• Title/Summary/Keyword: D-aspartic acid

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Kinetic Study on the Enzymatic Production of D-Alanine from D-Aspartic Acid

  • Lee, Jae-Heung;Sung, Moon-Hee;Jeon, Yeong-Joong
    • Journal of Microbiology
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    • v.40 no.1
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    • pp.33-37
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    • 2002
  • An enzymatic reaction for the production of D-alanine from D-aspartic acid and pyruvate as substrates by a thermostable D-amino acid aminotransferase (D-AAT) was investigated at various conditions In the temperature range of 40-70$\^{C}$ and pH range of 6.0-9.5. The D-AAT was produced with recombinant E. coli BL21, which hosted the chimeric plasmid pTLK2 harboring the D-AAT from the novel thermophilic Bacillus sp. LK-2. The enzyme reaction was shown to follow the Ping Pong Bi Bi mechanism. The K$\_$m/ values for D-aspartic acid and pyruvate were 4.38 mar and 0.72 mM, respectively. It was observed that competitive inhibition by D-alanine, the product of this reaction, was evident with the inhibition constant K$\_$i/ value of 0.1 mM. A unique feature of this reaction scheme is that the decorboxylation of oxaloacetic acid, one of the products, spontaneously produces pyruvate. Therefore, only a catalytic amount of pyruvate is necessary for the enzyme conversion reaction to proceed. A typical time-course kinetic study skewed that D-alanine up to 88 mM could be produced from 100 mM of D-aspartic acid with a molar yield of 1.0.

Effects of 2,4-D and NAA on Nicotine Production during Callus Culture of Nicotiana gluauca Graham (담배(Nicotiana glauca Graham) Callus 배양시 Nicotine 생성에 미치는 2,4-D 및 NAA의 영향)

  • 여읍동
    • Journal of Plant Biology
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    • v.34 no.2
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    • pp.101-106
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    • 1991
  • Effects of two auxins, 2,4-dichlorophenoxyacetic acid (2,4-D) and a-napthaleneacetic acid (NAA) on nicotine production during callus culture of a wild tobacco (Nicotiana glauca) were investigated using a high performance liquid chromatography (HPLC). The high concentration ($11.5\;\mu\textrm{M}$)of 2,4-D and NAA had peaks of nicotine contents at 4th and 2nd week, respectively. Thereafter, the concents decreased and the nicotine was metabolized to other alkaloids. The low concentration ($1.5\;\mu\textrm{M}$) of 2,4-D on the medium supplemented with 0.1 mM of L-aspartic acid or L-arginine inhibited nicotine production. However, the low NAA promoted it only when the medium was supplemented with L-aspartic acid. From these results, it could be concluded that both auxins exhibit different action mechanisms on nicotine production pathway and the low NAA promotes the activities for the pathway with L-aspartic acid as a precursor.cursor.

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Combination Effect of ʟ-Arginine and ʟ-Aspartic acid on Saltiness Enhancement of NaCl Solution (ʟ-아르기닌과 ʟ-아스파라긴산 혼합이 NaCl 짠맛 향상에 미치는 영향)

  • Kim, Yong-Duk;Park, Jung-Ha;Park, Bok-Jun;In, Man-Jin;Park, Dong-Chel;Oh, Nam-Soon
    • Journal of Applied Biological Chemistry
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    • v.57 no.3
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    • pp.251-254
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    • 2014
  • In order to investigate the combination effect of $\small{L}$-arginine and $\small{L}$-aspartic acid on salt enhancement, the saltiness and bitterness of various mixtures of $\small{L}$-arginine and $\small{L}$-aspartic acid were evaluated using the electronic tongue and sensory tests. Increasing the molar ration of $\small{L}$-arginine against $\small{L}$-aspartic acid enhanced the salty taste of NaCl, whereas increasing the molar ration of $\small{L}$-aspartic acid against $\small{L}$-arginine significantly suppressed the bitter taste of $\small{L}$-arginine. Therefore, combination of $\small{L}$-arginine and $\small{L}$-aspartic acid can be utilized as a saltiness enhancer and its suitable combination ratio was showed as $\small{L}$-arginine : $\small{L}$-aspartic acid = 1.00:0.98-1.00 on basis of molar concentration.

