• Journal of Microbiology and Biotechnology
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• 제26권6호
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• pp.1087-1097
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• 2016

An intracellular lipase from Anoxybacillus flavithermus HBB 134 was purified to 7.4-fold. The molecular mass of the enzyme was found to be about 64 kDa. The maximum activity of the enzyme was at pH 9.0 and 50℃. The enzyme was stable between pH 6.0 and 11.0 at 25℃, 40℃, and 50℃ for 24 h. The K_m and V_max of the enzyme for pNPL substrate were determined as 0.084 mM and 500 U/mg, respectively. Glycerol, sorbitol, and mannitol enhanced the enzyme thermostability. The enzyme was found to be highly stable against acetone, ethyl acetate, and diethyl ether. The presence of PMSF, NBS, DTT and β-mercaptoethanol inhibited the enzyme activity. Hg²⁺, Fe³⁺, Pb²⁺, Al³⁺, and Zn²⁺ strongly inhibited the enzyme whereas Li⁺, Na⁺, K⁺, and NH₄⁺ slightly activated it. At least 60% of the enzyme activity and stability were retained against sodium deoxycholate, sodium taurocholate, n-octyl-β-D-glucopyranoside, and CHAPS. The presence of 1% Triton X-100 caused about 34% increase in the enzyme activity. The enzyme is thought to be a true lipase since it has preferred the long-chain triacylglycerols. The lipase of HBB 134 cleaved triolein at the 1- or 3-position.

• Journal of Microbiology and Biotechnology
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• 제22권12호
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• pp.1731-1739
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• 2012

In this study, seven strains isolated from mustard leaf kimchi were screened for their tolerance to simulated gastric and bile juices, the adhesive properties to Caco-2 cells, and the inhibition ability of Salmonella Typhimurium ATCC 29631 adhesion. Lactobacillus acidophilus GK20, Lactobacillus paracasei GK74, and Lactobacillus plantarum GK81, which were resistant to bile as well as gastric juices, possessed high bile-salt hydrolase (BSH) activity towards both sodium glycocholate and sodium taurocholate. The strongest in vitro adherence of $53.96{\pm}4.49%$ was exhibited by L. plantarum GK81 followed by L. acidophilus GK20 with adhesion levels of $40.72{\pm}9.46%$. The adhesion of these strains was significantly (p < 0.05) reduced after exposure to pepsin and heating for 30 min at $80^{\circ}C$. Addition of $Ca^{2+}$ led to a significant (p < 0.05) increase of the adhesion of L. acidophilus GK20, but the adhesion ability of L. plantarum GK81 was not different from the control by the addition of calcium. In the competition and exclusion experiment, the adhesion inhibition of S. Typhimurium by L. plantarum GK81 strain was much higher than the other strains. Moreover, the exclusion inhibition of S. Typhimurium by L. acidophilus GK20 was considerably high, although the inhibition activity of this strain was lower than L. plantarum GK81.

• 한국축산식품학회지
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• 제32권4호
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• pp.409-413
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• 2012

This study aimed to compare the probiotic characteristics of twelve strains of Lactobacillus acidophilus including cholesterol assimilation and conjugated linoleic acid (CLA) production. Cholesterol assimilation exhibited some variation among L. acidophilus strains, which could be classified into three groups based on their assimilation levels (p<0.05). The high cholesterol assimilation group exhibited a significantly higher tolerance to 0.3 and 0.5% bile acid than the low cholesterol assimilation group (p<0.05). Cholesterol assimilation showed positive correlation with 0.5% bile tolerance, and a negative correlation with acid tolerance (p<0.01). Glycocholate deconjugation activity showed no relationship with cholesterol assimilation, whereas taurocholate deconjugation activity was shown to have negative correlation with cholesterol assimilation (p<0.05). CLA production by L. acidophilus strains exhibited a wide variation, ranging from 2.69 to 5.04 mg/g fat. CLA production of L. acidophilus GP1B was the highest among the tested strains, but there was no evidence for differences in CLA production in strain specificity. Based on these results, the cholesterol assimilation of L. acidophilus strains may not be related to deconjugation activity, but may in-fact be attributed to their bile-tolerance.

