• Journal of Applied Biological Chemistry
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• 제56권2호
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• pp.89-93
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• 2013

Gamma-aminobutyric acid (GABA) aminotransferase (gabT) and succinic semialdehyde dehydrogenase (gabD) genes from Pseudomonas fluorescens KCCM 12537 were cloned into a single pETDuet-1 vector and co-expressed in Escherichia coli BL21(DE3) simultaneously. The mixture of both enzymes, called GABase, is the key enzyme for the enzymatic analysis of GABA. The molecular mass of the GABA aminotransferase and succinic semialdehyde dehydrogenase were determined to be 52.8 and 46.7 kDa following computations performed with the pI/Mw program, respectively. The GABase activity between pH 6.0 and 9.0 for 24 h at $4^{\circ}C$ remained over 75%, but under pH 6.0 decreased rapidly. The GABase activity between 25 and $35^{\circ}C$ by the treatment at pH 8.6 for 30 min remained over 80%, but over $35^{\circ}C$ decreased rapidly. When the activity against GABA was defined as 100%, the purified GABase activity against 5-aminovaleric acid having a similar structure to GABA showed 47.7% and GABase activity against ${\beta}$-alanine, ${\varepsilon}$-amino-n-caproic acid, $_L$-ornithine, $_L$-lysine, and $_L$-aspartic acid showed between 0.3 to 2.3%. The GABA content was analyzed with this co-expressed GABase, compared with the other GABase which was available commercially. As a result, the content of GABA extracted from brown rice, dark brown rice, and black rice were $26.4{\pm}3.5$, $40.5{\pm}4.7$ and $94.7{\pm}9.3{\mu}g/g$, which were similar data of other GABase in the error ranges.

• Journal of Plant Biology
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• 제31권2호
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• pp.91-100
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• 1988

The effect of Ca2+ and polyamines on the activity $\beta$-glucan synthetase II(GSII) related to cell wall synthesis was studied in carrot suspension cultured cells. The activity of GS II is four times higher than that of $\beta$-glucan synthetase I in carrot suspension cultured cells and in vitro expreiment, the activity of GSII was increased in response to increase in concentration of Ca2+ and polyamines. When carrot suspension cultured cells were incubated together with Ca2+ and polyamines, the GSII activity was high at 0.1mM of Ca2+ and 1mM of putrescine. Also, polycationic poly-L-lysine and poly-L-ornithine increased about 50% the GSII activity than that of the control, respectively. These results may imply that Ca2+ and polyamines were related to the enzyme activity as a polycationic nature. In addition, verapamil as the calcium channel blocker and flunarizine as an antagonist of calcium mechanism in cytoplasm decreased GSII activity ramarkably, Ca2+ and calmodulin stimulated GSII activity as Ca2+ of free ion rather than Ca2+ calmodulin complex. The effect of 2,4-D on the GSII activity in culture medium is shown to be low at 0.1mg per liter and GSII activity increased about 30% more than that of the 0.1mg/l at the range of 0.3-1.0mg per litere. Cummulative results suggest that Ca2+ and polyfamines stimulate the cell wall synthesis by means of the enhancement of GSII activity responsible for synthesizing the cell wall components.

• 미생물학회지
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• 제30권6호
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• pp.483-487
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• 1992

Among a few number of UV-resistant isolated form various environmental sources (10), we made a comparative physio-chemoanalytical study on one of spherical bacteria isolated from air dust, presumably Deinococcus sp. (CM strain 29) with an UV resistant bacterium, Deinococcus radiophilus ATCC 27603 as the reference strain. Our isolate of UV resistant coccus, Deinococcus sp. CM 29 and D. radiophilus ATCC 27603 showed more than 75% matching coefficient in metabolic activity of various substrates. The most predominant cellular fatty acid of both strains was palmitoleic acid (C 16 :1, cis 9), but the detail fatty acid profiles were slightly dissimilar to each other. Cell-bound arange pigment seemed to be an identical chemicals on spectrophotometric analysis. L-ornithine was detected as cell-wall amino acid in both strains. Galactose was detected as cell-wall sugar in D. radiophilus ATCC 27603, whereas glucose in Deinococcus sp. CM 29. G-C molar ratio of both strains was comparable, 63-65%.

