• 한국식생활문화학회지
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• 제8권3호
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• pp.267-274
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• 1993

무의 염장과정 중 소금의 침투량, 담금액의 pH와 산도 및 조직감의 변화와 이와 관련된 화학적 변화, 효소활성 변화를 알아보고자 하였다. 그 결과는 다음과 같다. 1. 3%와 10% 소금용액에서 생무와 열처리한 무를 담구어 4일간 저장하였을 때, 무 조직 내의 소금농도는 2일까지는 빠르게 증가하였으며 침투속도는 10% 소금용액에서 더욱 컸으며 열처리구에서 더 높았다. 2. 무의 염장과정 중 담금액의 pH는 낮아지고 산도는 증가하여 염장 10일째에 pH는 3.45, 산도는 0.52%를 나타내었다. 3. 힘-거리곡선의 형태는 생무의 경우, 파열점(a)과 최대 압착변형점(c)이 일치하거나 a 이후에 더 큰 c가 나타나서 positive slope을 갖는 곡선이 주종을 이루나, 염장한 무는 a와 c가 일치하면서 특징적인 하나의 peak만을 나타내거나 혹은 a 이후에 최대변형력이 작아지면서 negative slope의 곡선이 주로 나타났다. 또한, 염장이 진행됨에 따라 무의 peak 수는 점차 줄어들었다. 압착시험, 침투관통시험 및 절단시험을 통하여 무의 조직감 변화를 측정한 결과, 무의 압착변형력은 염장과정 중 전반적으로 감소하였다. 크기가 다른 탐침을 이용하여 침투관통시험을 행한 결과, 전반적으로 침투관통력은 감소하였으며 탐침의 직경이 클수록 침투관통력은 증가하는 것으로 나타났다. 또한 절단력은 염장과정 중 증가하였다. 이로부터 염장과정 중 무의 견고성은 감소하였으며 무의 질긴정도(toughness)는 증가하는 것으로 생각된다. 4. AIS로부터 펙틴질을 분획하여 그 함량을 측정한 결과, 염장과정 중 HWSP 함량과 HXSP 함량은 증가하였으며 HClSP 함량은 감소하였다. 5. 염장과정 중 효소 활성의 변화는 발효 4일째까지 증가하다가 감소하였으며 무 염장액에서보다 무 고형분에서의 PG 활성이 크게 나타났다. PE 활성은 염장과정 중 무 고형분과 무 염장액에서 계속 감소하였다. 한편, Cx-cellulase은 염장 초기에는 거의 활성을 나타내지 않았으나 염장 2일째부터 활성이 나타나기 시작하였다.

• Journal of Microbiology and Biotechnology
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• 제27권4호
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• pp.791-807
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• 2017

The type II secretion system (T2SS), which transports selected periplasmic proteins across the outer membrane, has rarely been studied in nonpathogens or in organisms classified as Betaproteobacteria. Therefore, we studied Cupriavidus metallidurans (Cme), a facultative chemilithoautotroph. Gel analysis of extracellular proteins revealed no remarkable differences between the wild type and the T2SS mutants. However, enzyme assays revealed that native extracellular alkaline phosphatase is a T2SS substrate, because activity was 10-fold greater for the wild type than a T2SS mutant. In Cme engineered to produce three Ralstonia solanacearum (Rso) exoenzymes, at least 95% of their total activities were extracellular, but unexpectedly high percentages of these exoenzymes remained extracellular in T2SS mutants cultured in rich broth. These conditions appear to permit an alternative secretion process, because neither cell lysis nor periplasmic leakage was observed when Cme produced a Pectobacterium carotovorum exoenzyme, and wild-type Cme cultured in minimal medium secreted 98% of Rso polygalacturonase, but 92% of this exoenzyme remained intracellular in T2SS mutants. We concluded that Cme has a functional T2SS despite lacking any abundant native T2SS substrates. The efficient secretion of three foreign exoenzymes by Cme is remarkable, but so too is the indication of an alternative secretion process in rich culture conditions. When not transiting the T2SS, we suggest that Rso exoenzymes are probably selectively packaged into outer membrane vesicles. Phylogenetic analysis of T2SS proteins supports the existence of at least three T2SS subfamilies, and we propose that Cme, as a representative of the Betaproteobacteria, could become a new useful model system for studying T2SS substrate specificity.

