• Journal of Microbiology and Biotechnology
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• 제9권6호
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• pp.832-838
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• 1999

Two different types of lipases (lipase I and lipase II) secreted into culture medium by Rhizopus sp. L-I were purified using a hydrophobic chromatography and were partially characterized. Both enzymes were monomeric as revealed by SDS-PAGE and gel filtration. The molecular masses of the enzymes were identified as 45 kDa (lipase I) and 69 kDa (lipase II). The isoelectric points were estimated to be 3.6 and 5.2 for lipase I and lipase II, respectively. pH and temperature activity optima for lipase I were as 7.5 and $50^{\circ}C$, respectively, whereas the corresponding parameters for lipase II were 6.0 and $45^{\circ}C$. The amino terminal sequences of lipase I and lipase II, determined by Edman degradation, were found to be Leu-Val-Met-Ile-Gln-Arg and Leu-Val-Met-Lys-Gln-Arg, respectively. By western blotting analysis, the two lipases were found to have a common antigenic determinant. Immuno-electron cytochemistry conducted with polyclonal anti-lipase I antibody indicated the enzyme located in both the periplasm and the adjacent vesicles of fungal hyphae. Fortunately, the sites on the cell envelope where lipase was exported into the culture medium was also identified.

• 한국미생물·생명공학회지
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• 제24권1호
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• pp.87-91
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• 1996

The optimum temperature and pH for the enzyme activity were 45$^{\circ}C$ and 10.0, respectively. The enzyme was stable in a pH range of 5 to 10, and 62% of its activity was lost on heat treatment of 60$^{\circ}C$ for 20 min. The activity of the purified enzyme was inhibited by $Fe^{2+},\;Zn^{2+}\;and\;Pb^{2+}$, and slightly activated by $Mn^{2+}\;and\;Ca^{2+}$. ${\gamma}$-Chloromercuribenzoic acid, 2,4-dinitrophenol and $H_{2}O_{2}$ did not show inhibitroy effect on the lipolytic activity of the alkaline lipase but ethylenediaminetetraacetic acid inhibited the enzyem activity. This suggested that the enzyme have metal group in its active site. Sodium salts of bile acids stimulated the enzyme activity. Analysis of hydrolyzates of olive oil after the reaction revealed that Serratia liquefaciens AL-11 produced non-specific lipolytic enzyme.

• 한국식품저장유통학회지
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• 제17권5호
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• pp.727-732
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• 2010

신선한 산양삼 잎을 EtOH로 침지 추출하여 얻어진 추출물을 n-hexane, EtOAc, n-BuOH 로 용매 분획하였다. 이중 pancreatic lipase 저해활성이 상대적으로 높은 EtOAc 분획에 대해 silica 및 $C_{18}$ column chromatography를 이용하여 3개의 flavonoid 화합물을 분리하였다. 각 화합물의 화학구조는 NMR 스펙트럼 데이터 해석 및 표품과의 HPLC 직접 비교를 통하여 kaempferol-3-O-sophoroside (I), astragalin (II), kaempferol (III)로 동정하였다. 이들 화합물중 kaempferol (III)는 $IC_{50}$ 값이 $9.1{\pm}1.5$ mM로 가장 강한 효능을 나타내었으며, 다음으로 C-3 번 위치에 1개의 glucose가 결합한 astragalin (II)의 $IC_{50}$ 값이 $17.4{\pm}2.7$ ${\mu}M$, C-3 번 위치에 2개의 glucose가 결합한 kaempferol-3-O-sophoroside (I)의 $IC_{50}$ 값이 $20.3{\pm}2.2$ ${\mu}M$의 저해능을 나타내었다. 산양삼 잎의 EtOAc 가용부에에 존재하는 flavonoid계 pancreatic lipase 저해활성 물질을 동정하였으며, 이들 활성은 kaempferol 타입의 화합물은 C-3위치에 당 결합에 의한 hydrophobicity와 관련됨을 시사하였다. 향후 이들 활성물질의 활성 기작에 대한 연구가 필요하며 본 연구결과는 보다 우수한 pancreatic lipase 저해능을 가지는 새로운 선도화합물 발굴을 위한 기초자료로 이용될 수 있을 뿐만 아니라 산양삼잎의 식물 화학적 성분에 대한 기초자료로 이용될 수 있을 것으로 사료된다.

