• 한국수정란이식학회지
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• 제30권4호
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• pp.315-317
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• 2015

In vitro culture of murine embryos is an important step for in vitro production systems including in vitro fertilization and generations of genetically engineered mice. M16 is widely used commercialized culture media for the murine embryos. Compared to other media such as potassium simplex optimization medium, commercial M16 (Sigma) media lacks of amino acid, glutamine and antibiotics. In the present study, we optimized M16 based embryo culture system using commercialized antibiotics-glutamine or amino acids supplements. In vivo derived murine zygote were M16 media were supplemented with commercial Penicillin-Streptomycin-Glutamine solution (PSG; Gibco) or MEM Non-Essential Amino Acids solution (NEAA; Gibco) as experimental design. Addition of PSG did not improved cleavage and blastocyst rates. On the other hand, cleavage rate is not different between control and NEAA treated group, however, blastocyst formation is significantly (P<0.05) improved in NEAA treated group. Developmental competence between PSG and NEAA treated groups were also compared. Between two groups, cleavage rate was similar. However, blastocyst formation rate is significantly improved in NEAA treated group. Taken together, beneficial effect of NEAA on murine embryos development was confirmed. Effect of antibiotics and glutamine addition to M16 media is still not clear in the study.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2002년도 생물공학의 동향 (X)
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• pp.309-312
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• 2002

GS system을 이용하여 재조합 EPO를 생산하는 새로운 CHO-K1 세포주를 확립하였으며, 이 세포주를 이용하여 Zn 이온과 Mn 이온이 EPO의 생산에 미치는 영향에 관해서 연구하였다. 형질 전환에 있어서 DNA 3 ${\mu}g$ 올 사용하고, 세포군 선별을 위한 MSX의 농도는 100 11M 을 사용한 경우에만 세포군이 발견되었다. 200 ${\mu}M$의 MSX를 처리한 경우에서는 세포군이 생성되지 않았고, 이는 고농도의 MSX에 의해 세포 생장이 저해되었기 때문이다. 형질 전환된 CHO-K1 세포 배양에 Zn 이온을 첨가한 결과 세포의 생장은 크게 저해되지 않은 반연에, EPO의 생산은 대조구에 비해 40% 이상 증가함을 알 수 있었다. 이러한 결과는 Mn 이온을 처리한 실험에서도 관찰할 수 있었다.

• Applied Biological Chemistry
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• 제25권3호
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• pp.173-176
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• 1982

느릅나무근(根) 점액(粘液)은 단백질(蛋白質)을 함유(含有)하며 그 함량(含量)은 황촉규근(黃蜀葵根) 점액(粘液)의 단백질(蛋白質) 함량(含量)보다 크다. 점액(粘液)에서 alanine, aspartic acid, glutamine, glutamic acid, hydroxyproline, isoleucine, leucine, proline, phenylalanine 및 tyrsine 등(等) 아미노산(酸_이 검출(檢出)되고 또 이 점액(粘液)에 무수(無水) ethanol을 가(加)하여 얻은 점질물(粘質物)의 가수분해(加水分解) 생성물(生成物)에서는 alanine, aspargine, aspartic acid, glutamine, glutamic acid, hydroxyproline, isoleucine, leucine, methionine, proline, phenylalanine, serine, tyrosine 그리고 valine등(等) 여러 종류(種類)의 아미노산(酸)이 검출(檢出)되었다.

• 한국미생물·생명공학회지
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• 제19권3호
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• pp.259-264
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• 1991

K.pneumoniae의 nif 유전자 발현에 미치는 glnA 산물 (GS)의 영향을 살펴보기 앞서 K.pneumoniae의 GS를 분리.정제하여 그 성질을 조사하였다. K.pneumoniae의 GS는 분자량 약 600,000의 12개 동일 subunit으로 구성되어져 이었으며 최적 반응 pH 및 온도는 각기 pH 7.0, $37^{\circ}C$ 였고 pH 5-8사이에서 활성의 변화가 크지 않았으며 $57^{\circ}C$에서 10분간 열처리 할 때 50의 활성 감소가 나타났다. 또 암모니아 농도의 변화에 따라 adenylylation-deadenylylation 기작에 의한 활성조절기작이 E.coli GS와 동일 했으며 anti-E.coli GS 항체를 이용한 immunodiffusion에서도 K.pneumoniae GS가 E.coli GS와 혈청적으로 매우 유사함이 드러났다.

