfully introduced to identify eukaryotic protein kinases from S. griseus IFO13350. Eight proteins with the expected molecular weight of 14, 29, 31, 35, 40, 52, 56, and 60 kDa, were identified on SDS-PAGE, and the their kination activity was confirmed by nonradioactive protein kination assay using FITC-labeled peptide as the substrate.

• The Journal of Internal Korean Medicine
• /
• v.13 no.2
• /
• pp.126-133
• /
• 1992

Comparative amino acid quantities on the blood analysis was carried out to investigate the amino acid specific characters on the blood in four type of physical constitution 1. In TAE-EUM-IN group, compared with control group, the proline and the serine were more observed. 2. In SO-EUM-IN group, compared with control group, the aspartic acid was more observed. 3. In SO-YANG-IN group, compared with control group, the proline was more observed. but the threonine and the aspartic acid were less observed. 4. In SO-EUM-IN group, compared with TAE-EUM-IN group, the aspartic acid and the serine were more observed. 5. In SO-YANG-IN group, compared with TAE-EUM-IN group, the serine and the proline were more observed, but the glutamic acid and the threonine were less observed. 6. In SO-YANG-IN group, compared with SO-EUM-IN group, the threonine and the aspartic acid were less observed.

• BMB Reports
• /
• v.36 no.4
• /
• pp.421-425
• /
• 2003

The serine/threonine protein kinase family is a large and diverse group of enzymes that are involved in the regulation of multiple cellular pathways. Elevated kinase activity has been implicated in many diseases and frequently targeted for the development of pharmacological inhibitors. Therefore, non-radioactive antibody-based kinase assays that allow high throughput screening of compound libraries have been developed. However, they require a generation of antibodies against the phosphorylated form of a specific substrate. We report here a time-resolved fluorescence assay platform that utilizes a commercially-available generic anti-phosphothreonine antibody and permits assaying kinases that are able to phosporylate threonin residues on protein substrates. Using this approach, we developed an assay for Cdc7/Dbf4 kinase activity, determined the $K_m$ for ATP, and identified rottlerin as a non-ATP competitive inhibitor of this enzyme.

• Korean Chemical Engineering Research
• /
• v.44 no.1
• /
• pp.97-105
• /
• 2006

In order to investigate the effect of inhibitors on L-threonine biosynthesis in Escherichia coli, we have constructed a metabolic network model of amino acid biosynthesis from L-aspartate to L-threonine by using available informations from literatures and databases. In the model, the effects of inhibitors on the biosynthesis of L-threonine was included as an appropriate mathematical form. For simulation study, we used initial values as L-aspartate 5 mM, ATP 5 mM, NADPH 2 mM, and observed the concentration changes of intermediate metabolites over concentration changes of respective inhibitors. As a result, we found that concentrations of intermediate metabolites were not significantly changed over concentration changes of L-lysine, L-methionine, and L-glutamate. But, there were considerable changes of intermediates over concentration changes of L-serine, L-cysteine, and L-threonine, which can be considered as essential effectors on L-threonine synthesis. Contrary, the synthesis of L-threonine seems to be not related to the amounts of L-aspartate, and inversely proportional to the accumulated amount of D,L-aspartic ${\beta}$-semialdehyde.

• YAKHAK HOEJI
• /
• v.34 no.6
• /
• pp.447-456
• /
• 1990

Various methods are available for the production of L-threonine. The microbial production of L-threonine has been achieved by breeding L-threonine analog-resistant auxotrophic mutants of various bacteria. The enzymatic production of L-threonine has been demonstrated by use of threonine metabolic enzymes such as threonine deaminase, threonine aldolase, or threonine dehydrogenase complex. Threonine synthesis from glycine and ethanol seems to be catalyzed by the enzymes Methanol dehydrogenase(MDH) and Serine hydroxymethyltransferase(SHMT), which was also found to catalyze the aldol condensation of glycine with acetaldehyde. The improved production of L-threonine has been achieved by amplifying the genes for the L-threonine biosynthetic enzymes using recombinant DNA techniques.

• BMB Reports
• /
• v.28 no.2
• /
• pp.124-128
• /
• 1995

Biodegradative threonine dehydratase purified from Serratia marcescens ATCC 25419 was inactivated by the arginine specific modification reagent, phenylglyoxal (PGO) and the lysine modification reagent, pyridoxal 5'-phosphate (PLP). The inactivation by PGO was protected by L-threonine and L-serine. The second order rate constant for the inactivation of the enzyme by PGO was calculated to be 136 $M^{-1}min^{-1}$. The reaction order with respect to PGO was 0.83. The inactivation of the enzyme by PGO was reversed upon addition of excess hydroxylamine. The inactivation of the enzyme by PLP was protected by L-threonine, L-serine, and a-aminobutyrate. The second order rate constant for the inactivation of the enzyme by PLP was 157 $M^{-1}min^{-1}$ and the order of reaction with respect to PLP was 1.0. The inactivation of the enzyme by PLP was reversed upon addition of excess acetic anhydride. Other chemical modification reagents such as N-ethylmaleimide, 5,5'-dithiobis (2-nitrobenzoate), iodoacetamide, sodium azide, phenylmethyl sulfonylfluoride and diethylpyrocarbonate had no effect on the enzyme activity. These results suggest that essential arginine and lysine residues may be located at or near the active site.

• Animal cells and systems
• /
• v.1 no.1
• /
• pp.135-142
• /
• 1997

Random sequencing of expressed sequence tags in roots of Chinese cabbage led to isolation of a partial cDNA clone, BR77, which encoded a putative protein kinase. Using the BR77 cDNA as a probe, we isolated a full-length cDNA encoding the Brassica campestris protein kinase 1 (Bcpk1). The Bcpt1 cDNA contained one open reading frame encoding a polypeptide of 439 amino acids. The putative polypeptide consisted of a short N-terminal region and a protein kinase catalytic domain. The catalytic domain of Bcpkl showed a high homology to cAMP- and calcium- phospholipid-dependent subfamilies of serine/threonine protein kineses. Eleven major catalytic domains in protein kineses were well conserved in Bcpk1. However, Bcpk1 contained a unique nonhomologous intervening sequence between subdomains VII and VIII, which was not found in protein kineses of animals and lower eukaryotes. Genomic DNA gel blot analysis showed that Bcpt1 genes might be present as three copies in the Chinese cabbage genome. These imply that Bcpk1 belongs to a plant-specific serine/threonine protein kinase subfamily.

• 한국생물공학회:학술대회논문집
• /
• 2003.04a
• /
• pp.543-543
• /
• 2003