• 한국식품과학회지
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• 제29권5호
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• pp.1067-1070
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• 1997

Chelating sepharose fast flow gel에 $Cu^{2+}$을 고정화시키고 이들과 단백질과의 친화력 정도에 의해서 용출용매를 통해 단백질을 분획하는 방식의 IMAC법을 이용하여 소의 혈장단백질로부터 IgG의 분리를 시도하였다. 대부분의 혈장단백질은 1차(0.01 M $Na_2HPO_4$, 0.5 M NaCl, pH 4.0)와 2차 용출용매(0.01 M imidazol)에 의하여 용출되었으며 역상 chromatography를 이용하여 각 분획의 단백질조성을 조사한 결과, 1차 용출용매에 의해서는 주로 albumin이, 그리고 2차 용출용매에 의해서는 IgG와 transferrin 등이 IMAC column으로부터 용출되었다. 2차 용출용매에 의해 얻어진 분획으로부터 분자량 100 kD이상의 단백질을 한외여과장치를 이용하여 농축하였을 때, IgG가 효과적으로 분리 정제 되었다.

• Food Science and Biotechnology
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• 제15권1호
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• pp.24-27
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• 2006

A simple HPLC quantification method was developed for genistein, genistin, daidzein, and daidzin in soybeans and soybean products. The procedure used a $4.6{\times}100\;mm$ $Chromolith^{(R)}$ RP-18e column with a mobile phase of 1% HOAc in 20% MeOH to 1% HOAc in 80% MeOH for 10 min. The injection volume was $2\;{\mu}L$ at a flow rate of 2 mL/min. Detection was carried out under UV at 254 nm. Under these conditions, the major isoflavones daidzein, daidzin, genistein, and genistin in soybean and soybean pastes were eluted within 7 min with baseline separation. Optimal extraction of the above four major isoflavones was achieved when 40 g of soybean or soybean paste was refluxed in 100 mL of 95% ethanol for 2 hr. Ten different soybean cultivars and nine commercial soybean pastes were analyzed by this method. The total isoflavone content was highest in the cultivar Somyung ($2,497\;{\mu}g/g$ dry weight). The isoflavone content in soybean pastes varied widely from manufacturer to manufacturer (an almost five-fold difference between the highest and lowest values). Such variations were presumably due to differences in fermentation conditions, type of soybeans used, and levels of such additives as starch and salt.

• Journal of Microbiology and Biotechnology
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• 제24권4호
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• pp.465-474
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• 2014

This study was performed to investigate the inhibitory effects of brown algae extracts on histamine production in mackerel muscle. First, antimicrobial activities of brown algae extracts against Morganella morganii were investigated using a disk diffusion method. An ethanol extract of Ecklonia cava (ECEE) exhibited strong antimicrobial activity. The minimum inhibitory concentration (MIC) of ECEE was 2 mg/ml. Furthermore, the brown algae extracts were examined for their ability to inhibit crude histidine decarboxylase (HDC) of M. morganii. The ethanol extract of Eisenia bicyclis (EBEE) and ECEE exhibited significant inhibitory activities (19.82% and 33.79%, respectively) at a concentration of 1 mg/ml. To obtain the phlorotannin dieckol, ECEE and EBEE were subjected to liquid-liquid extraction, silica gel column chromatography, and HPLC. Dieckol exhibited substantial inhibitory activity with an $IC_{50}$ value of 0.61 mg/ml, and exhibited competitive inhibition. These extracts were also tested on mackerel muscle. The viable cell counts and histamine production in mackerel muscle inoculated with M. morganii treated with ${\geq}2.5 $ MIC of ECEE (weight basis) were highly inhibited compared with the untreated sample. Furthermore, treatment of crude HD-Cinoculated mackerel muscle with 0.5% ECEE and 0.5% EBEE (weight basis), which exhibited excellent inhibitory activities against crude HDC, reduced the overall histamine production by 46.29% and 56.89%, respectively, compared with the untreated sample. Thus, these inhibitory effects of ECEE and EBEE should be helpful in enhancing the safety of mackerel by suppressing histamine production in this fish species.

