• Title/Summary/Keyword: serine

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Cloning of a Novel $Na^+$-Dependent L-Serine Specific Symporter Gene from Haemophilus influenzae Rd and Characteristics of the Transporter

  • Kim, Young-Mog;Rhee, In-Koo;Tsuchiya, Tomofusa
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.520-524
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    • 2004
  • A protein that exhibited a high similarity to a major serine transporter of Escherichia coli, SdaC, was found in Haemophilus injluenzae Rd. Also, $Na^+$-stimulated serine transport activity was detected in the cells. The sdaC of H. injluenzae was cloned and the properties of the transporter were investigated. The activity of serine transport was stimulated by $Na^+$. Uptake of $Na^+$ elicited by L-serine influx into cells was also observed, which supports the idea that L-serine is transported by a mechanism of $Na^+$serine symport. No uptake of $H^+$ elicited by L-serine influx was detected. This result was not consistent with that obtained with the homologous protein, SdaC of E. coli, which uses $H^+$as a coupling cation. The serine transport via the SdaC of H. influenzae was not inhibited by other amino acids such as threonine or D-serine like the SdaC of E. coli. Thus, the SdaC of H. influenzae is a $Na^+$-dependent L-serine specific symporter and an unusual natural mutant. The $K_m$ and the $V_{max}$, value for the serine transport in the SdaC of H. influenzae were $7.6\mu$M and 22.9 nmol/min/mg protein, respectively.

Characteristics of $Na^{+}$-dependent Serine Transport in Haemophilus Influenzae Rd

  • Kim, Young-Mog;Rhee, In-Koo;Park, Mi-Yeon;Chang, Dong-Suck;Tomofusa Tsuchiya
    • Journal of Microbiology
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    • v.41 no.2
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    • pp.78-82
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    • 2003
  • We identified two proteins in Haemophilus influenzae Rd that exhibited high similarity to two major serine transporters of Escherichia coli (SstT and SdaC). Then, we investigated serine transport in H. influenzae Rd and detected $Na^{+}$-stimulated L-serine transport activity. The optimum NaCl concentration for this stimulation was about 20 mM. The uptake of $Na^{+}$ by H. influenzae Rd was found to be elicited by L-serine influx, which supports the idea that L-serine is transported by a mechanism of $Na^{+}$/serine symport. No uptake of $H^{+}$ elicited by L-serine influx was detected. $Na^{+}$/serine symport activity was not inhibited by other amino acids such as L-threonine or D-serine. Two distinct Km values were obtained from the kinetic analysis of serine transport. Thus, two serine transport pathways may exist in H. influenzae Rd, and it appears that both systems are stimulated by $Na^{+}$.

Cloning of the Gene for Na$^{+}$/Serine-Threonine Symporter (sstT) from Haemophilus influenzae Rd and Characteristics of the Transporter

  • Kim, Young-Mog
    • Journal of Microbiology
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    • v.41 no.3
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    • pp.202-206
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    • 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.

Kinetic Characterization of an Iron-sulfur Containing Enzyme, L-serine Dehydratase from Mycobacterium tuberculosis H37Rv (Mycobacterium tuberculosis H37Rv로부터 유래된 철-황 함유 효소인 L-세린 탈수화효소의 동력학적 특성)

  • Han, Yu Jeong;Lee, Ki Seog
    • Journal of Life Science
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    • v.28 no.3
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    • pp.351-356
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    • 2018
  • L-Serine dehydratase (LSD) is an iron-sulfur containing enzyme that catalyzes the conversion of L-serine to pyruvate and ammonia. Among the bacterial amino acid dehydratases, it appears that only the L-serine specific enzymes utilize an iron-sulfur cluster at their catalytic site. Moreover, bacterial LSDs are classified into four types based on structural characteristics and domain arrangement. To date, only the LSD enzymes from a few bacterial strains have been studied, but more detailed investigations are required to understand the catalytic mechanism of various bacterial LSDs. In this study, LSD type II from Mycobacterium tuberculosis (MtLSD) H37Rv was expressed and purified to elucidate the biochemical and catalytic properties using the enzyme kinetic method. The L-serine saturation curve of MtLSD exhibited a typically sigmoid character, indicating an allosteric cooperativity. The values of $K_m$ and $k_{cat}$ were estimated to be $59.35{\pm}1.23mM$ and $18.12{\pm}0.20s^{-1}$, respectively. Moreover, the plot of initial velocity versus D-serine concentration at fixed L-serine concentrations showed a non-linear hyperbola decay shape and exhibited a competitive inhibition for D-serine with an apparent $K_i$ value of $30.46{\pm}5.93mM$ and with no change in the $k_{cat}$ value. These results provide insightful biochemical information regarding the catalytic properties and the substrate specificity of MtLSD.