Development of a UPLC-MS/MS method for the therapeutic monitoring of L-asparaginase

  • Jeong, Hyeon-Cheol;Kim, Therasa;Yang, Deok-Hwan;Shin, Kwang-Hee
    • Translational and Clinical Pharmacology
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    • v.26 no.3
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    • pp.134-140
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    • 2018
  • This study aimed to develop a UPLC-MS/MS method for determining plasma levels of L-aspartic acid and L-asparagine and the activity of L-asparaginase. L-aspartic acid, L-asparagine, and L-aspartic acid-2,3,3-$d_3$ were extracted from human plasma by protein precipitation with sulfosalicylic acid (30%, v/v). The plasma samples were analyzed using an Imtakt Intrada amino acid analysis column with 25 mM ammonium formate and 0.5% formic acid in acetonitrile as the mobile phase with step gradient method at a flow rate of 0.5 mL/min. The injection volume was $5{\mu}L$, and the total run time was 15 min. Inter- and intra-batch accuracies (%) ranged from 96.62-106.0% for L-aspartic acid and 89.85-104.8%, for L-asparagine, and the coefficient of variation (CV%) did not exceed 7%. The validation results for L-aspartic acid and L-asparagine satisfied the specified criterion, however, the results for L-asparaginase activity assay showed a borderline validity. This study could be a foundation for further development of therapeutic drug monitoring systems using UPLC-MS/MS.

Total Synthesis of Sodium (3S,4R)-3-[2-(2-Aminothiazol-4-yl)-(Z)-2-methoxyiminoacetamido]-4-methoxymethyl-2-azetidinone-1-sulfonate from D-Aspartic Acid

  • Chung Bong Young;Kim, Jin Yeon;Nah Cha Soo;Han Kee Jong;Park, Jong Ok
    • Bulletin of the Korean Chemical Society
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    • v.13 no.3
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    • pp.315-316
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    • 1992
  • Sodium (3S,4R)-3-[2-(2-aminothiazol-4-yl)-(Z)-2-methoxyi minoacetamido]-4-methoxymethyl-2-azetidinone-1- sulfonate (2) was synthesized in fourteen steps from D-aspartic acid. Starting from D-aspartic acid, (3S,4R)-3-amino-1-t-butyldimethylsilyl-4-methoxym ethyl-2-azetidinone (12) was synthesized in ten steps. Acylation of the amino group of 12 with $2-amino-{\alpha}-(methoxyimino)-4-thiazoleacetic$ acid, desilylation, sulfonation, and ion exchange afforded sodium (3S,4R)-3-[2-(2-aminothiazol-4-yl)-(Z)-2-methoxyi minoacetamido]-4-methoxymethyl-2-azetidinone-1- sulfonate (2). This new ${\beta}-lactam$ compound 2 showed low antibacterial activities.

The Study of Age Estimation from Tooth using the Racemization of Aminoacid (아미노산의 라세미화 반응을 이용한 치아로부터의 연령감정에 관한 연구)