• Journal of Radiation Protection and Research
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• 제10권2호
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• pp.122-129
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• 1985

S-2, W-diaminopentyl isothiuronium bromide 및 그 thiophosphate 유도체는 phthalimide 또는 l-ornithine 산(酸)으로부터 출발(出發)하여 합성(合成)되었다. 합성(合成)한 isothiuronium bromide 및 S-2-(W-aminoalkylamino)ethyl isothiuronium bromide의 약독성(藥毒性)을 체중(體重) 25-35g, $4{\sim}8$주생(週生) ICR 생쥐를 사용(使用)하여 검사(檢査)하였다. 그 결과(結果) 전자(前者)의 화합물(化合物)이 후자(後者)의 것들보다 약독성(藥毒性)이 약(弱)하다는 것이 판명(判明)되었다. 이와같은 약독성(藥毒性)의 차이(差異)는 화합물(化合物)들의 화학구조(化學構造)의 차(差)에 기인(起因)하는 것으로 보였으며 IR 분광분석법(分光分析法)으로 그 내용(內容)이 검토(檢討)되었다. 이들 화합물(化合物)의 방사선장해방호효과에 관(關)하여 현재(現在)까지의 관련(關聯)된 연구(硏究)문헌에 대한 고찰을 하였으며 diaminopentyl 유도체들은 약독성(藥毒性)이 비교적 낮고 좋은 방사선장해방호화합물(放射線障害防護化合物)인 것으로 사려된다.

• 한국식품영양과학회지
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• 제41권7호
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• pp.927-935
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• 2012

남아도는 오이의 활용도 및 그 부가가치를 높이기 위하여 오이 식초를 제조한 후 이화학적 성분을 분석하고, 항산화 및 숙취해소 효능과 같은 기능성을 조사하였다. 초기당도를 $15^{\circ}Brix$가 되도록 조절하여 $25^{\circ}C$에서 알코올 발효하였을 때, 발효 6일째에 7.8%의 최대 알코올 함량을 얻었으며, 이를 다시 $30^{\circ}C$에서 12일간 초산발효 하여 산도 5.8%의 식초를 얻었다. 오이 식초의 주요 유리당은 glucose 및 fructose로 그 함량은 각각 3,067.26 및 395.73 mg%였고, 주요 유기산은 acetic acid 및 succinic acid로 그 함량은 각각 4410.5 및 841.11 mg%이었다. 오이 식초의 유리아미노산 총 함량은 181.45 ${\mu}g/mL$이었고, 이 중 citrulline, valine, aspartic acid, asparagine 및 ornithine이 주된 아미노산이었으며, 무기성분은 K, Ca, Mg와 같은 알칼리성 원소를 다량 함유하고 있는 것으로 나타났다. 또한 DPPH, $ABTS^+$ 라디칼 소거능, 환원력 및 ${\beta}$-carotene bleaching과 같은 실험방법을 통해 오이 식초의 항산화 활성을 측정한 결과 높은 활성을 나타내었으며, 오이 식초의 주요 항산화 활성성분인 폴리페놀의 함량은 40.14 mg/100 mL이었다. 한편, 오이 식초는 급성으로 알코올을 투여한 흰쥐의 간조직 중 알데히드 탈수소효소 활성을 높임으로써 혈장 중 아세트알데히드 농도를 효과적으로 낮추었다. 따라서 본 연구결과를 통해 오이를 주원료로 하여 제조한 오이 식초는 상당한 항산화 및 숙취해소 효과가 있는 것으로 판단되며, 이를 기능성식품 소재로 활용이 가능하리라 생각된다.

• Biomaterials and Biomechanics in Bioengineering
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• 제1권1호
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• pp.27-39
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• 2014

We have previously described a new multilayer alginate microcapsule system, and the goals of the present study were to assess the in vitro function of islets encapsulated in its inner layer, and the angiogenic ability of FGF-1 delivered from the external layer in an omentum pouch. Following isolation and culture, islets were encapsulated in the inner core of microspheres ($500-600{\mu}m$ in diameter) with a semi-permeable poly-L-ornithine (PLO) membrane separating two alginate layers, and both unencapsulated and encapsulated islet function was assessed by a dynamic glucose perifusion. For angiogenesis experiments, one group of microcapsules without FGF-1 (control) and another (test) containing FGF-1 with heparin encapsulated in the external layer were made. One hundred microcapsules of each group were transplanted in Lewis rats (n = 5/group) and were retrieved after 14 days for assessment of angiogenesis. Glucose perifusion of unencapsulated and encapsulated islets resulted in similar stimulation indices. The release of FGF-1 resulted in increased vascular density compared to controls. In conclusion, islets encapsulated in the core of multilayer alginate microcapsules maintain functionality and the microcapsule's external layer is effective in delivery of FGF-1 to enhance graft neovascularization in a retrievable omentum pouch.