• 생명과학회지
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• 제18권8호
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• pp.1159-1163
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• 2008

이 연구의 목적은 물-오일 계면에서 특이적인 lipase 활성을 측정할 수 있는 high-throughput assay 방법을 확립하는 것이다. 이 방법은 pH에 따라 형광의 세기가 변하는 fluorescein의 특성을 이용하여 lipase의 작용으로 방출되는 지방산으로 인한 pH 변화를 fluorescein의 형광 변화로 측정하도록 되어 있다. 활성의 측정은 오일 에멀션과 fluorescein 그리고 효소를 포함하는 반응 용액을 반응시키면서 일정한 간격으로 형광을 측정함으로써 이루어진다. 이 방법을 통해 형광의 세기가 효소의 양에 비례하는 속도로 감소하는 것을 관찰할 수 있었으며 시간에 따른 형광 변화 그래프로부터 계산한 반응 속도가 효소의 양에 선형으로 비례한다는 것을 확인할 수 있었다. 또 한 가지 중요한 사실은 assay를 하는데 있어서 pH 6.0-8.0의 범위에서 다른 pH 조건을 사용할 수 있었다는 점이다.

• Journal of Microbiology and Biotechnology
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• 제13권6호
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• pp.892-896
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• 2003

High-level expression of the lipase B26 gene from Bacillus pumilus was achieved using Bacillus subtilis Chungkookjang isolated from the Korean traditional fermented bean paste, Chungkookjang. For the secretory production of recombinant lipase B26 in a Bacillus host system, pLipB26 was constructed by ligating the lipase B26 gene into the recently designed Escherichia coli-Bacillus shuttle vector, pLipSM, and that was then transformed into B. subtilis Chungkookjang. Among the various vector, medium, and host combinations, B. subtilis Chungkookjang harboring the pLipB26 exhibited the highest lipase activity in PY medium, and B. subtilis Chungkookjang secreted two times more enzymes than B. subtilis DB 104 under the same condition. When B. subtilis Chungkookjang harboring the pLipB26 was cultured in a 5-1 jar-fermentor containing 21 of a PY medium, the maximum lipase activity (140 U/ml) and production yield (0.68 g/l) were obtained during the late exponential phase from a cell-free culture broth. Although B. subtilis Chungkookjang also secreted extracellular proteases at the late exponential phase, these results suggested the potential of B. subtilis Chungkookjang as a host for the secretory production of foreign proteins.

• 대한본초학회지
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• 제32권4호
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• pp.9-15
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• 2017

Objectives : In this study, the inhibitory activities of Korean Medicine against pancreatic lipase in vitro and biochemical analyses in vivo were measured to determine its possibility as a well-matched sauce material with chicken. Methods : The inhibitory activity on pancreatic lipase enzyme of 11 samples were evaluated in vitro and then 5 samples were selected. The activity of pancreatic lipase was investigated using orlistat as a positive control. Animals were divided into eight groups (n=7). The experimental groups except for normal group were fed 60% high-fat diet for 7 days. 5 samples were orally administered at a dose of 200 mg/kg body weight and orlistat were orally administrated at a dose of 60 mg/kg body weight for 7 days. Biochemical anaylses of 5 samples were executed based on lipid parameters analysis. Results : Korean Medicines with an $IC_{50}$ of below 1 mg/kg were Scutellariae Radix, Gardeniae Fructus, Theae Folium Coptidis Radix, and Mori Cortex Radicis. Body weight change of Mori Cortex Radicis reduced significantly, however fecal triglyceride couldn't regulate effectively. The most excellent inhibitory effect of pancreatic lipase showed in Scutellariae Radix treatment and also regulated significantly serum triglyceride and total cholesterol. Moreover, the supplementation of Coptidis Radix excreted meaningfully triglyceride to fece. Conclusions : In conclusion, Coptidis Radix may exert anti-obesity effect by directly inhibiting pancreatic lipase, which would prevent the absorption of lipid from the small intestine. Besides, Mori Cortex Radicis may led to the decrease of the body weight via the different pathway.

• 한국식품영양과학회지
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• 제24권3호
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• pp.355-361
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• 1995

Lipoprotein lipase(LPL) is an acylglycerol hydrolase and is the extrahepatic enzyme responsible for the hydrolysis of triglyceride-rich plasma lipoproteins. LPL has been isolated from bovine milk by affinity chromatography on heparin-sepharose in 2M NaCl, 5mM barbital buffer, pH 7.4. Para-nitrophenyl butyrate(PNPB) was used as a substrate for the determination of LPL activity. Molecular weight of LPL was 55KD on 10% SDS-PAGE. When the effects of heparin on LPL activation were compared, LPL activity of heparin added group increased approximately 5 times higher than that of heparin non-added groups. These results indicated that heparin involved in the stabilization of LPL structure that led to increase enzyme activity. Furthermore, LPL activity increased about 4 times compared to the absence of heparin at various pH. LPL was stabilized when heparin was added either low or high salt concentrations. With the presence of heparin, NaCl concentration did not affect LPL activity at pH range 6∼9.