• Parasites, Hosts and Diseases
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• 제61권4호
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• pp.428-438
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• 2023

Clonorchis sinensis is commonly found in East Asian countries. Clonorchiasis is prevalent in these countries and can lead to various clinical symptoms. In this study, we used overlap extension polymerase chain reaction (PCR) and the Xenopus laevis oocyte expression system to isolate a cDNA encoding the choline transporter of C. sinensis (CsChT). We subsequently characterized recombinant CsChT. Expression of CsChT in X. laevis oocytes enabled efficient transport of radiolabeled choline, with no detectable uptake of arginine, α-ketoglutarate, p-aminohippurate, taurocholate, and estrone sulfate. Influx and efflux experiments showed that CsChT-mediated choline uptake was time- and sodium-dependent, with no exchange properties. Concentration-dependent analyses of revealed saturable kinetics consistent with the Michaelis-Menten equation, while nonlinear regression analyses revealed a Km value of 8.3 µM and a Vmax of 61.0 pmol/oocyte/h. These findings contribute to widen our understanding of CsChT transport properties and the cascade of choline metabolisms within C. sinensis.

• Preventive Nutrition and Food Science
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• 제16권2호
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• pp.104-109
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• 2011

Recently, we have demonstrated that green tea extract (GTE) decreases the intestinal absorption of benzo[a]pyrene (BAP), which is an extremely lipophilic food contaminant. The present study was conducted to examine if an enteral infusion of GTE would influence the biliary secretion of BAP and lipids in rats. Female rats were fed an AIN-93G diet with or without (control) GTE at 5 g/kg diet for 4 week. Following the 4-week dietary treatment, rats with bile duct cannula were infused continuously for 8 hr at 3.0 mL/hr via a duodenal catheter with a lipid emulsion containing $4.0\;{\mu}mol$ BAP labeled with $^{14}C$ ($^{14}C$-BAP), $20.7\;{\mu}mol$ cholesterol, $452\;{\mu}mol$ triolein, and $3.1\;{\mu}mol$ ${\alpha}$-tocopherol, and $396.0\;{\mu}mol$ Na-taurocholate with or without 76.1 mg GTE powder in PBS buffer (pH, 6.4). Bile was collected hourly via bile cannula for an 8 hr period. Our results showed that bile flow did not differ between groups. However, the biliary secretion of $^{14}C$-BAP was significantly enhanced by GTE infusion, compared with those infused with the lipid emulsion alone. However, GTE did not affect the biliary outputs of cholesterol, fat, phospholipid and ${\alpha}$-tocopherol. These findings indicate that GTE has a profound stimulatory effect on the biliary excretion of BAP in rats, without affecting other biliary lipids. The mechanism(s) by which GTE enhances the biliary secretion of BAP remains to be investigated.

• Preventive Nutrition and Food Science
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• 제13권2호
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• pp.71-76
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• 2008

This study was performed to examine the effects of onion powder supplementation on lipid metabolism in male SD rats fed a high fat and high cholesterol diet. Experimental groups were control (C), high fat fed group (HF), high fat+onion powder intake (OP), high fat+quercetin intake (Q). The HF diet contained 1% cholesterol, 4% lard, 0.3% Na-taurocholate, and quercetin supplementation level was 0.1 g/kg diet. The OP group showed lower body weight gains compared to the control, while there was no significant difference in food efficiency ratio efficiency. When the proportion of fecal bile acids per total lipids was calculated, there was a significant decrease in the HF group compared to the control group, while the levels of the control group was same as that of the OP group. There was no significant difference between the HF and Q groups in bile acid/total lipid in feces. The amount of total cholesterol in liver increased significantly in HF group compared to the control group, while total cholesterol decreased significantly in the OP group compared to the HF group. There was a significant decrease in GOT (glutamic oxaloacetic transaminase) activity in OP and Q groups compared to the HF group. In conclusion, feeding onion powder to hyperlipidemic rates appeared to control weight gain, significantly lower the level of total cholesterol in the liver, and recover GOT activity. We also demonstrated that onion powder intake was more effective than quercetin intake.

