• Journal of Microbiology and Biotechnology
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• 제14권5호
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• pp.891-900
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• 2004

UDP-N-Acetylglucosamine(GlcNAc):$\beta$1,4-D-mannoside$\beta$-l ,4N-acetylglucosaminyltransferase-III (GnT-III) and UDP-N-GlcNAc:$\alpha$-6-D-mannosid$\beta$-1,6N-acetylglucosaminyltransferase-V(GnT - V) activities were determined in human hepatoma cell lines and metastatic colon cancer cells, and their activities were compared with those of normal liver cells and fetal hepatocytes. GnT-III activities were higher than those of GnT-V in hepatic carcinoma cells. When the two enzyme activities were assayed in highly metastatic colon cancer cells, GnT - V activities were much higher than those of GnT-III. When GlcN, GlcN-biant-PA and UDP-GlcNAc were used as substrates, the enzymes displayed different kinetic properties between hepatic and colon cancer cells, depending on their metastatic potentials. Normal cells of two origins had characteristically very low levels of GnT-III and -V activities, whereas hepatoma and colon cancer cells contained high levels of activities. These data were supported by RT-PCR and Northern blot analyses, showing that the expression of GnT-III and -V mRNAs were increased in proportion to the enzymatic activities. The increased GnT-III, md -V activities were also correlated with increased glycosylation of the cellular glycoproteins in hepatoma and colon cancer cells, as examined by lectin blotting analysis by using wheat germ glutinin (WGA), erythroagglutinating phytohemagglutinin (E-PHA), leukoagglutinating phytohemagglutinin (L-PHA), and concanavalin A (Con A). Treatment with retinoic acid, a differentiation agent, resulted in decreases of both GnT-III and -V activities of HepG2 and HepG3 cells. In colon carcinoma cells, however, treatment with retinoic acid resulted in a reduction of GnT-V activity, but not with GnT-III activity. Although the mechanism underlying the induction of these mzymes is unclear, oligosaccharides in many glycoproteins have been observed of cancer cells.

• Journal of Microbiology and Biotechnology
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• 제31권6호
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• pp.855-866
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• 2021

The effects of various carbon sources on mycelial growth and polysaccharide synthesis of the medicinal fungus Inonotus obliquus in liquid fermentation were investigated. After 12-d fermentation, mycelial biomass, polysaccharide yield, and polysaccharide content were significantly higher in Glc+Lac group (glucose and lactose used as combined carbon source) than in other groups. Crude polysaccharides (CIOPs) and the derivative neutral polysaccharides (NIOPs) were obtained from mycelia fermented using Glc, fructose (Fru), Lac, or Glc+Lac as carbon source. Molecular weights of four NIOPs (termed as NIOPG, NIOPF, NIOPL, and NIOPGL) were respectively 780.90, 1105.00, 25.32, and 10.28 kDa. Monosaccharide composition analyses revealed that NIOPs were composed of Glc, Man, and Gal at different molar ratios. The NIOPs were classified as α-type heteropolysaccharides with 1→2, 1→3, 1→4, 1→6 linkages in differing proportions. In in vitro cell proliferation assays, viability of RAW264.7 macrophages was more strongly enhanced by NIOPL or NIOPGL than by NIOPG or NIOPF, and proliferation of HeLa or S180 tumor cells was more strongly inhibited by NIOPG or NIOPGL than by NIOPF or NIOPL, indicating that immune-enhancing and anti-tumor activities of NIOPs were substantially affected by carbon source. qRT-PCR analysis revealed that expression levels of phosphoglucose isomerase (PGI) and UDP-Glc 4-epimerase (UGE), two key genes involved in polysaccharide synthesis, varied depending on carbon source. Our findings, taken together, clearly demonstrate that carbon source plays an essential role in determining structure and activities of I. obliquus polysaccharides by regulating expression of key genes in polysaccharide biosynthetic pathway.

• Molecules and Cells
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• 제19권2호
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• pp.256-261
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• 2005

In silico analysis of genome of the cyanobacterium Synechocystis sp. PCC 6803 identified two genes, slr0329 and sll0593, that might participate in glucose (Glc) phosphorylation (www.kazusa.or.jp/cyano). In order to determine the functions of these two genes, we generated deletion mutants, and analyzed their phenotypes and enzymatic activities. In the presence of 10 mM Glc, wild-type (WT) and slr0329 defective strain (M1) grew fast with increased respiratory activity and NADPH production, whereas the sll0593 deletion mutant (M2) failed to show any of the Glc responses. WT and M1 were not significantly different in their glucokinase activity, but M2 had 90% less activity. Therefore, we propose that Sll0593 plays a major role in the phosphorylation of glucose in Synechocystis cells.