• 한국식품영양과학회지
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• 제23권3호
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• pp.443-452
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• 1994

This study describes the effects of novel1, 25-dihydroxyvitamin D$_3$ analong[1,25(OH)$_2$-16ene-23yne-26, 27-F6-D$_3$] on proliferation of the human histiocytic lymphoma cell line U937 in vitro. We also examined the expression of c-myc oncogene in U937 cells was apparently inhibited to 62% and 87% of the control level after 4 days in the presence of 10-8M and 10-7 M of this analog, respectively. This compound morpholgically and functionally differentiated U937 cells to nonocyte-macrophage phenotype showing the increase of adherence ability to surface and a decrease of N/C ratio in Giemsa staining . Especially, nonspecific esterase activity which is a marker of cell differentiation to monocyte-macrophage was positive, and production of the positive stained cells increased in a dose dependent fashion . The expression of c-myc oncogene by 1, 25(OH)$_2$D$_3$ analog(10-7 M) was reduced by 60% at the mRNA level as determined by Northern blotting. The effects of this novel analog on cell proliferation and cell differentiation may open op new therapeutic strategies for human disorders such as psoriassis and may provide a tool to understand the mechanism of action of vitamin D$_3$ seco-steroids in malignancy.

• Journal of Acupuncture Research
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• 제23권3호
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• pp.103-115
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• 2006

Alzheimer's disease (AD) is the most prevalent form of neurodegenerative disease associated with aging in the human population. This disease is characterized by the following 4 structural changes : Atrophy of the Cortex, Parasympathetic, and other neural cells, the existence of Neurofibrillary tangles (NFTs), and the accumulation of Senile plaques. NFTs and Senile plaques is known to be the index of this disease. Senile plaques disturbs the neutro transmission and depletes of Acetylcholine. So, Recovery of Acetylcholine is the primal objective for treating Alzheimer's disease. So, Inhibiting the activity of Acetylcholine Esterase (AChE), which causes the hydrolysus of acetylcholine into choline and acetate, can be seen as a key role for treating Alzheimer's disease. Increasing body of evidence has been demonstrated that Bee Venom Acupuncture (BV) could compete with complex protein involving in multiple step of $NF-_{\kappa}B$ activation and exert the anti -inflammatory potential of combined inhibition of the prostanoid and nitric oxide synthesis systems by inhibition of IKK and $NF-_{\kappa}B$. The effect of BV through behavioral tests after memory impairment induced by Scopolamine. We examined the improving effect of BV on the Scopolamine (1 mg/Kg, i.p.)-induced memorial impairment using passive avoidance response and water maze tests in the mice. BV (0.84, $1.67\;{\mu}g/ml$) reversed the Scopolamine-induced memorial impairment in dose dependent manner. This study therefore suggests that BV acupuncture method may be useful for prevention of development or progression of AD.

• Journal of Microbiology and Biotechnology
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• 제10권2호
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• pp.258-263
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• 2000

2-Hydroxy-6-oxo-6phenylhexa-2,4-dienoate (HOPDA) hydrolase catalyzes the hydrolytic cleavage of HOPDA to bemzpate and 2-hydroxypenta-2, 4-dienoate (HPD) during microbial catabolism of biphenyl and polychlorinated biphenyls. A HOPDA hydrolase gene (pcbD) was isolated from the genomic library of Pseudomonas sp. P20 and designated as pCNUO1201; a 7.5-kb XbaI DNA fragment from Pseudomonas sp. P20 was inserted into the pBluescript SK(+) XbaI site. E. coli HB101 harboring pCNU1201 exhibited HOPDA hydrolase activity. The open reading frame (ORF) corresponding to the pcbD gene consisted of 855 base pairs with an ATG initiation codon and a TGA termination codon. The ORF was preceded by a rebosome-binding sequence of 5'-TGGAGC-3' and its G+C content was 55 mol%. The pcbD gene of Pseudomonas sp. P20 was located immedeately downstream of the pcbC gene encoding 2,3- dihydroxybiphenyl 1,2-dioxygenase, and approximately 4-kb upstream of the pcbE gene encoding HPD hydratase. The pcbK gene was able to encode a polypeptide with a molecular weight of 31,732 containing 284 amino acid residues. The deduced amino acid sequence of the HOPDA hydrolase of Pseudomonas sp. P20 exhibited high identity (62%) with those of the HOPDA hydrolases of P. putida KF715, P. pseudoalcaligenes KF707, and Burkholderia cepacia LB400, and also significant homology with those of other hydrolytic enzymes including esterase, transferase, and peptidase.

