• Title/Summary/Keyword: L-tryptophan

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Cell Biological Studies of the EfEect of Serotonin on Chick Embryogenesis (초기계배 발생에 미치는 Serotonin의 영향에 관한 세포생물학적 연구)

  • 최임순;주충노
    • The Korean Journal of Zoology
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    • v.30 no.4
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    • pp.432-444
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    • 1987
  • Very early stage of chick embryo cultivated in the medium containing DLtryptophan by beaker method has been investigated in vitro morphologically using electron microscopy at cellular level and found that the development of tryptophan treated chick embryos corresponding to 18∼66 hrs incubation was impaired and york granule degradation was significantly delayed. It was also found that DNA, RNA and protein biosynthesis of tryptophan treated chick embryo was greatly lowered than those of control group. Conversion of L-tryptophan into serotonin was traced using 14C-L-tryptophan and found that 13.8cA of added radioactivity was recovered from serotonin formed during 18 hrs incubation and the amounts of serotonin formed were depend upon added amount of tryptophan in e99 yolk. It seemed that the serotonin formed from external tryptophan might inhibit the degradation of yolk granule by feedback mechanism, resulting in malformation of chick embryogenesis.

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A Putative Peptide Synthetase from Bacillus subtilis 713 Recognizing $_{L}-Lysine,{\;}_{L}-Tryptophan,{\;}and{\;}_{L}-Glutamic$ Acid

  • Kim, Kyoung-Rok;Lee, In-Hyung;Suh, Joo-Won
    • Journal of Microbiology and Biotechnology
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    • v.11 no.5
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    • pp.798-803
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    • 2001
  • Peptide synthetases produced from various microorganisms are multifunctional enzyme complexes and their substrates are recognized and activated by adenylation domains. To identify the substrate specificity of the peptide synthetase isolated from Bacillus subtilis 713, known to produce an antifungal peptide, two adenylation domains containing the minimal functional portion were expressed and purified. ATP-ppi exchange experiments and kinetic studies revealed that the two adenylation enzymes had a substrate specificity to $_{L}-lysine{\;}and{\;}_{L}-tryptophan$, respectively. In addition, based on a signature sequence comparison, the substrate of the third domain was predicted to be L-glutamic acid. These results suggest that this peptide synthetase is novel because there has been no previous report on a peptide synthetase that uses $_{L}-lysine,{\;}_{L}-tryptophan,{\;}and{\;}_{L}-glutamic$ acid as substrates in that order.

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Controlled Fed-Batch Cultivation of Escherichia coli Mutant for L-Tryptophan Production (대장균 변이주의 조절식 유가배양법에 의한 L-트립토판 생산)

  • Lee, In-Young;Kim, Myung-Kuk;Kho, Yung-Hee;Kwak, Moo-Young;Lee, Hosull;Lee, Sun-Bok
    • Microbiology and Biotechnology Letters
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    • v.16 no.6
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    • pp.450-456
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    • 1988
  • For optimal production of L-tryptophan using a regulatory mutant of Escherichia coli the relationship between product formation and acid production was investigated. Experimental results showed that the production level of L-tryptophan was lowered as the specific acid production rate increased. In order to reduce the amount of acid produced during the fermentation, a controlled fed-batch fermentation was employed. In this fed-batch process, the feed rate of the nutrient feed medium was controlled in relation to the oxygen level in the culture and thus the growth of the cells was regulated in such n way that the oxygen demand of the culture could not exceed the oxygen sup-ply. When E. coli cells were cultivated in a controlled fed-batch mode of tormentor operation, the specific acid production rate was significantly reduced and L-tryptophan production was increased as much as five times that obtained in a conventional fed-batch fermentation.

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Electrochemical Investigation of Tryptophan at a Poly(p-aminobenzene sulfonic acid) Film Modified Glassy Carbon Electrode

  • Ya, Yu;Luo, Dengbai;Zhan, Guoqin;Li, Chunya
    • Bulletin of the Korean Chemical Society
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    • v.29 no.5
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    • pp.928-932
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    • 2008
  • A glassy carbon electrode (GCE) modified with poly(p-aminobenzene sulfonic acid) [Poly(p-ABSA)] film is fabricated by voltammetric technique in phosphate buffer solution (pH 8.0) containing $5.0\;{\times}\;10^{-3}\;mol\;L^{-1}$p- ABSA. Electrochemical behaviors of tryptophan at the Poly(p-ABSA) film electrode are investigated with voltammetry. The results indicate that the electrochemical response of tryptophan is improved significantly in the presence of poly(p-ABSA) film. Compared with the bare glassy carbon electrode, the Poly(p-ABSA) film electrode remarkably enhances the irreversible oxidation peak current of tryptophan. Some parameters such as voltammetric sweeping segments for the electrochemical polymerization, pH, accumulation potential and accumulation time are optimized. Under the optimal conditions, the oxidation peak current is proportional to tryptophan concentration in the range of $1.0\;{\times}\;10^{-7}$ to $1.0\;{\times}\;10^{-6}\;mol\;L^{-1}$, and $2.0\;{\times}\;10^{-6}$ to $1.0\;{\times}\;10^{-5}\;mol\;L^{-1}$ with a detection limit of $7.0\;{\times}\;10^{-8}\;mol\;L^{-1}$. The proposed procedure is successfully applied to the determination of tryptophan in a commercial amino acid oral solution.