• 식물조직배양학회지
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• 제26권2호
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• pp.99-102
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• 1999

Glucose - induced arginine transport system was induced by a exogenous application of NH$_4$^+ ion. The uptake rate of arginine (Arg) depended on the external NH$_4$^+ ion concentration. The uptake rate was inhibited by the presence of NH$_4$^+ ion within 1 min, whereas it increased maximally after 30 min. Glucose and NH$_4$^+ ion induced the same arginine transport system. Km value of Arg transport systems was 2 $\mu$M, and V_(max) was 60 $\mu$mol^(-1) . h . g fresh weight^(-1) for NH$_4$^+ ion and 174 $\mu$mol^(-1) . h . g fresh weight^(-1) for glucose induced transport system. But, the transport system of Glu for glucose and NH$_4$^(-1) ion induced had different Km values. Km value of Glu was 285 $\mu$M for glucose - and 58 $\mu$M for NH$_4$^+ ion induced transport system. Thus, NH$_4$^+ ions play a important role as inducer for the glutamine transport system. NH$_4$^+ ion induced glutamine system was inhibited over 90% by cycloheximide. We concluded that a new carrier protein for glutamine was induced by NH$_4$^+ ion.

• 대한화학회지
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• 제34권6호
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• pp.622-628
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• 1990

장미 구성요소 중 꽃잎 부분을 $NH_3$ 감응센서에 고정시켜서 글루타민 조직센서를 제조하여 pH, 완충용액, 조직 양 및 여러 가지 방해물질의 영향과 전극의 수명 등에 관하여 조사하였다. 그 결과 37$^{\circ}C$에서 pH 7.8, 0.2M 인산완충용액을 사용하였을 때, $8.0 {\times} 10^{-4}$ ∼$5.0 {\times} 10^{-2}$ M 글루타민 직선범위와 52 mV/decade의 감응기울기를 나타내었다. 이 때 사용한 조직 양은 50 mg이었다. 본 센서는 선택성이 탁월한 것으로 나타났다. 이 센서를 장미의 다른 구성요소와 비교하였다. 실제 글루타민의 정량에 유용하게 사용될 것으로 나타났다.

• 미생물학회지
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• 제24권2호
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• pp.113-118
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• 1986

Optimum temperature and pH of glutamine synthetase activity (E.C. 3.6.1.2.) of R. sphaeroides D-230 was $35^{\circ}C$ and 6.8, respectively. The adenylated state of GS in R. sphaeroides D-230 was stabilized by addition of 0.2mg/ml of cethyltrimethylammoniumbromide. Valine, histidine, proline, isoleucine, and lysine were good nitrogen source for the growth of R. sphaeroides D-230. The growth of R. sphaeroides D-230 in $N_2,\;NaNO_3\;or\;NH_4Cl$ as sole nitrogen source was lower than in any otherculture conditions. GS activity was inhibited, more or less, by various amino acid. THe relative inhibition rate of the enzyme by added 7mM arginine, $NH_4Cl,\;N_2,\;and\;NaNO_3$ was 63.8%, 26.79%, 6.24%, and 10.64%, drespectively. THe hydrogen evolution of R. sphaeroides D-230 grown in N-limited media was inhibited by 0.1mM MSX, irreversible GS inhibitor. GS activity was completely inhibited by 1.0mM MSX but ammonia released maximally at the same concentration of MSX. Ammonia release by added MSX was increased up to 1.0mM MS, but decreased above 1.0mM MSX. It is probably due to inhibition of nitrogenase actixity by MSX. Nitrogenase activity was not inhibited at low concentration of MSX. These results suggests that the inhibition of nitrogenase activity by ammonia is mediated by products of ammonia assimilation rather than by ammonia itself.