• Ocean and Polar Research
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• 제31권4호
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• pp.349-357
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• 2009

Ballast water is widely recognized as a serious environmental problem due to the risk of introducing non-indigenous aquatic species. In this study we aimed to investigate measures which can minimize the transfer of aquatic organisms from ballast water. Securing more reliable technologies to determine the viability of aquatic organisms is an important initiative in ballast water management systems. To evaluate the viability of marine phytoplankton, we designed the staining methods of fluorescein diacetate (FDA) and Calcein-AM assay on each target species belonging to different groups, such as bacillariphyceae, dinophyceae, raphidophyceae, chrysophyceae, haptophyceae and chlorophyceae. The FDA method, which is based on measurements of cell esterase activity using a fluorimetric stain, was the best dye for determining live cells of almost all phytoplankton species, except several diatoms tested in this study. On the other hand, although fluorescence of Calcein-AM was very clear for a comparatively longer time, green fluorescence per cell volume was lacking in most of the tested species. According to the Flow CAM method, which is a continuous imaging technique designed to characterize particles, green fluorescence values of stained cells by FDA were significantly higher than those of Calcein-AM treatments and control, implying that the Flow CAM using FDA assay could be adapted as an important tool for distinguishing living cells from dead cells. Our results suggest that the FDA and Calcein-AM methods can be adapted for use on phytoplankton, though species-specific characters are greatly different from one organism to another.

• Journal of Microbiology and Biotechnology
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• 제26권2호
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• pp.248-254
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• 2016

A soil metagenome contains the genomes of all microbes included in a soil sample, including those that cannot be cultured. In this study, soil metagenome libraries were searched for microbial genes exhibiting lipolytic activity and those involved in potential lipid metabolism that could yield valuable products in microorganisms. One of the subclones derived from the original fosmid clone, pELP120, was selected for further analysis. A subclone spanning a 3.3 kb DNA fragment was found to encode for lipase/esterase and contained an additional partial open reading frame encoding a wax ester synthase (WES) motif. Consequently, both pELP120 and the full length of the gene potentially encoding WES were sequenced. To determine if the wes gene encoded a functioning WES protein that produced wax esters, gas chromatography-mass spectroscopy was conducted using ethyl acetate extract from an Escherichia coli strain that expressed the wes gene and was grown with hexadecanol. The ethyl acetate extract from this E. coli strain did indeed produce wax ester compounds of various carbon-chain lengths. DNA sequence analysis of the full-length gene revealed that the gene cluster may be derived from a member of Proteobacteria, whereas the clone does not contain any clear phylogenetic markers. These results suggest that the wes gene discovered in this study encodes a functional protein in E. coli and produces wax esters through a heterologous expression system.

• 한국환경농학회지
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• 제6권2호
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• pp.1-11
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• 1987

수도용(水稻用) 유기인계(有機燐系) 살충제중에서 조제형태(粗劑形態)로 널리 사용되는 Diazinon제와 Dursban제의 담수토양중(湛水土壤中)에서의 경시적(經時的) 분해(分解)정도를 비교(比較)하고, 아울러 살균(殺菌)과 비살균(非殺菌)토양을 비교하므로서 토양미생물(土壤微生物)에 의한 분해효과(分解效果)를 실험하였다. 아울러 두 약제의 분해대사산물을 GC/MS로 확인하였으며 중요한 결과는 아래와 같다. 1. 항온기(恒溫器$(30{\pm}1^{\circ}C)$)내에서 담수상태(湛水狀態)로 처리한 Diazinon제와 Dursban제의 경시적 변화는 비살균토양(非殺菌土壤)에서 보다 살균토양(殺菌土壤)에서 약 3배(培) 정도 지연되었으며, 따라서 미생물에 의한 분해를 확인할 수 있었다. 2. Diazinon제의 상용농도에서의 반감기는 2.2일이었으며, Dursban 제는 10.8일이었다. 상용농도의 3 배량 고농도 처리에서는 두 약제 모두 평균 1일정도 분해가 지연되었다. 3. Diazinon의 분해대사산물(分解代謝産物)로는 가수분해산물인 0, 0-diethyl phosphorothioate 와 이 화합물의 이량체(二量體)인 sulfotep, 그리고 monooxygenase 에 의한 분해대사물인 Diazoxon, 0,0-diethyl-0-[2-(1-hydroxy-1, 1-dimethyl)-6-methyl]-pyrimidinyl phosphorothioate 그리고 2-isopropyl-6-methyl-pyrimidine-4-one 이 확인되었으며, Dursban 제의 대사산물로는 0, 0-diethyl phosphorothioate 만이 확인되어 주로 esterase 에 의한 분해가 주대 대사경로라고 밝혀졌다.

