• 한국식품과학회지
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• 제25권3호
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• pp.197-203
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• 1993

식물성 식품재료를 대상으로 보체 용혈분석법$(TCH_{50})$ 을 이용하여 보체 활성화능(항보체 활성)에 대한 검색을 실시하였다. 총 38종의 식용 식물추출물 중 5종의 시료에서 대조구에 비해 60% 이상의 $TCH_{50}$ 감소를 일으키는 비교적 높은 활성을 나타냈으며 그 활성의 순서는 $1000{\mu}g/ml$ 시료 농도에서 생강>토란대>냉이>은행잎>달래이었다. 한편 가장 높은 활성을 보였던 생강에서 조제된 ZR-1의 경우 pronase 소화 후에는 활성의 변화가 없는 반면에 과요오드산 산화에 의해서는 급격한 활성의 감소를 나타냄으로써 ZR-1의 단백질 부위가 아닌 다당 부위가 활성에 기여함을 알 수 있었다. 또한 ZR-1의 항보체 활성은 $Ca^{2+}$ 이온 부재시 부분적인 감소현상을 보였으며, ZR-1을 정상인의 혈청과 반응 후 anti-humanC3를 이용하여 2차원 면역 전기영동을 행한 결과 C3의 분해산물을 관찰할 수 있었다. 또한 이들 획분은 $ACH_{50}$의 저해를 일으켰다. 동 결과로부터 ZR-1의 보체 활성화 양식은 classical pathway 뿐만 아니라 alternative pathway도 경유함을 알 수 있었다.

• 한국미생물·생명공학회지
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• 제33권4호
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• pp.288-294
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• 2005

다양한 생리활성을 갖는 flavonoid 화합물이 감귤에 비활성 형태의 배당체 화합물로 다량으로 존재하고 있으므로 비활성 형태의 flavonoid 물질을 활성 형태의 flavonoid 무배당체 물질로 변환 시켜 활성 형태의 flavonoid 화합물을 증가하고자 하였다. 무배당체 화합물로 전환하기 위하여 시판되고 있는 당 분해효소 중 amylase가 주성분인 AMG 300L, pectinase가 주성분인 Pectinex 100L 및 cellulase, $\beta$-glucosidase, xylanase가 주성분인 Viscozyme 등을 단독 또는 혼합 처리하여 생리활성이 우수한 무배당체 화합물의 함 량을 증가할 수 있었다. 감귤에 함유되어 있는 flavonoid 화합물의 함량은 naringenin이 $100\∼200\;ng/g$(감귤건조중량), hesperetin이 1$100\∼220\;ng/g$(감귤건조중량) 정도이었으나 당분해효소를 처리한 감귤에 함유되어 있는 활성형 flavonoid 화합물인 naringenin이 $1539\∼6674\;ng/g$(감귤건조중량), hesperetin이 $1974\∼8906\;ng/g$(감귤건조중량)으로서 본 실험 결과를 통해 감귤 추출물의 flavonoid 화합물의 함량을 약 $10\∼80$배 정도로 증가시킬 수 있었다. 당 분해 효소에 의한 무배당체로의 최적 전환조건으로는 당 분해 효소의 $5\%$ 첨가, pH $5\∼7$이었으며 효소 반응 시간으로는 $24\∼48$시간이 가장 우수하였다. 따라서 본 연구에 의해 제조된 감귤 조성물에는 생리활성이 우수한 활성형의 무배당체 flavonoid인 naringenin과 hesperetin을 다량으로 함유하고 있어 기능성 식품, 건강 보조식품, 가공 식품의 식품 원료나 사료 첨가제 의 용도로 유용하게 사용할 수 있다.

• Journal of Microbiology and Biotechnology
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• 제18권3호
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• pp.512-519
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• 2008

Three different polysaccharide-peptide complexes (PPC, named as Fr-I, Fr-II, and Fr-III) were produced by submerged mycelial culture of an entomopathogenic fungus Cordyceps sphecocephala, and their anticancer activities were investigated in human hepatocarcinoma (HepG2) and neuroblastoma (SK-N-SH) cells. The highest inhibitory effects of PPC on both HepG2 and SK-N-SH cells were achieved with Fr-I, whereas Fr-III with low molecular mass showed lower inhibition effects. Interestingly, the inhibitory effects of the three fractions were increased after protease digestion, suggesting that the inhibitory effects resulted mainly from the carbohydrate moiety, at least in the case of Fr-II and Fr-III, of PPC. The results of DNA fragmentation in PPC-induced apoptotic cells were confirmed by both DNA ladder assay and comet assay. Our investigation also showed that PPC-induced apoptosis of both cancer cells was associated with intracellular events including DNA fragmentation, activation of caspase-3, and modulation of Bcl-2 and Bax. We conclude that PPC has potential as a novel therapeutic agent for the treatment of both HepG2 and SK-N-SH cancer cells without any cytotoxicity against normal cells.