  • Hee-Kyung Kim;Chong-Youl Kim
    • Journal of Oral Medicine and Pain
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    • v.14 no.1
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    • pp.43-55
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    • 1989
  • The need of age estimation for identification was increased by complexity of society, and the tooth was used widely for age estimation because of less individual deviation than the other organ. The age estimation using the tooth had several methods. Recently, the one using the racemization of aminoacid in the tooth was admitted more accurate than the other methods, especially in old age. But, this study was not tried in our country, and I would report the result of experiment about age estimation using racemization of dentine. I selected 40-Whole dentine sample from extracted teeth, those were reserved in natural dried condition for 2 weeks~ 1year and calculated the estimation of age from the ratio of D-aminoacid and L-aminoacid (D/L ratio) using gaschromatography and the results were below. 1. The aminoacids showed apparent K/L ratio in dentine were aspartic acid, serine. 2. The aspartic acid showed the highest racemic rate and its rate was 0.0012$\pm$0.0003/yr. 3. The relation between the actual age and K/L ratio was very positive correlation(r+0.954) in the estimation of age using aspartic acid. 4. The deviation between the estimated age using D/L ratio of aspartic acid and actual age was $\pm$3.32.

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D99 Type I Signal Peptidase Implicated Stabilizing the Protein Structure (Type I 신호펩디드 가수분해효소에 존재하는 D99 아미노산 잔기의 구조적 역할 가능성)

  • Sung, Meesook;Eunyoung Han;Lee, Hoyoung
    • Microbiology and Biotechnology Letters
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    • v.31 no.2
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    • pp.140-144
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    • 2003
  • Type Ⅰ signal peptidase is an integral membrane protein that functions to cleave signal peptides from secreted and membrane proteins. The enzyme serves as a potential target for the development of novel antibacterial agents due to its unique physiological properties. Despite being one of the best characterized enzymes, the catalysis of Type Ⅰ signal peptidase still remains controversy over the catalytic serine/lysine dyad mechanism. It appears that the dyad proteases are generally less efficient than the prototypical serine/histidine/aspartic acid triad found in most enzymes, although Type Ⅰ signal peptidase is an exception to this rule. In this paper, we have proposed that Type Ⅰ signal peptidase may act as the serine/lysine/aspartic acid triad cataltytic mechanism. Therefore, the aspartic acid 99 residue in the E. coli signal peptidase was chosen and mutated to an alanine to see if there is any possible role of the aspartic acid in the catalytic function. Type Ⅰ signal peptidase D99A protein was inactive in vitro assay using the procoat synthesized by in vitro transcription translation. However, the mutant was active using a highly sensitive in vivo assay. Pulse-chase experiments show that the replacement of aspartic acid 99 with alanine results in a very unstable signal peptidase molecule. Therefore, we conclude that it is unlikely that the residue is directly involved in catalysis, but rather plays an important role in stabilizing the protein structure.

Bioavailability of Aspartic Acid Chelated Calcium in Calcium Deficient Rats (아스파르트산 킬레이트 칼슘의 칼슘 결핍쥐에서의 생물학적 유용성)

  • Park, Myoung-Gyu;Ha, Tae-Yul;Shin, Kwang-Soon
    • Journal of Nutrition and Health
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    • v.44 no.6
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    • pp.474-480
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    • 2011
  • Calcium (Ca) is an essential element to maintain body homeostasis. However, many factors disturb calcium absorption. Aspartic acid chelated calcium (AAC) was synthesized by new methods using calcium carbonate and aspartic acid. This study was carried out to investigate the bioavailability of AAC in Ca-deficient rats. The experimental groups were as follows: NC; normal diet control group, CD-C; untreated control group of Ca-deficient (CD) rats, CD-$CaCO_3$; $CaCO_3$ treated group of CD rats, CD-AAC; AAC treated group of CD rats, and CD-SWC; and seaweed-derived Ca treated group of CD rats. The Ca content of various types of Ca was held constant at 32 mg/day, and the four CD groups were fed for 7 days after randomized grouping. Ca content in serum, urine, and feces within feeding periods were analyzed to confirm Ca absorption. Serum Ca content was significantly higher in the CD-AAC (11.24 mg/dL) and CD-SWC (10.12 mg/dL) groups than that in the CD-C (8.6 mg/dL) group 2 hours following the first administration. The Ca content in feces was significantly lower in the CD-AAC (35.4 mg/3 days) and CD-SWC (71.1 mg/3 day) groups than that in the CD-$CaCO_3$ (98.7 mg/3 days) group (p > 0.05). AAC had a 2.3-fold higher absorption rate of Ca than that of SWC. No differences in fibula length were observed in the NC and CD groups. The fibula weights of the CD-AAC (0.33 g) and CD-SWC (0.33 g) groups increased compared to those in the CD-C (0.27 g) group; however, no significant difference was observed between the CD groups. We conclude that bioavailability of AAC is higher than that of seaweed-derived Ca or inorganic Ca. Thus, these findings suggest the AAC has potential as a functional food material related to Ca metabolism.