• Journal of Microbiology and Biotechnology
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• 제23권2호
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• pp.167-176
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• 2013

A novel bioactive molecule produced by Bacillus thuringiensis subsp. kurstaki Bn1 (Bt-Bn1), isolated from a common pest of hazelnut, Balaninus nucum L. (Coleoptera: Curculionidae), was determined, purified, and characterized in this study. The Bt-Bn1 strain was investigated for antibacterial activity with an agar spot assay and well diffusion assay against B. cereus, B. weinhenstephenensis, L. monocytogenes, P. savastanoi, P. syringae, P. lemoignei, and many other B. thuringiensis strains. The production of bioactive molecule was determined at the early logarithmic phase in the growth cycle of strain Bt-Bn1 and its production continued until the beginning of the stationary phase. The mode of action of this molecule displayed bacteriocidal or bacteriolytic effect depending on the concentration. The bioactive molecule was purified 78-fold from the bacteria supernatant with ammonium sulfate precipitation, dialysis, ultrafiltration, gel filtration chromatography, and HPLC, respectively. The molecular mass of this molecule was estimated via SDS-PAGE and confirmed by the ESI-TOFMS as 3,139 Da. The bioactive molecule was also determined to be a heat-stable, pH-stable (range 6-8), and proteinase K sensitive antibacterial peptide, similar to bacteriocins. Based on all characteristics determined in this study, the purified bacteriocin was named as thuricin Bn1 because of the similarities to the previously identified thuricin-like bacteriocin produced by the various B. thuringiensis strains. Plasmid elution studies showed that gene responsible for the production of thuricin Bn1 is located on the chromosome of Bt-Bn1. Therefore, it is a novel bacteriocin and the first recorded one produced by an insect originated bacterium. It has potential usage for the control of many different pathogenic and spoilage bacteria in the food industry, agriculture, and various other areas.

• Journal of Microbiology and Biotechnology
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• 제27권7호
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• pp.1223-1232
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• 2017

Cordyceps militaris, a member of Ascomycota, a mushroom referred to as caterpillar Dong-chung-ha-cho, is commercially valuable because of its high content of bioactive substances, including cordycepin, and its potential for artificial cultivation. Cordycepin (3'-deoxyadenosine) is highly associated with the pharmacological effects of C. militaris. C. militaris is heterothallic in that two mating-type loci, idiomorph MAT1-1 and MAT1-2, exist discretely in two different spores. In this study, nine C. militaris strains were mated with each other to prepare newly bred strains that produced a larger amount of cordycepin than the parent strains. Nine strains of C. militaris were identified by comparing the internal transcribed spacer sequence, and a total of 12 single spores were isolated from the nine strains of C. militaris. After the MAT idiomorph was confirmed by PCR, 36 mating combinations were performed with six single spores with MAT1-1 and the others with MAT1-2. Eight mating combinations were successfully mated, producing stroma with perithecia. Cordycepin content analysis of all strains by high-performance liquid chromatography revealed that the KASP4-bred strain produced the maximum cordycepin among all strains, regardless of the medium and stroma parts. Finally, universal rice primer-PCR was performed to demonstrate that the bred strains were genetically different from the parental strains and new C. militaris strains. These results may be related to the recombination of genes during mating. The newly produced strains can be used to meet the industrial demand for cordycepin. In addition, breeding through mating suggests the possibility of producing numerous cordycepin-producing C. militaris strains.

