• 한국동물학회지
• /
• 제30권4호
• /
• pp.432-444
• /
• 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.

• Journal of Microbiology and Biotechnology
• /
• 제11권5호
• /
• pp.798-803
• /
• 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.

• 한국미생물·생명공학회지
• /
• 제16권6호
• /
• pp.450-456
• /
• 1988

대장균 변이주를 이용한 L-트립토판의 최적생산을 위해 목적산물의 생산과 유기산의 생성과의 상호관계를 조사한 결과 비 산생성속도(specific acid production rate)가 증가할수록 L-트립토판의 생산이 감소하는 것으로 나타났다. 따라서 L-트립토판 발효시 산의 생성량을 줄이기 위해 조절식 유가배양법을 도입하였는데 이 배양공정에서는 배양액내의 용존산소농도에 따라 영양배지의 유가속도를 조절하는 방식을 사용하여 세포증식속도를 제한, 세포의 산소요구량이 공급량을 초과하지 않도록 하였다. 이러한 조절식 유가배양법으로 대장균 세포를 배양한 결과 기존의 유가배양식에 비해 비 산생성속도가 현저하게 감소하였으며 L-트립토판의 생산은 5배 정도나 증가되었다.

• Bulletin of the Korean Chemical Society
• /
• 제29권5호
• /
• pp.928-932
• /
• 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.

• Journal of Microbiology and Biotechnology
• /
• 제10권6호
• /
• pp.789-796
• /
• 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.

• 한국미생물·생명공학회지
• /
• 제16권5호
• /
• pp.407-412
• /
• 1988

트립토판 생산성을 향상시키기 위해 대장균 변이 주인 SB1007로부터 sulfanilamide 내성 균주를 유도하여 SA 3-39-16출 분리하고 이 균주로부터 tyrosine 영양요구주인 TY-90을 개발하였다. 시험관배양시 TY-90균주는 모균주보다 트립토판 생산량이 우수하였으나 발효조 배양시에는 모균주에 비해 낮은 양의 트립토판이 축적되었다. 발효조에서의 생산성 증대를 인해 또 다른 대장균 변이주인 SB2756으로부터 새로운 변이주의 개발을 모색하였다. 먼저 이중 영양요구성 균주인 SB2756의 revertant를 분리하여 그중 2,000mg/$\ell$ 농도의 $\beta$-thenylalanine에 내성을 갖는 TA-40-10을 분리하였고 이 균주로부터 phenylalanine 영향요구주인 TP-4를 개발하였다. 이 TP-4균주는 발효조에서 71시간 배양시 3.7g/$\ell$의 L-tryptophan을 생산하였다.

• Bulletin of the Korean Chemical Society
• /
• 제7권1호
• /
• pp.78-81
• /
• 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.

• 멤브레인
• /
• 제18권3호
• /
• pp.214-218
• /
• 2008

전기방사를 이용하여 polyethersulfone (PES)-bovin albumin serum (BSA) 단백질 친화막을 제조하였으며, 이때 전기방사에 의한 공정상의 문제점을 용해도가 높고, 끓는점이 높은 2,2,3,4,4,4-Hexafluoro-1-butanol (HFB)를 공용매로 사용함으로써 해결하였다. 또한 공용매는 최적의 온도, 습도 범위를 확대시켜줌으로써 친화막의 대량 생산이 용이하리라 생각한다. 제조된 친화막은 biuret test를 통하여 친화막의 색깔이 무색에서 보라색으로 변함으로써 PES 섬유 내 BSA가 존재함을 확인할 수 있었다. 완충용액의 조성 성분 중 dimethyl sulfoxide (DMSO)는 세척과정에서 BSA와 L-tryptophan사이의 해리를 제어함으로써 용출과정에서 비교실험보다 약 5배의 높은 용출량을 보여주었다.

• 한국토양비료학회지
• /
• 제49권5호
• /
• pp.461-468
• /
• 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.

• 대한영양사협회학술지
• /
• 제3권2호
• /
• pp.105-111
• /
• 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.