• Journal of Plant Biotechnology
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• 제6권1호
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• pp.63-67
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• 2004

A simple procedure for the extraction of the lipolytic enzyme from rice bran has been developed. High activity of lipolytic enzyme was obtained by first defatting the rice bran to remove lipid components with various extraction conditions. Then, after rove cycles of aqueous extraction, rice bran lipolytic enzyme was purified using micro- and ultrafiltration apparatus. Lipolytic enzyme activity was estimated by its hydrolytic action of tributyrin. The result indicated that the standard activity curve of butyric acid showed that the potential rice bran enzyme is a hydrolytic lipase enzyme. In addition, it showed higher lipolytic activity and specific enzyme activity with further purification by micro- and ultrafiltration. The size of rice bran lipase enzyme was identified through 15 ％ SDS-PAGE. The molecular weight of the rice bran lipase enzyme was 41 kDa.

• 한국산학기술학회:학술대회논문집
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• 한국산학기술학회 2004년도 추계학술대회
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• pp.299-301
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• 2004

A simple procedure for the extraction of the lipolytic enzyme from rice bran has been developed. High activity of lipolytic enzyme was obtained by first defatting the rice bran to remove lipid components with various extraction conditions. Then, after five cycles of aqueous extraction, rice bran lipolytic enzyme was purified using micro- and ultrafiltration apparatus. Lipolytic enzyme activity was estimated by its hydrolytic action of tributyrin. The result indicated that the standard activity curve of butyric acid showed that the potential rice bran enzyme is a hydrolytic lipase enzyme. In addition, it showed higher lipolytic activity and specific enzyme activity with further purification by micro- and ultrafiltration.

• 생명과학회지
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• 제16권4호
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• pp.555-560
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• 2006

유용효소를 생산하는 균주를 가정하수로부터 분리하기 위하여 LBM, R2A, M9배지를 이용하여 다수의 균주를 분리하였다. 분리된 균주 중 1% tributyrin이 첨가된 배지에서 생육 활성대의 형성이 우수한 균주 1종을 최종적으로 선별하여 형태학적, 생리학적, 생화학적 특성을 관찰하였다. 16S rDNA 염기서열 분석결과 Acinetobacter Baumannii (99%)로 Acinetobacter 속에 속하는 균주임을 확인하고, Acinetobacter sp. BD5로 명명하였다. Acinetobacter sp. BD5는 $37^{\circ}C$와 $50^{\circ}C$에서 생육하는 것으로 보아 호열성 균주이며, 1% tributyrin과 oilve oil 첨가된 EL과 CE 고체배지와 Tween 20이 첨가된 LB 고체배지에서 생육활성대의 형성을 확인하여 이 균주가 lipolytic 효소를 생산하는 것으로 나타났다. 효소 활성의 최적 배양조건을 검토하고자 배양시간별 효소 활성을 측정한 결과, 배양 6시간 때인 대수 증식기에 가장 높은 효소활성을 나타내어 비교적 빠른 시간 내에 lipolytic 효소를 생성하는 것으로 나타났다. 또한 효소활성의 최적 온도는 $60^{\circ}C$로 $70-80^{\circ}C$에서 70%이상의 잔존활성을 보이는 것으로 나타나 Acinetobacter sp. BD5는 호열성 균주로 이 균주가 생산하는 lipolytic 효소도 내열성을 보이는 것으로 생각된다. 최적 pH는 9.0이며, pH 9.8-10.6범위에서 50% 이상의 활성이 유지되어 alkaline lipolytic 효소인 것으로 생각되어진다. Acinetobacter sp. BD5가 생산하는 lipolytic 효소는 비교적 넓은 pH 범위와 고온에서 활성이 유지되는 것으로 보아 폐유분해, 세제 합성과 유기물질 합성 등 생물공학분야와 산업적으로 잠재적 가치가 있을 것으로 생각된다.