• The Korean Journal of Physiology and Pharmacology
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• 제19권4호
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• pp.299-307
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• 2015

Severe acute pancreatitis (SAP) is normally related to multiorgan dysfunction and local complications. Studies have found that local pancreatic renin-angiotensin system (RAS) was significantly upregulated in drug-induced SAP. The present study aimed to investigate the effects of angiotensin II receptors inhibitor valsartan on dual role of RAS in SAP in a rat model and to elucidate the underlying mechanisms. 3.8% sodium taurocholate (1 ml/kg) was injected to the pancreatic capsule in order for pancreatitis induction. Rats in the sham group were injected with normal saline in identical locations. We also investigated the regulation of experimentally induced SAP on local RAS expression in the pancreas through determination of the activities of serum amylase, lipase and myeloperoxidase, histological and biochemical analysis, radioimmunoassay, fluorescence quantitative PCR and Western blot analysis. The results indicated that valsartan could effectively suppress the local RAS to protect against experimental acute pancreatitis through inhibition of microcirculation disturbances and inflammation. The results suggest that pancreatic RAS plays a critical role in the regulation of pancreatic functions and demonstrates application potential as AT1 receptor antagonists. Moreover, other RAS inhibitors could be a new therapeutic target in acute pancreatitis.

• 한국식품영양과학회지
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• 제45권6호
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• pp.783-788
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• 2016

본 연구는 수수의 섭취가 대표적인 트랜스지방산인 elaidic acid, linoelaidic acid와 cholesterol의 소장흡수에 어떤 영향을 미치는지를 조사하기 위하여 설계하였다. 소장의 지방흡수를 in vivo 상태에서 측정하기 위해 소장흡수율 측정모델을 이용하여 십이지장주입관과 림프채취관을 삽입하는 미세수술을 진행하였다. 실험동물의 십이지장으로 $146.4{\mu}mol$ trielaidin, $36.8{\mu}mol$ trilinoelaidin, $452.0{\mu}mol$ triolein, $1.0{\mu}Ci$ $^{14}C-cholesterol$, $20.7{\mu}mol$ cholesterol, $396.0{\mu}mol$ Na-taurocholate, PBS buffer(pH 6.4)가 포함된 지질유화액만 공급받는 동물군을 대조군, 수수추출물이 혼합된 지질유화액을 공급받는 동물군을 수수군으로 설계하였다. 지질유화액은 시간당 3 mL씩 8시간 동안 주입하면서 림프관으로 흡수되는 elaidic acid, linoelaidic acid와 각종 지방산 그리고 $^{14}C-cholesterol$의 양을 분석하여 수수에 의한 영향을 비교하였다. 먼저 8시간 동안 분비된 림프액의 양은 대조군과 수수군 간에 유의적인 차이를 나타내지 않았다. 하지만 8시간 동안의 elaidic acid 흡수는 수수추출물의 공급으로 유의적으로 억제되는 것으로 나타났다. Linoelaidic acid 또한 마찬가지로 수수추출물 공급으로 유의적으로 그 흡수가 억제된 것으로 나타났다. 트랜스지방산을 비롯해 palmitic acid, stearic acid, oleic acid, linoleic acid, arachidonic acid, docosahexaenoic acid, $^{14}C-cholesterol$ 또한 대조군보다 수수군에서 흡수가 유의적으로 억제된 것으로 나타났다. 이상의 결과들은 수수추출물이 트랜스지방산과 cholesterol의 소장 흡수를 억제할 수 있다는 사실을 증명한다. 이번 연구를 바탕으로 수수 섭취가 트랜스지방이나 cholesterol의 흡수를 농도 비례적으로 억제할 수 있을지, 소화흡수 과정 중 정확히 어떤 과정에 영향을 미쳤는지에 관한 연구가 앞으로 진행될 과제라고 판단된다.

• 한국식품저장유통학회지
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• 제15권3호
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• pp.450-455
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• 2008