Nanoliposomes of L-lysine-conjugated poly(aspartic acid) Increase the Generation and Function of Bone Marrowderived Dendritic Cells

  • Im, Sun-A;Kim, Ki-Hyang;Ji, Hong-Geun;Yu, Hyoung-Gyoung;Park, Sun-Ki;Lee, Chong-Kil
    • IMMUNE NETWORK
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    • v.11 no.5
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    • pp.281-287
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    • 2011
  • Background: Biodegradable polymers have increasingly been recognized for various biological applications in recent years. Here we examined the immunostimulatory activities of the novel poly(aspartic acid) conjugated with L-lysine (PLA). Methods: PLA was synthesized by conjugating L-lysine to aspartic acid polymer. PLA-nanoliposomes (PLA-NLs) were prepared from PLA using a microfluidizer. The immunostimulatory activities of PLA-NLs were examined in mouse bone marrow-derived dendritic cells (BM-DCs). Results: PLA-NLs increased the number of BM-DCs when added to cultures of GM-CSF-induced DC generation on day 4 after the initiation of cultures. Examination of the phenotypic properties showed that BM-DCs generated in the presence of PLA-NLs are more mature in terms of the expression of MHC class II molecules and major co-stimulatory molecules than BM-DCs generated in the absence of PLA-NLs. In addition, the BM-DCs exhibited enhanced capability to produce cytokines, such as IL-6, IL-12, TNF-${\alpha}$ and IL-$1{\beta}$. Allogeneic mixed lymphocyte reactions also confirmed that the BMDCs were more stimulatory on allogeneic T cells. PLA- NL also induced further growth of immature BM-DCs that were harvested on day 8. Conclusion: These results show that PLA-NLs induce the generation and functional activities of BM-DCs, and suggest that PLA-NLs could be immunostimulating agents that target DCs.

Free amino acids of brown rice in relation to specific gravity grade (비중선별(比重選別) 현미중(玄米中) 유리 Amino산 함량)

  • Park, H.;Chun, J.K.;Cho, I.H.
    • Applied Biological Chemistry
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    • v.15 no.1
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    • pp.35-40
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    • 1972
  • The contents of free amino acids in deembryod brown rice of two varieties were investigated by amino acid autoanalizer in relation to specific gravity grade. The analytical methods of free amino acid were also discussed. 1) The lower the specific gravity of the unhulled rice the higher the content of total free amino acids in the deembryod brown rice, and the similar trend appears to hold on each amino acids. 2) Main free amino acids were serine+asparagine, glutamic acid, aspartic acid, alanine and valine, and maximum values of them were 7.3, 5.1, 4.0, 3.4, 0.9mg/100g rice, respectively. They consist about 85% of total free amino acids in most cases. 3) The contents of soluble nitrogen and free amino acids appear to be lower in high protein variety (IR 667) than in low protein variety (Jinhung). The percentage of free amino acid nitrogen to soluble nitrogen, however, appears to be higher in high protein variety (IR 667). 4) Alanine was much lower than aspartic acid in IR 667 having Indica blood while alanine appears to be higher than aspartic acid in Jinhung (Japonica rice) suggesting varietal difference in amino acid metabolism. 5) Threonine peak was overlaped with glutamine, and serine was with asparagine in this study.

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