• Current Research on Agriculture and Life Sciences
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• 제4권
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• pp.42-49
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• 1986

Tomato과실(果實)의 성숙단계별(成熟段階別), pectin효소(酵素)(PE, PG)의 활성(活性)의 변화(變化)와 polygalacturonase (PG)의 분리(分離), 정제(精製) 및 정제(精製)된 PG isoenzyme의 효소학적(酵素學的) 성질(性質)에 대하여 조사(調査)하였다. Tomato과실(果實)의 pectin esterase (PE)효소(酵素)는 추숙(追熟)이 진행(進行)됨에 따라 그 활성(活性)이 증가(增加)하였으며, 특히 색깔의 변화가 일어나는 시기(時期)에 최대(最大)의 활성(活性)을 나타내었다. PG는 녹숙과(綠熟果)에서 활성(活性)이 거의 나타나지 않았으나 추숙(追熟)이 진행(進行)되면서 크게 증가(增加)하였다. Ripe tomato에서 추출(抽出)한 조효소액(粗酵素液)을 전기영동(電氣泳動)시킨 결과(結果) 6개의 단백질(蛋白質) band를 나타내었다. 조효소액(粗酵素液)을 황산암모니아에 의한 단백질(蛋白質)의 분획(分劃) DEAE-sephadex A-50 column chromatography한 결과(結果) 2개의 isoenzyme I와 II (PG I, PG II)를 분리(分離)하였으며, isoenzyme I와 II는 각각 61배, 98배 정제(精製)되었다. Isoenzyme I와 II는 전기영동상(電氣泳動上) Rm값은 각각 0.25와 0.31이었다. Isoenzyme I의 최적온도(最適溫度) 및 pH는 각각 $37^{\circ}C$, pH 4.5에서, isoenzyme II는 각각 $40^{\circ}C$, pH 4.7에서 최대활성(最大活性)을 나타내었다. Isozyme I에 대한 pH와 열안정성(熱安定性)을 조사(調査)한 결과(結果), pH 4.3부근에서 안정(安定)하였으며, $78^{\circ}C$, 5분간(分間)에서 약 50%의 효소활성(酵素活性)이 억제되었다. 그리고 isozyme II에 대한 pH와 열안정성(熱安定性)을 조사(調査)한 결과(結果), pH 5.6 부근에서 안정(安定)하였으며 $57^{\circ}C$, 5분간(分間)에서 약 50%의 효소활성(酵素活性)이 억제(抑制)되었다. Isozyme I와 II에 대한 최적(最適) NaCl농도(濃度)는 각각 0.15M, 0.1M이었으며, 최적(最適) $MgCl_2$ 농도(濃度)는 각각 0.04M, 0.03M이었다.

• 고려인삼학회:학술대회논문집
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• 고려인삼학회 2006년도 춘계학술대회
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• pp.3-12
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• 2006

1 The present work was performed to investigate the effects of ginsenoside Rh2 on proliferation, cell cycle-regulation and differentiation of human leukemia HL-60 cells as well as the underlying mechanisms for these effects. 2 Ginsenoside Rh2 potently inhibited the proliferation of HL-60 cells in both a dose- and time-dependent manner with an $IC_{50}$, $20{\mu}M$. 3 DNA flow-cytometry indicated that ginsenoside Rh2 markedly induced a $G_1$ phase arrest of HL-60 cells. 4 Among the $G_1$ phase cell cycle-related proteins, the levels of cyclin-dependent kinase(CDK)4, 6 and cyclin D1, cyclin D2, cyclin D3 were reduced by ginsenoside Rh2, whereas the steadystate levels of CDK2 and cyclin E were unaffected. 5 The protein levels of a CDK inhibitor p16, $p21^{CIP1/WAF1}$ and $p27^{KIP1}$ were markedly increased by ginsenoside Rh2. 6 Ginsenoside Rh2 markedly enhanced the binding of $p21^{CIP1/WAF1}$ and $p27^{KIP1}$ with CDK2 and CDK6, resulting in the reduced activity of both kinases and the hypophosphorylation of Rb protein. 7 We furthermore suggest that ginsenoside Rh2 is a potent inducer of the differentiation of HL-60 cells, based on observations such as a reduction of the nitroblue tetrazolium level, an increase in the esterase activities and phagocytic activity, morphology changes, and the expression of CD11b, CD14, CD64 and CD66b surface antigens. 8 In conclusion, the onset of ginsenoside Rh2-induced the $G_0/G_1$ arrest of HL-60 cells prior to the differentiation is linked to a sharp up-regulation of the $p21^{CIP1/WAF1}$ level and a decrease in the CDK2, CDK4 and CDK6 activities. This is the first report demonstrating that ginsenoside Rh2 potently inhibits the proliferation of human promyelocytic HL-60 cells via the $G_1$ phase cell cycle arrest and differentiation induction.