• Mycobiology
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• 제42권4호
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• pp.361-367
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• 2014

Makgeolli, also known as Takju, is a non-filtered traditional Korean alcoholic beverage that contains various floating matter, including yeast cells, which contributes to its high physiological functionality. In the present study, we assessed the levels of ${\beta}$-glucan and glutathione in various yeast strains isolated from traditional Korean Nuruk and selected a ${\beta}$-glucan- and glutathione-rich yeast strain to add value to Makgeolli by enhancing its physiological functionality through increased levels of these compounds. Yeast ${\beta}$-glucan levels ranged from 6.26% to 32.69% (dry basis) and were strongly species-dependent. Dried Saccharomyces cerevisiae isolated from Nuruk contained $25.53{\mu}g/mg$ glutathione, $0.70{\mu}g/mg$ oxidized glutathione, and $11.69{\mu}g/g$ and $47.85{\mu}g/g$ spermidine and L-ornithine monohydrochloride, respectively. To produce functional Makgeolli, a ${\beta}$-glucan- and glutathione-rich yeast strain was selected in a screening analysis. Makgeolli fermented with the selected yeast strain contained higher ${\beta}$-glucan and glutathione levels than commercial Makgeolli. Using the selected yeast strain to produce Makgeolli with high ${\beta}$-glucan and glutathione content may enable the production of functional Makgeolli.

• 대한유전성대사질환학회지
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• 제2권1호
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• pp.77-79
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• 2002

The urea cycle, consisting of a series of six enzymatic reactions, plays key roles to prevent the accumulation of toxic nitrogenous compound and synthesize arginine de novo. Five well characterized diseases have been described, resulting from an enzymatic defect in the biosynthesis of one of the normally expressed enzyme. This presentation will focus on two representative diseases; ornithine transcarbamylase(OTC) deficiency and citrullinemia(argininosuccinate synthetase deficiency). OTC deficiency is one of the most common inborn error of urea cycle, which is inherited in X-linked manner. We identified 17 different mutations in 20 unrelated Korean patients with OTC deficiency; L9X, R26P, R26X, T44I, R92X, G100R, R141Q, G195R, M205T, H214Y, D249G, R277W, F281S, 853 del C, R320X, V323M and 10 bp del at nt. 796-805. These mutations occur at well conserved nucleotide sequences across species or CpG hot spot. The L9X and R26X lead to the disruption of leader sequences, required for directing mitochondrial localization of the OTC precursor. Their phenotypes are severe, and neonatal onset. The G100R, R277W and V323M mutations were uniquely identified in patients with late onset OTC deficiency. The other genotypes are associated with neonatal onset. Out of 20 patients with OTC deficiency, only 6 patients are alive; two were liver transplanted, and normal in growth and development at 2, 4 years after transplantation respectively. Citrullinemia is an autosomal recessive disease, caused by the mutations in the argininosuccinate synthetase(ASS) gene. We identified in 3 major mutations in 11 unrelated Korean patients with citrullinemia; G324S, $IVS6^{-2}$ A to G, and 67 bp ins at nt 1125-1126. Among these, the 67 base pair insertion mutation is novel. The allele frequency of each mutation is; G324S(45%), IVS6-2 A to G(32%), and 67 base pair insertion(14%). All patients are diagnosed at neonatal or infantile age. Interestingly, two patients presented with stroke like episode. Out of 11 patients, 5 patients died. Among 6 patients alive, one patient was successfully liver transplanted.