• 한국어병학회지
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• 제20권3호
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• pp.307-313
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• 2007

In this study, a QCM biosensor was made to detect Edwardsiella tarda (E. tarda) using a specific antibody. A 9 MHz AT-cut piezoelectric wafer layered with two gold electrodes of 5mm diameter had a reproducibility of 0.1 Hz in frequency response and was used as the transducer of the QCM biosensor. Self assembled layer (SAM) was conformed on a quartz crystal by treating with 3-mer-captopropionic acid (MPA) and activated with N-ethyl-N'-(3-dimethyl-aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS). The resulting NHS group was further converted to hydrazide by the reaction with hydrazine. Aldehyde group was introduced into the carbohydrate moiety of anti-E. tarda antibody by the reaction with periodic acid and was used to immobilise the antibody through the reaction with hydrazide group on the electrode surface. A baseline was established in the presence of phosphate-buffered saline (PBS) and a resonant frequency (F1) was measured. Sample was added to the sensor surface and second resonant frequency (F2) was measured after unbound substances were washed out with PBS several times. Finally, the frequency shift (ΔF) representing the mass change was calculated by subtracting F2 from F1. After adding the oxidized anti-E. tarda antibody to the electrode surface containing hydrazide group, frequency shift of 288.811.4 Hz (mean S.E) was observed, thus proving that considerable amount of antibody was immobilized. In the immunoassay test, the frequency shift of 1877.75 Hz, 580.67 Hz, 221.39 Hz, 7.671.83 Hz (mean S.E) were observed at doses of 1000, 500, 100, 50 g of bacterial cells, respectively. It was also demonstrated that the prepared sensor chip was stable enough to withstand repeated surface regeneration with 0.2 M Tris-glycine and 1 % DMSO, pH 2.3 more than ten times.

• Journal of Microbiology and Biotechnology
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• 제11권1호
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• pp.21-28
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• 2001

Enzyme linked-immunosorbent assay (ELISA) for a rapid detection of fungi, Aspergillus and Penicillium genera in food, were developed and their efficiencies were approved by detecting artificially contaminated agricultural commodities. Mice were immunized with partially purified Aspergillus flavus extracellualr polysaccharide (EPS) and lymph node cells of the mice were fused with the myeloma cells for production of monoclonal antibodies. Mab 1G11, one of the antibodies, was selected and purified. A sandwich ELISA was established and its detection limit toward A. flavus EPS was 1mg/ml. Among the 59 strains tested (including 18 species of Aspergillus, 16 of Penicillium, 11 of Fusarium, 1 of Absidia, 2 of Alternaria, 2 of Candida, 2 of Cladosporium, 2 of Geotrichum, 2 of Mucor, 2 of Rhizopus, 1 of Trichoderma), species of Aspergillus and penicillium had a high reactivity with Mab 1G11 even up to 10,000 times dilution of culture broths. The other genera except Cladosporium resinae showed no reactivity, thus Mab 1G11 was specific to the genera of Aspergillus and Penicillium. The epitope of A. flavus EPS against monoclonal Mab 1G11 was on the carbohydrate moiety when 1 to 100$\mu g/g$ A. flavus EPS were put into rice, potato, and mandarin orange, the average recoveries detected by sandwich ELIA were 123, 59, and 76%, respectively. Correlation was found to be linear between the EPS, and mycelium of A. flavus and Penicillium citrinum grown in a liquid medium (r=0.87 and 0.96), and also between the EPS and colony forming unit in solid media of rice of potato (r=0.91-0.99).

• Journal of Microbiology and Biotechnology
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• 제16권8호
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• pp.1240-1247
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• 2006

The present study investigated the influence of the aeration rate and agitation intensity on the production of the mycelial biomass and antioxidative exopolysaccharide (EPS) in Ganoderma resinaceum. In submerged cultures with varying agitation speeds and aeration rates in a stirred-tank reactor, the maximum mycelial biomass and maximum EPS concentration were achieved at 50 rpm and 300 rpm, respectively. Under varying aeration rates, the highest amount of mycelial biomass (18.1 g/l) was accumulated at the lowest aeration rate (0.5 vvm) and the maximum EPS production (3.0 g/l) obtained at 1.0 vvm. A compositional analysis revealed that the five different EPSs were protein-bound heteropolysaccharides, consisting of 87.17-89.22% carbohydrates and 10.78-12.83% proteins. The culture conditions had a striking affect on the carbohydrate composition of the EPS, resulting in different antioxidative activities. All the EPSs showed strong scavenging activities against superoxide and 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radicals, whereas no clear trend in antioxidative activity was observed against hydroxyl radicals and lipid peroxides. Although the precise reason for this difference is still unclear, the high glucose moiety of EPS is probably linked to its broad spectrum of antioxidative activity.