이 실험은 녹차추출물이 흰쥐 소장mucosa에서 $^{14}C$-oleic acid의 에스테르화 과정에 어떤 영향을 미치는지를 조사하기 위해서 설계되었다 먼저, olive oil을 구강으로 강제 투여했을 때, 1시간 그리고 5시간 후에, 녹차 성분의 변수에 의해서 소장 mucosa 내에 축적된 중성지방이 얼마나 감소하는지를 조사하였다. 그리고 소장세포 내에서 녹차 성분에 의해서 각종 지방으로 $^{14}C$-oleic acid의 분배비율이 영향을 받는지 조사하였다. 이 실험에 사용된 micelle 용액은 $200.0\;{\mu}Ci$ $^{14}C$-oleic acid, $200.1\;{\mu}mol$ oleic acid, $200.1\;{\mu}mol$ 2-monooleoylglycerol, $66.7\;{\mu}mol$ palmitoyl-sn-glycero-3-phosphocholine, 2.2 mmol glucose, $50.0\;{\mu}mol$ albumin, 그리고 16.5 mmol Na-taurocholate per L of phosphate buffered saline (pH, 6.3)를 기본으로 하며, 실험군의 micelle 용액은 녹차추출물 분말(green tea extract powder)이 8.87 g/L의 농도로 추가되었다. Ligament of Treitz 부위 에 연결된 소장 부위의 양 끝을 살균 처리된 suture silk(4-0 Silk)로 느슨하게 매듭처리 한 후, $800.0\;{\mu}L$ micelle 용액을 매듭 처리된 소장의 윗매듭 속으로 주입하여, 장기 및 부속기관을 원위치시켰다. 10분 후에, cervical dislocation 방법으로 희생시킨 후 매듭된 소장 부위를 꺼내어 장내용물(luminal content)의 잔존 $^{14}C$-oleic acid를 측정하였다. 획득된 소장 벽에서 각종 지방으로 $^{14}C$-oleic acid의 분배율을 측정하였다. 소장 mucosa에서 총지방량은 녹차추출물에 의해서 영향을 받지 않았으나, 이중 중성지방의 비율은 유의적으로 감소되었다. 또한 소장 벽에서 각종지방으로 $^{14}C$-oleic acid의 분배율은 녹차추출물에 의해서, 중성지방과 인지질 분획에서 유의적인 감소현상을 보였다. 이 실험은, 이전 동물실험들에서 누차 보고되었던, 녹차추출물에 의한 소장 지방 흡수 억제 현상은 $^{14}C$-oleic acid의 에스테르화 비율과 상관성이 있는 것으로 제시하고 있다.

• Journal of Pharmaceutical Investigation
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• 제39권2호
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• pp.97-106
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• 2009

Although lovastatin (LS) is widely used in the treatment of hypercholesterolemia, its bioavailability is known to be around 5%. This study was aimed to increase the solubility and dissolution-permeation rates of LS using solid dispersions (SDs) with bile salts. The solubilities of LS in water, aqueous bile salt solutions and non-aqueous vehicles were determined, and effects of bile salts on the cellulose or duodenal permeation of LS from SDs were evaluated using a horizontal permeation system. SDs were prepared at various ratios of LS to carriers, such as sodium deoxycholate (SDC), sodium glycocholate (SGC) and/or 2-hydroxypropyl-$\beta$-cyclodextrin (HPCD). The addition of bile salts (25 mM) in water increased markedly the solubility of LS by the micellar solubilization. Some non-aqueous vehicles were effective in solubilizing LS. From differential scanning calorimetric studies, it was found that the crystallinity of LS in SDs disappeared, indicating a formation of amorphous state. The SDs showed markedly enhanced dissolution compared with those of their physical mixtures (PMs) and drug alone. In the dissolution-permeation studies using a cellulose membrane, the donor and receptor solutions were maintained as a sink condition using pH 7.0 phosphate buffer containing 0.05% sodium lauryl sulfate (SLS). The flux of LS alone was nearly same as that of LS-SDC-HPCD (1:3:6) PM. However, the flux of LS-SDC-HPCD (1:3:6) SD slightly increased compared with drug alone and PM, suggesting that entrapment of LS in micelles does not significantly hinder the permeation across cellulose membrane. In the dissolution-duodenal permeation studies using a LS-HPCD-SDC (1:3:6) SD, the addition of various bile salts in donor solutions (25 mM) enhanced the permeation of LS markedly, and the fluxes were found to be $0.69{\pm}0.41$, $0.87{\pm}0.51$, $0.84{\pm}0.46$, $0.47{\pm}0.17$ and $0.68{\pm}0.32{\mu}g/cm^2/hr$ for sodium cholate (SC), SDC, SGC, sodium taurodeoxycholate (STDC) and sodium taurocholate (STC), respectively. The stepwise increase of donor SGC concentration increased the flux dose-dependently. From the relationship of donor SGC concentration and flux, the concentration of SGC initiating the permeation across the duodenal mucosa was calculated to be 11.1 mM, which is nearly same as the critical micelle concentration (CMC, 11.6 mM) of SGC. However, with no addition of bile salts and below CMC, the permeation was very limited and irratic, indicating that LS itself is very poor permeable. Higher protions of bile salt in SD such as LS-SDC or LS-SGC (1 : 49 and 1 : 69) showed highly promoted fluxes. In conclusion, SD systems with bile salts, which may form their micelles in intestinal fluids, might be a promising means for providing enhanced dissolution and intestinal permeation of practically insoluble and non-absorbable LS.