• 한국식품영양과학회지
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• 제41권1호
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• pp.79-86
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• 2012

본 연구에서는 2단계 발효를 통해 보당 없이 야콘 식초를 제조한 후 이에 대한 이화학적 성분 분석 및 항당뇨 효과를 조사하였다. 1단계 알코올 발효에서는 $28^{\circ}C$에서 6일간 발효하여 알코올 함량 5.2%의 야콘 와인을 얻을 수 있었으며, 2단계 초산발효에서는 $30^{\circ}C$, 200 rpm으로 6일간 발효하여 산도 4.75%의 야콘 식초를 생산할 수 있었다. 야콘 식초의 주요 유리당은 glucose와 fructose로 나타났으며, 유기산은 acetic acid가 가장 높았으며 이어서 succinic acid가 높게 나타났다. 야콘 식초의 유리아미노산 총 함량은 62.88 mg% 로 proline, ${\gamma}$-amino-n-butyric acid, ornithine이 주된 아미노산으로 나타났다. 무기성분은 Ca, K, Mg와 같은 알칼리성 원소를 다량 함유하고 있는 것으로 나타났다. 제2형 당뇨병 마우스에게 야콘 식초를 4주간 경구투여 하여 혈당 강화 효과를 평가한 결과 공복 시 혈당은 5% 야콘 식초군에서 대조군에 비하여 유의적으로 낮았으며(p<0.05) 10% 야콘 착즙액과 10% 야콘 식초군은 낮은 경향이었다. 내당능 역시 5% 야콘 식초군이 대조군에 비하여 유의적으로 개선하였다(p<0.05). 혈장의 인슐린과 C-peptide 농도 및 췌장의 인슐린 농도는 실험군간 차이가 없었으나 췌장의 C-peptide 농도는 대조군에 비하여 야콘 식초에 농도 의존적으로 높은 것으로 나타났다.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2002년도 국제심포지움
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• pp.88.2-98
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• 2002

A wide arry of naturally occurring substances particularly those present in dietary and medicinal plants, have been reported to possess substantial cancer chemopreventive properties. Certain phytochemicals retain strong antioxidative and anti-inflammatory properties which appear to contribute to their chemopreventive or chemoprotective activities. Inducible cyclooxygenase(COX-2) and nitric oxide synthase (iNOS) are important enzymes that mediate inflammatory processes. There is some evidence that expression of both COX-2 and iNOS is co-regulated by the eukaryotic transcription factor NF-$textsc{k}$B. Increased expression of COX-2 and/or iNOS has been associated with pathophysiology of certain types of human cancers as well as inflammatory diseases. Since inflammation is closely linked to tumor promotion, substances with potent anti-inflammatory activies are anticipated to exert chemopreventive effects on carcinogenesis, particularly in the promotion stage. An example is curcumin, a yellow pigment of turmeric (Curcuma longa L., Zingiberaceae), that strongly occurring diaryl heptanoids structurally related to curcumin have substantial anti-tumor promotional activities in two-stage mouse skin carcinogenesis. Thus, yakuchinone A [1-(4'-hydroxy-3'-methoxyphenyl)-7-phenyl-3heptanone] and yakuchinone B [1-(4'-hydroxy-3'methoxyphenyl)-7-phenylhept-1-en-3-one] present in Alpinia oxyphylla Miquel (Zingiberacease) attenuate phorbol ester-induced inflammation and papilloma formation in female ICR mice. These diarylheptanoids also suppressed phorbol ester-induced activation of epdermal ornithine decarboxylase and its mRNA expression when applied onto shaven backs of mice. Yakuchinone A and B as well as curcumin inhibited phorbol ester-induced expression of COX-2 and iNOS and their mRNA in mouse skin via inactivation of NF-$textsc{k}$B. Capsaicin, a major pungent ingredient of red pepper also attenuated phorbol ester-induced NF-$textsc{k}$B activation. Similar suppression of COX-2 and iNOS and down-regulation of NF-$textsc{k}$B activation for its DNA binding were observed with the ginsenosied Rg3 and the ethanol extract of Artemisia asiatica. We have also found that certain anti-inflammatory phytochemicals exert inhibitory effects on phorbol ester-induced COX-2 expression and NF-$textsc{k}$B activation in immortalized human breast epithelial (MCF-10A) cells in culture. One of the plausible mechanisms undelying inhibition by aforementioned phytochemicals of phorbol ester-induced NF-$textsc{k}$B activation involves interference with degragation of the inhibitory unit, I$textsc{k}$Ba, which blocks subsequent nuclear translocation of the functionally active p65 subunit of NF-$textsc{k}$B. the activation of epidermal NF-$textsc{k}$B by phorbol ester and subsequent induction of COX-2 hence appear to play an important role in intracellular signaling pathwasy leading to tumor promotion and targeted inhibition of NF-$textsc{k}$B may provide a new promising cancer chemopreventive strategy.