Metagenome Resource for D-Serine Utilization in a DsdA-Disrupted Escherichia coli

  • Lim, Mi-Young;Lee, Hyo-Jeong;Kim, Pil
    • Journal of Microbiology and Biotechnology
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    • v.21 no.4
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    • pp.374-378
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    • 2011
  • To find alternative genetic resources for D-serine dehydratase (E.C. 4.3.1.18, dsdA) mediating the deamination of D-serine into pyruvate, metagenomic libraries were screened. The chromosomal dsdA gene of a wild-type Escherichia coli W3110 strain was disrupted by inserting the tetracycline resistance gene (tet), using double-crossover, for use as a screening host. The W3110 dsdA::tet strain was not able to grow in a medium containing D-serine as a sole carbon source, whereas wild-type W3110 and the complement W3110 dsdA::tet strain containing a dsdA-expression plasmid were able to grow. After introducing metagenome libraries into the screening host, a strain containing a 40-kb DNA fragment obtained from the metagenomic souce derived from a compost was selected based on its capability to grow on the agar plate containing D-serine as a sole carbon source. For identification of the genetic resource responsible for the D-serine degrading capability, transposon-${\mu}$ was randomly inserted into the 40-kb metagenome. Two strains that had lost their D-serine degrading ability were negatively selected, and the two 6-kb contigs responsible for the D-serine degrading capability were sequenced and deposited (GenBank code: HQ829474.1 and HQ829475.1). Therefore, new alternative genetic resources for D-serine dehydratase was found from the metagenomic resource, and the corresponding ORFs are discussed.

Extracellular Signal-regulated Kinase Activation Is Required for Serine 727 Phosphorylation of STAT3 in Schwann Cells in vitro and in vivo

  • Lee, Hyun-Kyoung;Jung, Jun-Yang;Lee, Sang-Hwa;Seo, Su-Yeong;Suh, Duk-Joon;Park, Hwan-Tae
    • The Korean Journal of Physiology and Pharmacology
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    • v.13 no.3
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    • pp.161-168
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    • 2009
  • In the peripheral nerves, injury-induced cytokines and growth factors perform critical functions in the activation of both the MEK/ERK and JAK/STAT3 pathways. In this study, we determined that nerve injury-induced ERK activation was temporally correlated with STAT3 phosphorylation at the serine 727 residue. In cultured Schwann cells, we noted that ERK activation is required for the serine phosphorylation of STAT3 by neuropoietic cytokine interleukin-6 (IL-6). Serine phosphorylated STAT3 by IL-6 was transported into Schwann cell nuclei, thereby indicating that ERK may regulate the transcriptional activity of STAT3 via the induction of serine phosphorylation of STAT3. Neuregulin-1 (NRG) also induced the serine phosphorylation of STAT3 in an ERK-dependent fashion. In contrast with the IL-6 response, serine phosphorylated STAT3 induced by NRG was not detected in the nucleus, thus indicating the non-nuclear function of serine phosphorylated STAT3 in response to NRG. Finally, we determined that the inhibition of ERK prevented injury-induced serine phosphorylation of STAT3 in an ex-vivo explants culture of the sciatic nerves. Collectively, the results of this study show that ERK may be an upstream kinase for the serine phosphorylation of STAT3 induced by multiple stimuli in Schwann cells after peripheral nerve injury.

Absorption of Carbon Dioxide into Aqueous Potassium Salt of Serine (Serine 칼륨염 수용액의 이산화탄소 흡수특성)

  • Song, Ho-Jun;Lee, Seung-Moon;Lee, Joon-Ho;Park, Jin-Won;Jang, Kyung-Ryong;Shim, Jae-Goo;Kim, Jun-Han
    • Journal of Korean Society of Environmental Engineers
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    • v.31 no.7
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    • pp.505-514
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    • 2009
  • Aqueous potassium salt of serine was proposed as an alternative $CO_2$ absorbent to monoethanolamine (MEA) and its $CO_2$ absorption characteristics were studied. The experiment has been conducted using screening test equipment with NDIR type gas analyzer and vapor-liquid equilibrium apparatus. $CO_2$ absorption/desorption rate and net amount of $CO_2$ absorbed in cyclic process are the criteria to assess the $CO_2$ absorption characteristics in this study. Effective $CO_2$ loading of potassium salt of serine and MEA are 0.425 and 0.230 respectively. Cyclic capacities are 0.354 and 0.298 for potassium salt of serine and MEA. The absorption rate of the potassium serinate decreased sharply at $CO_2$ loading is 0.1 and were maintained approximately at half of MEA. To enhance the absorption rate of aqueous potassium salt of serine, small quantities of rate promoters, namely piperazine and tetraethylenepentamine were blended, so that rich $CO_2$ loading were increased by 13.7% and 18.7% respectively. The rich $CO_2$ loading of potassium salt of serine was 29.2% and 35.0% higher than those of aqueous sodium and lithium salt of serine, respectively. The absorption rate of potassium salt of valine and isoleucine which have similar molecular structures to serine were lower than that of serine because of the presence of bulky side group. Precipitation phenomena during $CO_2$ absorption were discussed by the aid of literatures.