• Bulletin of the Korean Chemical Society
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• 제32권9호
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• pp.3327-3332
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• 2011

Antimicrobial peptides have recently gained the much attention because of their ability to make defense system from attacking bacterial infections. Drosocin has been considered as very attractive antibiotic agents because of low toxicity against human erythrocytes and active at the low concentration. We have studied the structureactivity relationship of a glycopeptide drosocin focused on the N-acetyl-D-galactoside at $Thr^{11}$ residue. Based on the radial diffusion assay, we found that the acetylation of carbohydrate moiety increased the antimicrobial activity and the $Pro^{10}$, present in the middle of drosocin plays an important role in the antimicrobial activity. Our results provide a good lead compound for further studies on the design of drosocin-based analogues targeting glyco linked Thr site.

• 한국식품영양학회지
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• 제10권3호
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• pp.375-381
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• 1997

식용버섯을 대상으로 항응고 활성을 검색한 결과 영지버섯 알칼리 추출물이 가장 높은 활성을 보였다. 영지버섯으로부터 추출한 조다당 획분인 GL-I은 1N NaOH로 8시간 추출물(GL-0)을 methanol 환류, 에탄올 침전을 거쳐 투석, 동결건조하여 조제하였다. 히-I 획분은 periodate 산화에 의해서는 활성이 크게 변하지만 pronase 소화시 활성의 변화가 거의 없는 것으로 보아 항응고 활성의 본체는 주로 당과 관계되는 것으로 추정된다. 히-I의 구성당은 glucose:galactose:xylose:mannose:arabinose가 19.3:3.0:2.3:1.3:1.0:0.3의 molar ratio로 구성되어 있다. GL-I 획분을 DEAE-Toyopearl 650C(GL-Ia$\longrightarrow$If)와 Sephadex G-100(GL-Ic-I$\longrightarrow$GL-Ic-II)을 이용하여 부분 정제하였다.

• 한국축산식품학회지
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• 제21권4호
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• pp.367-373
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• 2001

MFGM 당단백질의 하나인 PAS-7을 GPC 및 affinity chromatography에 의해 정제하여 2-AB로 형광 표식한 후, anion-exchange column 및 reversed-phase column을 이용해 5개의 성분을 분리하였다. 그 중 가장 상대량이 많은 성분 e에 대하여 RAAM2000을 이용한 당쇄구조 해석을 실시하여, 성분 e는 RAAM2000 GPC에 의하여 4개의 성분으로 분리되어 각각 calibration standard 12.10, 8.88, 5.84 및 4.86GU의 용출위치에 검출되었다. 이 용출위치와 당쇄구조는 livrary의 component-7457과 일치하며, 12.2GU의 크기로 분자량은 1788로 판단되며 library의 당쇄와 약 85%의 확률로 일치했다. 그 결과 성분 e의 당쇄구조는 환원말단에 $\alpha$1-6결합된 fucose를 1개 함유하며, core부분의 비환원말단에 N-acetyllactosamine branch를 2개 함유한 전형적인 biantennary 당쇄구조인 것으로 추축되어, 이전 HPLC, acetolysis, sequential exoglycosidase 소화, NMR분석에 의해 보고된 성분 7N1A의 구조와 일치함으로써, OGS RAAM2000을 이용한 PAS-7의 당쇄구조 해석의 유용성이 증명되었다.

• Journal of Microbiology and Biotechnology
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• 제14권4호
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• pp.658-664
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• 2004

To find a new utilization of rice bran, nine higher fungi were examined for the production of exo-biopolymer with macrophage stimulating activity from rice bran. Among the exo-biopolymers produced from submerged cultures, Lentinus edodes showed the highest activity, followed by Grifola frondosa, Schizophyllum commune, and Coriolus versicolor. L. edodes also had the most potent macrophage stimulating activity in a liquid culture rather than in a solid culture. In order to improve rice bran utilization and the yield of exo-biopolymer with macrophage stimulating activity, the treatment of Rapidase effectively increased the macrophage stimulating activity (about 30% increase), whereas the other enzymes (Econase, Viscozyme, Ultraflo, Celluclast, and Thermylase) treatments did not increase the macrophage stimulating activity. Exo-biopolymer with macrophage stimulating activity from L. edodes contained mainly neutral sugars (58.7%) with considerable amounts of uronic acid (32.2%) and a small amount of proteins (9.1%). Component sugars of exo-biopolymer consisted of mainly arabinose, galactose, glucose, mannose, and xylose (0.95:0.81:0.96:1.00:0.39, respectively). When the exo-biopolymer was treated with $NaIO_4, NaClO_2$, and pronase, the $NaClO_2$ treatment and pronase digestion had little effect, whereas $NaIO_4$ oxidation significantly decreased the macrophage stimulating activity (47.6% reduction at $100\mug/ml$). Therefore, the carbohydrate moiety in exo-biopolymer from L. edodes plays an important role in the expression of the macrophage stimulating activity.