• 한국식품과학회지
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• 제29권3호
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• pp.470-475
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• 1997

식혜 제조시 쌀 품종에 따른 당도의 변화를 조사하기 위하여 45종의 쌀 품종을 사용하였을 때 당화 10시간 후의 당도가 $9.0{\sim}11.1\;Brix$로 품종간에 차이를 나타내었다. 간척과 신금오, 서안, 계화 등을 이용시 당도가 높게 나타났으며, 상주, 남원, 영덕, 오봉 등은 당도가 상대적으로 낮았다. 간척과 상주품종을 비교하면 식혜제조시 당화 10시간에 약 19%의 당도 차이가 있었다. 쌀 품종 간척을 이용한 식혜의 HPLC분석에 의한 주요 당 조성은 fructose 3.6%, glucose 9.8%, maltose 78.3%, maltotriose 8.3%이었다. 육조 맥아와 이조 맥아를 이용한 당화력의 차이는 육조 맥아를 100 메쉬로 분쇄하여 사용시 이조 맥아보다 높았다. 육조 맥아의 당화 최적온도는 $60^{\circ}C$이었으며, 밥과 맥아 추출물의 비를 1:8로 하였을 때 맥아 첨가량 25% 추출물에서 당화력이 가장 높았다.

• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 2003년도 정기총회 및 학술발표회
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• pp.51-51
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• 2003

The medicinal plant Cimicifuga Racemosa (Black cohosh) has been used to treat many kinds of neuronal and menopausal symptoms, such as arthritis, menopausal depression, nerve pain, etc. Here, we examined the effect of Cimicifugoside (CF), one of triterpene glycosides which have been known as pharmacologically active ingredients of C. Racemosa, on nicotinic acetylcholine receptor (nAChR)-mediated catecholamine (CA) secretion in bovine adrenal chromaffin cell. Cimicifugoside inhibited calcium increase induced by 1,1-dimethyl-4-phenylpiperazinium iodide (DMPP), a nAChR agonist with a half maximal inhibitory concentration (IC50) of 18${\pm}$2${\mu}$M. In contrast, cimicifugoside did not affect the calcium increases evoked by high K$\^$+/, veratridine, and bradykinin. The DMPP-induced sodium increase was also inhibited by cimicifugoside with IC50 of 2${\pm}$0.3${\mu}$M, suggesting that the activity of nAChRs is inhibited by cimicifugoside. Cimicifugoside did not effect on the KCl-induced secretion but markedly inhibited the DMPP-induced catecholamine secretion which was monitored by carbon-fiber amperometry in real time, and by high performance liquid chromatography (HPLC) through electrochemical detection. The results suggest that cimicifugoside selectively inhibits nAChR-mediated response in bovine chromaffin cells.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 춘계학술발표회
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• pp.62-62
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• 2018

Inhibition of aldose reductase (AR) has been shown to prevent the onset and progression of many diabetic complications wherein several AR inhibitors were isolated from plants abundant in polyphenolic compounds. Lespedeza cuneata (Fabaceae), a perennial plant indigenous in East Asian countries, is shown to be abundant in these polyphenolic substances such as flavonoids and tannins. However, there are no studies to date regarding its effects on AR. In this study, the inhibitory activity of the methanol extract and stepwise polarity fractions of Lespedeza cuneata on AR was investigated. The bioactive compounds purified from L. cuneata by repeated column chromatography were also tested for AR inhibition. Results show that the ethyl acetate and n-butanol fractions of L. cuneata exhibited potent inhibition against AR with $IC_{50}$ values of 0.57 and $0.49{\mu}g/mL$, respectively. Further analysis led to the isolation of acacetin (1), afzelin (2), astragalin (3), kaempferol (4), and scutellarein 7-O-glucoside (5). The AR inhibitory effects these five compounds were also determined in which compounds 2, 3, and 5 showed potent AR inhibitory effects with $IC_{50}$ values of 2.20, 1.91, and $12.87{\mu}M$, respectively. Quantitative analysis of astragalin (3) by HPLC-UV was also performed in the leaves and roots of L. cuneata (0.626 and 0.671 mg/g, respectively). This study reports that the flavonoids isolated from L. cuneata show promising AR inhibitory activities which can be further developed as natural therapies for treating and managing diabetic complications.