• 한국미생물·생명공학회지
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• 제43권4호
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• pp.336-342
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• 2015

선행연구에서 Euptelea pleiosperma가 항산화능과 항염증 활성을 나타내는 유용한 소재임을 처음으로 밝혔다. 본 연구에서는 E. pleiosperma 에탄올 추출물(EPEE)의 항비만 활성을 췌장 리파아제 효소 활성 억제능 및 세포실험모델계를 이용하여 분석하였다. 먼저 EPEE는 농도 의존적으로 lipase 효소 활성을 유의적으로 억제시켰으며, 3T3-L1 preadipocyte에서 지방세포 분화, 세포 내 지방 축적, TG 함량 등을 독성 없이 농도의존적으로 억제하였으며 지방세포 내 중성지방을 유의적으로 분해시키는 것으로 나타났다. 이러한 EPEE의 지방세포 분화 억제능은 핵심 작용 인자인 $C/EBP{\alpha}$, $C/EBP{\beta}$, 그리고 $PPAR{\gamma}$의 유전자 및 단백질 발현조절에서 기인함을 확인하였다. 이러한 결과는 E. pleiosperma가 보유한 췌장 lipase 활성 저해능, 지방세포 분화 억제능, 지방세포 내 지방 분해능을 통한 항비만 활성을 처음으로 밝혀낸 것이며 추후 계속적인 연구를 통해 활성 물질의 규명이 필요할 것으로 판단된다.

• 생약학회지
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• 제41권3호
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• pp.216-220
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• 2010

Obesity is an important risk factor that significantly increases mortality and disease rates in the cardiovascular disease, diabetes, and various diseases. So far, the most powerful way to inhibit fat absorption is pancreatic lipase inhibitors. In this study, we investigated the anti-obesity effect of the extract of Juniperus rigida. Juniperus rigida extract (JRE) had a inhibitory effect on pancreatic lipase activity ($IC_{50}$=8.63 ${\mu}g$/ml). In in vivo oil-emulsion loading test, this extract also inhibited the intestinal fat absorption. In addition, we measured inhibitory effects of JRE on activity of phosphodiesterase (PDE) and hormone sensitive lipase (HSL) among the important enzymes associated with lipolysis. JRE strongly inhibited PDE activity ($IC_{50}$=4.56 ${\mu}g$/ml), whereas inhibitory effect on HSL activity was very weak compared with orlistat. As a result, JRE inhibited the absorption of fat by inhibiting the activity of pancreatic lipase and induced lipolysis through inhibition of PDE activity. Therefore, we suggest that Juniperus rigida may be a potential therapeutic agent improving obesity.

• 한국식품과학회지
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• 제17권2호
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• pp.75-80
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• 1985

Lipase를 유지분해에 이용하기 위한 기초연구로서 Candida cylindracea에서 추출된 lipase를 흡착법에 의해 고정화하고 그의 반응특이성을 본 결과는 다음과 같다. Lipase 흡착에 적합한 흡착제로는 silica gel이 선정되었으며, Silica gel 1.6g에 lipase 47.5 units를 $5^{\circ}C$, PH 7.0에서 100분간 흡착시키는 것이 좋았다. Silica gel에 고정화 시킨 lipase를 유지방과 olive oil의 분해에 적합한 최적온도 및 최적pH는 가용성효소와 비교시 $37^{\circ}C$, pH 7.0으로 변하지는 않았으나 활성의 범위는 넓어졌다. 또한 열안정성 및 pH안정성도 가용성 효소에 비하여 활성의 범위가 넓어졌다. 유지의 분해에 적합한 고정화 효소의 최적효소농도는 유지방의 경우 30g이었으며 올리브유의 경우 80g으로 선정하였다. 이 때 최적기질농도는 유지방 및 올리브유 모두 20%였다. 반응시간에 따른 반응률은 유지방을 이용하여 조사한 결과 가용성 효소는 반응 4시간까지는 급격한 분해를 나타냈으나 고정화 효소는 8시간 까지 급격한 증가를 나타내고 그 이상은 거의 일정하였다. 또한 유리되는 지방산의 profile은 가용성 효소와 비교시 capric acid의 생성은 모두 높았으며, myristic acid의 함량은 높고 butyic acid의 함량은 적었다.