• Journal of Microbiology
• /
• 제41권3호
• /
• 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.

• 한국미생물·생명공학회지
• /
• 제21권6호
• /
• 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.

• 한국미생물·생명공학회지
• /
• 제35권2호
• /
• pp.112-117
• /
• 2007

Protein kinase는 진핵생물과 원핵생물을 포함하는 모든 생명체에서 세포생존에 절대적으로 중요한 조절 기능을 담당한다. 일반적으로 원핵생물은 histidine 과 aspartic acid kinase로 구성된 bacterial two-component regulatory system에 의하여 환경변화에 따른 유전자의 발현이 조절되지만, 방선균을 비롯한 고등 원핵생물에서는 진핵생물성의 serine/threonine kinase들이 세포분화와 같은 분화과정을 조절하고 있다. Streptomycin 생산균인 Streptomyces griseus 균주에서도 다양한 serine/threonine kinase들이 존재하는 것으로 추정되며, 이들의 기능을 밝히는 것은 생명현상을 이해하는 중요한 열쇠를 제공해 줄 것으로 기대된다. 따라서, S. griseus로부터 protein kinase 를 동정하는 연구를 실시하였으며, 기존의 복잡한 chromatography법의 단점을 보완하기 위해 anti-phosphothreonine, anti-phosphoserine, anti-phosphotyrosine antibody를 이용한 immunoaffinity column chromatography 방법을 도입하였다. 실험 결과 약 14, 29, 31, 35, 40, 52, 56, 60 kDa의 단백질을 효과적으로 동정 할 수 있었으며, nonradioactive protein kination assay 방법으로 이들의 인산화능을 확인하였다.

• 대한한방내과학회지
• /
• 제13권2호
• /
• 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
• /
• 제36권4호
• /
• 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
• /
• 제44권1호
• /
• pp.97-105
• /
• 2006

본 연구에서는 대장균 내에서 L-threonine의 생합성에 영향을 미치는 저해제들에 대한 모사 연구를 위하여 L-aspartate에서 L-threonine까지의 아미노산 생합성 대사 네트워크를 문헌 및 데이터베이스를 통해 구축하였다. 또한 L-threonine 생합성에 영향을 미치는 저해제들을 수학적으로 모델링하여 효소 반응식에 적용시켰다. 모사 연구를 위해 초기 농도값을 L-aspartate 5 mM, ATP 5 mM, NADPH 2 mM으로 설정하고 저해제의 농도 변화에 따른 세포내 대사 물질들의 농도변화를 확인하였다. 그 결과 저해제 L-lysine, L-methionine, L-glutamate는 저해제 농도 변화에 따라 대사 물질들의 농도 변화가 없었다. 그러나 저해제 L-serine, L-cysteine 그리고 L-threonine의 경우 저해제의 농도 변화에 따라 세포내 대사물질들의 농도 곡선이 서로 다른 결과를 얻었다. 대장균 내에서 소비되어진 L-aspartate의 농도는 세포 내 생성되는 L-threonine과는 관련이 없었고, 생성되는 L-threonine의 농도는 세포 내에 축적된 D,L-aspartic ${\beta}$-semialdehyde에 반비례하였다.

• 약학회지
• /
• 제34권6호
• /
• 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
• /
• 제28권2호
• /
• 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
• /
• 제1권1호
• /
• 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
• /
• 한국생물공학회 2003년도 생물공학의 동향(XII)
• /
• pp.543-543
• /
• 2003