• Journal of Nutrition and Health
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• 제32권2호
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• pp.158-165
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• 1999

Effects of oral taurine supplementation (6g/day) on plasma concentration and urinary of free amino acids were evaluated in healthy female adults. Among twenty five female volunteers(23.6$\pm$0.3 years old) participated in the taurine supplementation program, twenty four subjects successfully completed the two supplementation program. Plasma and urinary levels of free amino acids were determined by using an automated amino acid analyzer based on ion-exchange chromatography. Two weeks of taurine supplementation resulted in a 65% increase in plasma taurine concentration (p<0.001), Changes in fasting plasma amino acid concentrations followed by taurine supplementation were not spectacular, and were all within the normal range for human aldults. Taurine supplementation significantly elevated urinary methionine, asparagine, hydorxyproline and phosphoserine excretions(31~280%), and significantly decreased the urinary excretions of isoleucine, glutamate and serine compared to the values prior to taurine supplementation. For almost every individual amino acids, 24 hr urinary excretion level was significantly correlated to the urinary excretion value expressed as nmol/mg creatinine(p<0.001). A significant negative correlation found between plasma glutamine concentration and urinary glutamine excretion level suggests that the decrease in plasma glutamine concentration might be associated with the enhanced glutamine excretion in urine followed by taurine supplementation.

• Asian Pacific Journal of Cancer Prevention
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• 제16권11호
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• pp.4769-4775
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• 2015

Background: Hepatocellular carcinoma (HCC) is the fifth most prevalent cancer and thirdly leading cause of cancer-related death worldwide. The estimated risk of hepatocellular carcinoma is 15 to 20 times as high among persons infected with HCV as it is among those who are not infected, with most of the excess risk limited to those with advanced hepatic fibrosis or cirrhosis. Glypican3 (GPC3) plays a key role in relation to signaling with growth factors, regulating the proliferative activity of cancer cells. Glutamine synthetase (GS) catalyzes the synthesis of glutamine from glutamate and ammonia in the mammalian liver. GS was suggested as a specific marker for tracing cell lineage relationships during hepatocarcinogenesis. In normal liver, GS expression is seen in pericentral hepatocytes, but not by midzonal or periportal hepatocytes. In HCC, strong and diffuse GS expression in seen in tumor cells. Results: Glypican3 immunopositvity was highly specific and sensitive indicator for hepatocellular carcinoma as well as glutamine synthetase which was found to be a sensitive and specific indicator for development of hepatocellular carcinoma when compared to cirrhosis, liver cell dyspalsia and metastatic carcinomas. Statistical analysis revealed a significant association between GPC3 and GS with tumor size (P=0.003, p=0.006, respectively). Diffuse staining significantly associated with large tumor size while, focal and mixed staining was detected more with small tumor size. Studying the relation with tumor grade also revealed significant association between diffuse GPC3 and GS staining with high tumor grade. Diffuse staining was detected in 91.7% and 100% respectively of poorly differentiated specimens and only in 33.3% and 22.2% of well differentiated specimens. Conclusions: While using GPC3 and GS to screen for premalignant hepatic lesions remains controversial, our data suggest that GPC3 and GS may be a reliable diagnostic immunomarkers to distinguish HCC from benign hepatocellular lesions. However, negative immunostaining should not exclude the diagnosis of HCC.

• 한국미생물·생명공학회지
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• 제18권4호
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• pp.401-405
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• 1990

5 혈청을 포함한 DMEM 배지에 갑상세포주 579를 연속배양하여 약 $5.7 \times 10^{-8}$g/h /cell에 해당하는 pro-UK의 비생산속도를 얻을 수 있었다. 또한 배지의 이동속도가 증가할 수록 glutamine의 소비속도가 증가하는 반면 ammonia 생산속도는 일정하게 유지되는, glutamine의 완전동화에의한 물질생산증가 현상을 나타냈다. 5mM의 glucose와 2mM의 glutamine, 포화용존 공기의 10에 해당하는 용존산소 및 pH 6.2의 maintenance 생육 조건하에서 약 15일간 유지시켜, $12\times 10^{-8}$g of pro-UK/h/cell의 최대 비생산속도와 0.226mg/g of glucose의 생산수율을 얻었으며, 이는 10ml/min의 배지 이동속도를 유지하는 연속배양 조건에서 매일 0.223mg의 pro-KU를 생산할 수 있을을 의미한다.