• Journal of Microbiology and Biotechnology
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• 제17권10호
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• pp.1655-1660
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• 2007

A metagenome is a unique resource to search for novel microbial enzymes from the unculturable microorganisms in soil. A forest soil metagenomic library using a fosmid and soil microbial DNA from Gwangneung forest, Korea, was constructed in Escherichia coli and screened to select lipolytic genes. A total of seven unique lipolytic clones were selected by screening of the 31,000-member forest soil metagenome library based on tributyrin hydrolysis. The ORFs for lipolytic activity were subcloned in a high copy number plasmid by screening the secondary shortgun libraries from the seven clones. Since the lipolytic enzymes were well secreted in E. coli into the culture broth, the lipolytic activity of the subclones was confirmed by the hydrolysis of p-nitrophenyl butyrate using culture supernatant. Deduced amino acid sequence analysis of the identified ORFs for lipolytic activity revealed that 4 genes encode hormone-sensitive lipase (HSL) in lipase family IV. Phylogenetic analysis indicated that 4 proteins were clustered with HSL in the database and other metagenomic HSLs. The other 2 genes and 1 gene encode non-heme peroxidase-like enzymes of lipase family V and a GDSL family esterase/lipase in family II, respectively. The gene for the GDSL enzyme is the first description of the enzyme from metagenomic screening.

• 한국식품영양학회지
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• 제5권1호
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• pp.7-12
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• 1992

This study was devised to observe the inhibitory effect of 7 kinds PG(PG1, PG2, PG3, PG4, PG5, PG6 and PG7) and of 5 kinds PG4(PG41, PG42, PG43, PG44 and PG45) of the acidic polysaccharide fraction from Korean red ginseng on a lipolytic action of Toxohormone-L. Toxohormone-L is a lipolytic factor, found in ascites fluid. of sarcoma-180 bearing mice and of patients with hepatoma. A substance that inhibited the lipolytic action of toxohormone-L was isolated from red ginseng powder. This substance was an acidic polysaccharides In vitro test showed that the inhibitory effect of PG4 and PG43 fraction of the lipolysis by Toxohormone-L was highest percent among other treatments at concentration of 50, 100, 200, 500 and 1,000ug/ml of reaction mixture. And total inhibitory activity(units) of PG1 and PG4, and PG4 s was highest among other treatments at the same concentration.

• Preventive Nutrition and Food Science
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• 제9권1호
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• pp.34-38
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• 2004

High-pungency red pepper and capsaicin modulate circulating hormone levels and induce lipolysis in adipose tissue in vivo. This study was designed to investigate the lipolytic activity of adipocytes by high-pungency red pepper extract in vitro. High-pungency red pepper (var. Chungyang) powder showed 126.1 mg% of capsaicinoid which was 3 x higher than low-pungency red pepper powder (var. Daemyung). To study the effects of high-pungency red pepper extract on lipolytic activity, preadipocytes were separated from the epidermal fat of 14 day-old rats, induced to differentiate into adipocytes and were treated with red pepper extracts. The amount of glycerol released from adipocytes into the culture medium was analysed to measure lipolytic activity. Glycerol release from adipocytes was increased in a dose-dependent manner with high-pungency red pepper extract treatment. However, there was no significant change in the glycerol release when adipocytes were treated with low-pungency red pepper extract. To investigate whether lipolysis by high-pungency red pepper extract is caused by capsaicin, glycerol release was detected after the treatment of adipocytes with capsaicin. Glycerol release was significantly increased by capsaicin. These results suggest that high-pungency red pepper extract might have a direct lipolytic activity in adipocytes that is mediated by capsaicin.