Combined Skin Moisturization of Liposomal Serine Incorporated in Hydrogels Prepared with Carbopol ETD 2020, Rhesperse RM 100 and Hyaluronic Acid

  • Kim, Hyeongmin;Ro, Jieun;Barua, Sonia;Hwang, Deuk Sun;Na, Seon-Jeong;Lee, Ho Sung;Jeong, Ji Hoon;Woo, Seulki;Kim, Hyewon;Hong, Bomi;Yun, Gyiae;Kim, Joong-Hark;Yoon, Young-Ho;Park, Myung-Gyu;Kim, Jia;Sohn, Uy Dong;Lee, Jaehwi
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.6
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    • pp.543-547
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    • 2015
  • We investigated the combined moisturizing effect of liposomal serine and a cosmeceutical base selected in this study. Serine is a major amino acid consisting of natural moisturizing factors and keratin, and the hydroxyl group of serine can actively interact with water molecules. Therefore, we hypothesized that serine efficiently delivered to the stratum corneum (SC) of the skin would enhance the moisturizing capability of the skin. We prepared four different cosmeceutical bases (hydrogel, oil-in-water (O/W) essence, O/W cream, and water-in-oil (W/O) cream); their moisturizing abilities were then assessed using a $Corneometer^{(R)}$. The hydrogel was selected as the optimum base for skin moisturization based on the area under the moisture content change-time curves (AUMCC) values used as a parameter for the water hold capacity of the skin. Liposomal serine prepared by a reverse-phase evaporation method was then incorporated in the hydrogel. The liposomal serine-incorporated hydrogel (serine level=1%) showed an approximately 1.62~1.77 times greater moisturizing effect on the skin than those of hydrogel, hydrogel with serine (1%), and hydrogel with blank liposome. However, the AUMCC values were not dependent on the level of serine in liposomal serine-loaded hydrogels. Together, the delivery of serine to the SC of the skin is a promising strategy for moisturizing the skin. This study is expected to be an important step in developing highly effective moisturizing cosmeceutical products.

Cloning and Expression of a Serine Proteinase Gene Fragment from Acanthamoeba culbertsoni

  • Park, Ki-Won;Kim, Tong-Soo;Na, Byoung-Kuk;Song, Chul-Yong
    • BMB Reports
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    • v.31 no.3
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    • pp.303-306
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    • 1998
  • Serine proteinase cDNA fragment from protozoan parasite Acanthamoeba culbertsoni was amplified by the reverse transcription-polymerase chain reaction (RTPCR) using degenerate oligonucleotide primers derived from conserved serine proteinase sequences. The amplified DNA fragment was subcloned and sequenced. The sequence analysis and alignment showed significant sequence similarity to other eukaryotic serine proteinases and conservation of the His, Asp, and Ser residues that form the catalytic triad. The cDNA fragment was cloned into the pGEMEX-1 expression vector and expressed in Escherichia coli. A resulting fusion protein of 56 kDa had proteolytic activity. The fusion protein reacted with sera of mice immunized with purified serine proteinase of A. culbertsoni in Western blot. Immune recognition of the fusion protein by mouse antisera suggested that the fusion protein may be valuable as a diagnostic reagent.

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효모 Pichia ciferrii mutant의 Tetraacetylphytosphingosine 생산에 미치는 아미노산의 영향

  • Hong, Seong-Gap;Yu, Yeon-U
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.205-207
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    • 2002
  • Experiments were carried out to the effect of various amino acids on the production of tetraacetylphytosphingosine (TAPS). Among various amino acids were tested, cysteine decreased the production of TAPS at 7 days of flask culture. Especially, Serine is precursor of TAPS but didn't affect the production of TAPS. Glycine and glutamate increased the production of TAPS. Especially, glutamate increased 20% of cell mass and 37% of TAPS production.

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