• 약학회지
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• 제42권5호
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• pp.473-479
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• 1998

Lovastatin (LOVA), a fungal metabolite isolated from cultures of Aspergillus terreus, is a competitive HMG-CoA reductase inhibitor used for the treatment of primary hyper cholesterolemia, and has also been shown to suppress growth in a variety of non-glioma tumor cell lines. A sensitive reversed-phase high-perfonnance liquid chromatographic method with ultraviolet (UV) absorbance detection has been developed to quantitate LOVA in human plasma and urine samples using liquid-liquid extraction procedure. Baseline separation of LOVA and internal standard, simvastatin was achieved on a Novapak $C_{18}$ analytical column with a mobile phase containing 0.025M $NaH_2PO_4$: CAN (35:65, v/v%), adjusted pH to 4.5. The flow rate was set at 1.5ml/min, and the column effluent was monitored by a UV detection at 238nm. The limit of quantification was determined to be 0.5${\mu}$g/ml while extraction efficiency of LOVA ranged from 73.4-82.9% at LOVA concentrations of 0.5 to 10${\mu}$g/ml. Good linearity with correlation coefficients greater than 0.999 was obtained in the range of LOVA concentrations from 0.5 to 10${\mu}$g/ml. The accuracy and the precision were proven excellent with relative standard deviation (RSD, %) and relative error (RE, %) of less than 4.2 and 4.0, respectively. Intraday precision, evaluated at five LOVA concentrations (0.5, 1, 2, 5, 10${\mu}$g/ml) and expressed as RSD ranged from 0-1.82% while the interday precision at the same concentrations ranged from 0.7-10.5%. The analytical method described was then successfully employed for the determination of LOVA concentrations in plasma samples obtained during a phase II clinical trial using high doses of LOVA (30-40mg/kg/day). This method could be further utilized for the ongoing pharmacolkinetic studies and therapeutic drug monitoring of the high-dose LOVA therapy in adenocarcinoma patients.

• 한국한의학연구원논문집
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• 제15권2호
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• pp.119-124
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• 2009

The purpose of this study was investigation of quantitative analysis of marker substances in Paeonia lactiflora extracts by solid fermentation. High performance liquid chromatography (HPLC) for the determination of albiflorin and paeoniflorin in P. lactiflora extracts by solid fermentation, the separation method was performed on C18 column ($250\;mm\;{\times}\;4.6\;mm$, $5\;{\mu}m$, RS tech) using gradient solvent mixtures of water-acetonitrile with photodiode array detector (230nm). The flow rate was 1.0 ml/min. Retention time of albiflorin and paeoniflorin was about 28.88, 31.92 min and linearity of calibration was showed good result(r2 = 0.9998, 0.9996), respectively. Content of albiflorin was $0.090\;{\pm}\;0.03%$ in P. lactiflora extract(control), $0.102\;{\pm}\;0.00%$ in P. lactiflora extract fermented with Paecilomyces japonica, $0.056\;{\pm}\;0.01%$ in P. lactiflora extract fermented with Ganoderma lucidum, $0.093\;{\pm}\;0.00%$ in P. lactiflora extract fermented with honey and $0.046\;{\pm}\;0.00%$ in P. lactiflora extract fermented with Nuruk. Content of paeoniflorin was $4.506\;{\pm}\;0.13%$ in control, $2.599\;{\pm}\;0.04%$ in P. lactiflora extract fermented with Paecilomyces japonica, $1.222\;{\pm}\;0.03%$ in P. lactiflora extract fermented with Ganoderma lucidum, $2.750\;{\pm}\;0.05%$ in P. lactiflora extract fermented with honey and $0.847\;{\pm}\;0.00%$ in P. lactiflora extract fermented with Nuruk, respectively. Content of the marker substances did not increase in all fermentation experiment group.