Effects of tktA, $aroF^{FBR}$and aroL Expression in the Tryptophan-Producing Escherichia coli

  • Kim, Tae-Hyun;Namgoong, Suk;Kwak, Joon-Hyeok;Lee, Se-Yong;Lee, Heung-Shick
    • Journal of Microbiology and Biotechnology
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    • v.10 no.6
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    • pp.789-796
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    • 2000
  • In order to analyze the effects of tktA, $aroF^{FBR}$, and aroL expression in a tryptophan-producing Escherichia coli, a series of plasmids carrying the genes were constructed. Introduction of tktA, $aroF^{FBR}$, and aroL into the E. coli strain resulted in approximately 10-20 fold increase in the activities of transketolase, the feedback inhibition-resistant 3-deoxy-D-arabinoheptulsonate-7-phosphate synthase, and shikimate kinase. Expression of $aroF^{FBR}$ in the aroB mutant strain of E. coli resulted in the accumulation of 10 mM of 3-deoxy-D-arabinoheptulsonate-7-phosphate (DAHP) in the medium. Simultaneous expression of tktA and $aroF^{FBR}$ in the strain further increased the amount of excreted DAHP to 20 mM. In contrast, the mutant strain which has no gene introduced accumulated 0.5 mM of DAHP. However, the expression of tktA and $aroF^{FBR}$ in a tryptophan-producing E. coli strain did not lead to the increased production of tryptophan, but instead, a significant amount of shikimate, which is an intermediate in the tryptophan biosynthetic pathway, was excreted to the growth medium. Despite the fact that additional expression of shikimate kinase in the strain could possibly remove 90% of excreted shikimate to 0.1 mM, the amount of tryptophan produced was still unchanged. Removing shikimate using a cloned aroL gene caused the excretion of glutamate, which suggests disturbed central carbon metabolism. However, when cultivated in a complex medium, the strain expressing tktA, $aroF^{FBR}$, and aroL produced more tryptophan than the parental strain. These data indicate that additional rate-limiting steps are present in the tryptophan biosynthetic pathway, and the carbon flow to the terminal pathway is strictly regulated. Expressing tktA in E. coli cells appeared to impose a great metabolic burden to the cells as evidenced by retarded cell growth in the defined medium. Recombinant E. coli strains harboring plasmids which carry the tktA gene showed a tendency to segregate their plasmids almost completely within 24h.

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Production of L-Tryptophan by Auxotrophs Derived from Analogue- resistant Mutants of Escherichia coli (영양요구성 대장균 변이주를 이용한 L-트립토판 생산)

  • Lee, In-Young;Kim, Jae-Hi;Kwak, Moo-Young;Lee, Hosull;Lee, Sun-Bok
    • Microbiology and Biotechnology Letters
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    • v.16 no.5
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    • pp.407-412
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    • 1988
  • In order to increase the tryptophan productivity of E. coli SB1007, a mutant resistant to sulfanilamide was isolated and then a tyrosine auxotroph TY-90 was derived from the sulfanilamide-resistant mutant SA3-39-16. In the test-tube culture a quantitative amount of tryptophan was accumulated in strain TY-90 but in a jar fermentor culture the productivity was lower as compared to the level obtained by the parent strain. From the double auxotrophic mutant SB2756, a revertant resistant to 2, 000$\mu\textrm{g}$/$m\ell$ of $\beta$-thienylalanine, TA 40-10, was selected and then phenylalanine auxotrophs were derived from the revertant strain TA-40-10. One of the phenylalanine auxotrophs, TP-4, accumulated 3.7g/$\ell$ of L-tryptophan after 71-hr cultivation in a jar fermentor experiment.