• 분석과학
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• 제7권1호
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• pp.53-64
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• 1994

아미노산의 광학 이성질체를 분리하는 방법을 확립하고 식품이나 생체에 있는 광학이성질체를 분리 정량하여 영양학이나 생화학적 관계를 연구한다. 필수아미노산 20종을 포함하여 총 38종의 d, l-from 아미노산을 검색 대상으로 삼았다. 단백질은 한국산 콩, 된장, 고추장, 간장, 분유 및 환자의 백내장을 시료로 하여 산 가수분해 후 esterification과 acylation으로 TFA-IPA 유도체를 만들어 chirasil val GLC column을 사용하여 분리하였다. 이성질화가 일어나는 아미노산은 alanine, aspartic acid, glutamic acid, phenyl alanine으로 식물성 시료인 된장에서는 d-alanine이 검출되었으나 동물성 시료인 백내장과 분유에서는 검출되지 않았다. 연령추정 등 생리적으로 이용이 가장 적적한 것은 분리능과 함량을 비교했을 때 aspartic acid였다. d-from 아미노산의 분율은 가정용 된장이 3~6, 시판용 된장이 약 3%, 간장이 2~4%, 콩이 약 1%, 백내장 시료에서 1~2%, 분유에서 1.0~1.5%로 검출되었으며, 숙성 발효시 광학이성질체의 변환이 현저함을 알 수 있다.

• Animal Bioscience
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• 제34권9호
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• pp.1525-1531
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• 2021

Objective: The objective of this study was to evaluate the antimicrobial effects of fermented Maillard reaction products made by milk proteins (FMRPs) on Clostridium perfringens (C. perfringens), and to elucidate antimicrobial modes of FMRPs on the bacteria, using physiological and morphological analyses. Methods: Antimicrobial effects of FMRPs (whey protein plus galactose fermented by Lactobacillus rhamnosus [L. rhamnosus] 4B15 [Gal-4B15] or Lactobacillus gasseri 4M13 [Gal-4M13], and whey protein plus glucose fermented by L. rhamnosus 4B15 [Glc-4B15] or L. gasseri 4M13 [Glc-4M13]) on C. perfringens were tested by examining growth responses of the pathogen. Iron chelation activity analysis, propidium iodide uptake assay, and morphological analysis with field emission scanning electron microscope (FE-SEM) were conducted to elucidate the modes of antimicrobial activities of FMRPs. Results: When C. perfringens were exposed to the FMRPs, C. perfringens cell counts were decreased (p<0.05) by the all tested FMRPs; iron chelation activities by FMRPs, except for Glc-4M13. Propidium iodide uptake assay indicate that bacterial cellular damage increased in all FMRPs-treated C. perfringens, and it was observed by FE-SEM. Conclusion: These results indicate that the FMRPs can destroy C. perfringens by iron chelation and cell membrane damage. Thus, it could be used in dairy products, and controlling intestinal C. perfringens.

• 한국식품과학회지
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• 제22권5호
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• pp.562-568
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• 1990

조리(調理) 육류(肉類)의 WOF와 관련(關聯)된 휘발성(揮發性) 성분(成分)을 분석(分析)하기 위하여 변형(變形)된 Direct GLC 방법(方法)을 이용(利用)하였다. 이 변형(變形)된 방법(方法)은 기존(旣存)의 다른 방법(方法)과는 달리 수분함량(水分含量)이 많은 시료(試料)로부터 휘발성(揮發性) 성분(成分)을 분리(分離)하는데 유용(有用)하였으며, 특히 저분자량(低分子量)의 포화(飽和) $aldehyde\;(C_c-C_{15})$류(類), 불포화(不飽和)$aldehyde(C_5{\sim}C_9)$류(類)의 연구(硏究)에 적절한 것으로 판명(判明)되었다.

• 한국식품위생안전성학회지
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• 제3권2호
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• pp.65-73
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• 1988

종래의 화학적, 생물학적 분석방법의 단점인 검출한도가 낮은점, 시료전처리의 복잡성, 비경제성 및 비 특이성 등을 극복하기 위해 monoclonal AB를 사용한 ELISA법으로 T-2 toxin 에 대해 특이성있는 새로운 분석법을 개발하여 다음과 같은 결과를 얻었다. 1. 종래의 GLC 및 GC-MS 분석법보다 간편하고 신뢰성이 높으며 검출한계가 0.1ppb인 고감도의 분석법을 개발하였다. 2. 본 분석법을 이용하여 Fusarium spp. 균의 T-2 생산유무를 단시간에 다량의 시료를 검색할 수 있었으며, data의 정확도가 GLC와 유사하며 GLC로는 검색할 수 없는 150ppb이하의 미량함유 시료에서도 T-2 toxin을 검색할 수 있었다. 3. 본 실험에 사용한 F. sporotichioides M-1-1 균주의 밀에 대한 최적 배양조건은 $24~27^{\circ}C$ 2주간임을 알았다.