• 생명과학회지
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• 제23권1호
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• pp.110-115
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• 2013

Extracellular 지질분해효소를 생산하는 균주 AR1은 일본 벳부 온천수에서 분리하였다. 분리된 균주의 16s rRNA 염기서열을 분석하고 계통학적으로 분류한 결과, AR1 균주는 신규 Geobacillus sp.에 속하는 것으로 동정되었다. 본 연구는 Geobacillus sp. AR1 균주의 extracellular 지질분해효소 생산을 향상시키기 위한 새로운 방법에 초점을 맞추었다. AR1 균주는 $35{\sim}75^{\circ}C$의 넓은 온도 범위에서 생육하였고 최적온도는 $65^{\circ}C$이었다. 생육을 위한 최적 pH는 6.5인 반면, 효소 생산을 위한 pH는 8.5로 차이점을 보였다. 배양 중에 지질 화합물의 첨가는 지질분해효소 생산을 유도하였고, soybean oil을 대수증식기에 첨가 했을 때 가장 효율적인 유도 효과를 나타내었다. 한편, 계면활성제는 지질분해효소의 생산을 유도하고 세포 내외의 위치에 영향을 줄 수 있다. AR1 균주는 정지기에 Tween 20을 첨가할 경우, 효소의 세포 외 분비 효율이 크게 증가하였다. 이들 결과를 바탕으로 soybean oil과 Tween 20을 각각 대수증식기와 정지기에 첨가함에 따라 extracellular 효소 생산이 대조구에 비해 2.4배 증가하는 것으로 확인 되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제11권4호
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• pp.428-433
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• 1998

The effects of chromium picolinate supplementation in pig diet were evaluated by measuring the in vitro lipogenic and lipolytic activities in adipose tissue and the protein synthetic activity in liver acinar cell in culture. Thirty-two male and thirty-two female pigs were randomly assigned to one of four dietary groups: Control, 100 ppb, 200 ppb, and 400 ppb of Cr in the form of picolinate. The chromium picolinate supplementation (p < 0.01) increased the in vitro lipolytic activity in adipose tissue of pig, but had no effects on lipogenesis. The chromium picolinate effect was greater in female pigs than in male pigs on lipolytic activity. The results from the studies with the liver acinar cells in culture indicated that chromium picolinate supplementation increased protein synthetic activity (p < 0.05). It was observed through this experiment that chromium picolinate functions not only on fat degradation but also on retained protein synthesis.

• Journal of Microbiology and Biotechnology
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• 제19권2호
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• pp.187-193
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• 2009

A metagenomic library of surface-water microbes from the Yangtze River in China was constructed, and a novel esterase, designated as EstY, was isolated and characterized. EstY had 423 amino acids with an estimated molecular mass of 44 kDa and pI of 7.28. It hydrolyzed various p-nitrophenyl esters(acetate, butyrate, caprate, caprylate, laurate, myristate, and palmitate) and its best substrate was p-nitrophenyl caprate(C8). The optimum pH for EstY activity was 9.0 and the optimum temperature was $50^{\circ}C$. Metal ions, such as $Mn^{2+},\;Co^{2+},\;Hg^{2+},\;Zn^{2+},\;and\;Fe^{3+}$, strongly inhibited the activity of EstY, whereas $Mg^{2+}$ was required for maximal activity. Activity remained in the presence of 10% alcohol, acetone, isopropanol, and dimethyl sulfoxide, respectively. An analysis of the amino acid sequence deduced from estY revealed that it had 7 closely related lipolytic enzymes. Moreover, a sequence analysis showed that EstY, like its 7 relatives, did not belong to any known lipolytic enzyme family.

• 한국식품영양학회지
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• 제4권2호
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• pp.149-154
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• 1991

This study was devised to observe the inhibitory effect of 7 kinds of the acidic polysaccharide fraction(PGI, PGa, PG3, PGa, PGs, PG6 and PG7) from Korean white ginseng on a lipolytic action of Toxohormone-L. Toxohormone-L is a lipolytic factor, found in ascites fluid of .sarcoma -180 bearing mice and of patients with hepatoma. A substace that inhibited the lipolytic action of Toxohormone-L was isolated from white ginseng powder. This substance was an acidic polysaccharides. In vitro test showed that the inhibitory effect of PGs fraction of the lipolysis by Toxohormone-L was highest percent among other treatments at concentration of 50, 10, 200, 500 and 1,000 Ug/ml of reaction mixture. And total inhibitory activity(units) of PGI and PG4 was highest among other treatments at the same concentration and that of 10 Ug/ml.