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Photosensitized Lysis of Egg Lecithin Liposomes by L-Tryptophan and N-Acetylphenylalanyl-L-Tryptophan

  • Cho, Dae-Won;Yoon, Min-Joong
    • Bulletin of the Korean Chemical Society
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    • v.7 no.1
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    • pp.78-81
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    • 1986
  • The photosensitized lysis of egg lecithin lipid membranes (liposomes) have been performed to UV-B light (270-320 nm) by L-tryptophan(L-Trp) and its peptide such as N-acetylphenylalanyl-L-tryptophan(NAPT) incorporated in the liposomes(ca. 0.1% by weight) or in the external buffer (0.1-0.3 mM). Requirement of oxygenation suggests that the lysis of liposomes is caused by the photosensitized oxidation of lipids. There was significant protection against lysis photosensitized by Trp in the external buffer by low concentration of ferricyanide (0.8 mM), but there was no effect on the lytic efficiency by $N_3^-$ which is singlet oxygen($^1O_2$) quencher, indicative of an electron transfer mechanism involved in the photosensitization. The small change of the lytic efficiency with increasing pH from 4 to 9 was interpreted by large target theory and subsequently indicates that superoxide($O_2^-$) may be an active intermediate for the oxidation. The efficiency of photosensitization of Trp was higher than that of NAPT under the same experimental condition. The weak lytic efficiency of liposomes photosensitized by NAPT was enhanced by incorporating NAPT in liposomes, but it was again quenched by ${\beta}$-carotene incorporated in the bilayer of liposomes. These results indicate that a portion of liposome lysis may be due to $^1O_2$ formation from the excited NAPT.

Adsorption Characteristic of L-tryptophan of Affinity Membrane (친화막의 L-tryptophan 흡착특성)

  • Byun, Hong-Sik;Hong, Byung-Pyo
    • Membrane Journal
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    • v.18 no.3
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    • pp.214-218
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    • 2008
  • Protain affinity membranes based on PES-BSA was preapared by the electrospinning method. The process problem caused by the electrospining was solved by using HFB having high solubility and boiling point. It was expecting that the mass production of protein affinity membrane would be possible with broad range of optimized temperature and humidity. BSA in the PES nanofiber was confirmed by the color change from colorless to violet during the biuret test. The buffer solution with DMSO showed that the amount of elution was 5 times higher than the one when the buffer solution without DMSO was used. This is due to the restriction effect of DMSO on the dissociation of L-tryptophan from BSA during the washing step.

Optimization of Indole-3-acetic Acid (IAA) Production by Bacillus megaterium BM5

  • Lee, Jae-Chan;Whang, Kyung-Sook
    • Korean Journal of Soil Science and Fertilizer
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    • v.49 no.5
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    • pp.461-468
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    • 2016
  • One of the important phytohormones produced by plant growth promoting bacteria is the auxin; indole-3-acetic acid (IAA), with L-tryptophan as the precursor. In this study, we focused on the investigation of optimal conditions for the production of IAA by Bacillus megaterium BM5. We investigated culturing conditions, such as incubation temperature, pH of the culture medium and incubation period, with varying media components such as inoculation volume, tryptophan concentration and carbon and nitrogen source. Besides, optimization study intended for high IAA production was carried out with fermentation parameters such as rpm and aeration. The initial yield of $42{\mu}g\;IAA\;ml^{-1}$ after 24 hr increased to $85{\mu}g\;ml^{-1}$ when 5% (v/v) of L-tryptophan was used in the culture broth. The maximum yield of $320{\mu}g\;IAA\;ml^{-1}$ was observed in trypticase soy broth (TSB) supplemented with starch and soybean meal as C and N sources with a C/N ratio of 3:1 (v/v) at $30^{\circ}C$, pH 8.0 for 48 hrs with 1.0 vvm and 250 rpm in 5 L working volume using 10 L scale fermenter. The bacterial auxin extracted from the culture broth was confirmed by thin layer chromatography and high-performance liquid chromatography and effect on plant growth was confirmed by root elongation test.

A Comparison of Inhibitory Effects in Brown and White Rice ( Oryza sativa L. ) against Mutagenicity Induced by Tryptophan Pyrolysates (트립토판 열분해물 유발 변이원성에 대한 현미 및 백미의 억제 효과 비교)

  • Jo, Jeong-Sun;Yu, Jeong-Eun;Jeon, Hyang-Suk
    • Journal of the Korean Dietetic Association
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    • v.3 no.2
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    • pp.105-111
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    • 1997
  • The inhibitory effect of rice(Oryza sartiva L., illpumbyeo) against mutagenicity induced by tryptophan pyrolysates were investigated using Salmonella typhimurium reversion assay. Both methanol extracts of obtained from brown and white rice were found to possess strong activites of inhibiting the mutagenicities of 3-amino-1,4-dimethyl-5H-pyriod[4,3-b]indol(Trp-P-1) and 3-amino-1-methyl-5H-pyrido[4,3-b]indol(Trp-P-2) on Salmonella typhimurium reversion assay. As the concentration of methanol extract increased, inhibitory effect on mutagenicity increased but reached at steady state as inhibition rate of 90% when the concentration was above 10mg/plate. There was no significant difference(p>0.05) in inhibitory effect of methanol extracts between brown and white rice against tryptophan pyrolysates.

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