• Journal of Nutrition and Health
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• 제14권1호
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• pp.16-25
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• 1981

한국에서 사육되고 있는 오리의 식용화를 검토하기 위하여 시도하였다. 시장에서 구입한 오리고기를 일반분석하여 수분 62.87%, 조단백질 19.06%, 조지방 17.05%, 회분 1.02%를 나타냈으며 오리고기 단백질을 GLC로 분석하여 거의 모든 필수아미노산이 함유되어 있음을 알 수 있었고 트리프토판이 제한 아이노산 이었다. 한편 오리고기의 지방산을 GLC로 분석하여 oleic acid가 많았고 linoleic acid도 상당량 함유되었음을 알 수 있었고 지방산의 P/S 비는 3.4를 나타냈다. 그리고 오리고기의 콜레스테롤 함량은 70.5mg%를 나타냈고 인지질도 많은 양이 함유되었으며 특히 레시틴이 많았다. 도살 후 2시간까지는 APT-phosphorus의 유리는 높은 온도에서 훨씬 빨리 되어지며, 근원단백질의 ATPase활성은 EDTA, 금속이온의 농도가 증가하면 억제를 받고 농도가 감소되면 활성은 증가되었다. 오리고기를 상이한 조건에서 조리하고 pepsin을 첨가하여 좋은 소화상태를 알 수 있었다. 이는 오리고기가 동물성 단백질이 풍부하며 소화도 잘 되므로 좋은 동물성 단백질원이 된다고 생각되어 진다.

• Journal of Microbiology and Biotechnology
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• 제22권2호
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• pp.190-198
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• 2012

RNA interference (RNAi) is rapidly becoming a valuable tool in biological studies, as it allows the selective and transient knockdown of protein expression. The short-interfering RNAs (siRNAs) transiently silence gene expression. By contrast, the expressed short-hairpin RNAs induce long-term, stable knockdown of their target gene. Trichoplusia ni (T. ni) cells are widely used for mammalian cell-derived glycoprotein expression using the baculovirus system. However, a suitable shRNA expression system has not been developed yet. We investigated the potency of shRNA-mediated gene expression inhibition using human and Drosophila U6 promoters in T. ni cells. Luciferase, EGFP, and ${\beta}$-N-acetylglucosaminidase (GlcNAcase) were employed as targets to investigate knockdown of specific genes in T. ni cells. Introduction of the shRNA expression vector under the control of human U6 or Drosophila U6 promoter into T. ni cells exhibited the reduced level of luciferase, EGFP, and ${\beta}$-N-acetylglucosaminidase compared with that of untransfected cells. The shRNA was expressed and processed to siRNA in our vector-transfected T. ni cells. GlcNAcase mRNA levels were down-regulated in T. ni cells transfected with shRNA vectors-targeted GlcNAcase as compared with the control vector-treated cells. It implied that our shRNA expression vectors using human and Drosophila U6 promoters were applied in T. ni cells for the specific gene knockdown.

• 한국환경농학회지
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• 제16권4호
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• pp.333-340
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• 1997

기체크로마토그래피를 이용하여 토양과 대두시료중 haloxyfop-R 및 haloxyfop-R-methyl의 잔류 분석법을 개발하였다. 토양 및 콩시료를 산성화한 후 acetone으로 추출, 두 화합물을 동시에 추출하였으며 pH조절 분배법을 이용하여 각 화합물별로 분획화하였다. Haloxyfop-R 분획은 $BF_3$/methanol 시약에 의한 methyl화와 추가의 분배과정을 거쳐 기체크로마토그래피에 공시하였으며 haloxyfop-R-methyl 분획은 Florisil 흡착크로마토그래피로 추가 정제하였다. 기체크로마토그래피/전자포획검출기에 의한 haloxyfop-R-methyl의 최소검출량은 0.01ng이었으며 시료추출액중 불순물에 의한 간섭은 관찰되지 않았고 이에 따른 분석법의 검출한계는 토양과 대두시료에서 각각 0.005㎎/㎏ 및 0.01㎎/㎏이었다. 추출 및 분획화과정중 두 화합물간의 교차오염과 haloxyfop-R-methyl의 가수분해는 관찰되지 않아 화합물별 개별정량이 가능하였다. 분석법의 회수율은 haloxyfop-R의 경우 토양과 대두시료에서 각각 88.2${\pm}$3.9% (n=12) 및 88.3${\pm}$4.0% (n=12) 및 85.6${\pm}$5.6% (n=6)이었다. 개발된 분석법의 검출한계, 회수율 및 분석과정의 편이성을 고려할 때 haloxyfop-R과 haloxfop-R-methyl 잔류분석에 실용적으로 활용될 수 있다고 판단되었다.