• Title/Summary/Keyword: lipolytic activity

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Purification and Biochemical Analysis of Rice Bran Lipase Enzyme

  • Kim, Young Hee
    • Journal of Plant Biotechnology
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    • v.6 no.1
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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.

Purification and Biochemical Analysis of Rice Bran Lipase Enzyme (쌀겨로부터 lipase 효소의 정제 및 생화학적인 분석)

  • Kim Younghee
    • Proceedings of the KAIS Fall Conference
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    • 2004.11a
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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.

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Isolation and Characterization of Acinetobacter sp. BD5 Producing Lipolytic Enzyme (Lipolytic 효소를 생산하는 Acinetobacter sp. BD5 균주의 분리 및 특성)

  • Park, In-Hye;Kim, Sun-Hee;Lee, Sang-Cheol;Ahn, Soon-Cheol;Kim, Cheol-Min;Choi, Yong-Lark
    • Journal of Life Science
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    • v.16 no.4
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    • pp.555-560
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    • 2006
  • A bacterium producing novel lipolytic enzyme was isolated from house sewage and identified as Acinetobacter sp. BD5 based on physiological characterization and 16S rDNA sequencing. The lipolytic activity of Acinetobacter sp. BD5 was tested using an EL agar medium and CE agar medium supplemented with 1% tributyrin and olive oil, respectively. The formation of a clear zone around the colony was detected by agar medium supplemented with 1% tributyrin and olive oil, respectively and Acinetobacter sp. BD5 formed powder-like zone around the colony on LB agar medium containing Tween 20. The quantitative lipolytic activity was determined by using p-NP butyrate as substrate. Acinetobacter sp. BD5 secreted the lipolytic enzyme during exponential growth phase, reaching a maximum amount after 6 hours of incubation. The lipolytic enzyme was found to be optimally active at $60^{\circ}C$ and retained more than 70% at $70-80^{\circ}C$. It displayed a high degree of activity in a pH of 7.0 to 10.6, with an optimal pH of 9.0.

Selection and Characterization of Forest Soil Metagenome Genes Encoding Lipolytic Enzymes

  • Hong, Kyung-Sik;Lim, He-Kyoung;Chung, Eu-Jin;Park, Eun-Jin;Lee, Myung-Hwan;Kim, Jin-Cheol;Cho, Gyung-Ja;Cho, Kwang-Yun;Lee, Seon-Woo
    • Journal of Microbiology and Biotechnology
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    • v.17 no.10
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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.

Inhibitory Activity of Acidic Polysaccharides of Korean Red Ginseng on Lipolytic Action of Toxohormone-L from Cancerous Ascites Fluid (홍삼 산성 다당체 성분의 암독소 호르몬-L에 의한 지방 분해 저해 활성)

  • 황윤경;이성동
    • The Korean Journal of Food And Nutrition
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    • v.5 no.1
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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.

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Increased Lipolytic Activity by High-pungency Red Pepper Extract(var. Chungyang) in Rat Adipocytes in vitro

  • Do, Myoung-Sool;Hong, Sung-Eui;Ha, Jung-Heun;Park, Sun-Mi;Ahn, In-Sook;Yoon, Ji-Young;Park, Kun-Young
    • Preventive Nutrition and Food Science
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    • v.9 no.1
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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.

Culture Conditions for Improving Extracellular Lipolytic Enzyme Production by a Novel Thermophilic Geobacillus sp. AR1 (신규 고온성 Geobacillus sp. AR1의 extracellular 지질분해효소 생산을 위한 배양조건)

  • Park, Su-Jin;Jeon, Sung-Jong
    • Journal of Life Science
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    • v.23 no.1
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    • pp.110-115
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    • 2013
  • A microorganism (strain AR1) producing an extracellular lipolytic enzyme was isolated from hot springs located in Beppu, Japan. Phylogenetic analysis based on the 16S rDNA sequence and biochemical studies indicated that AR1 belongs to the genus Geobacillus. This study focused on novel strategies to increase extracellular lipolytic enzyme production by this novel Geobacillus sp. AR1. Cultures of the AR1 strain grew within a wide temperature range (from 35 to $75^{\circ}C$); the optimum temperature was $65^{\circ}C$. The pH for optimal growth was 6.5, whereas the optimum pH for lipolytic enzyme production was 8.5. The presence of oils in the culture medium led to improvements in lipolytic enzyme activity. Soybean oil was the most efficient inducer, and it yielded better results when added in the exponential phase. On the other hand, the addition of chemical surfactants led to lipolytic enzyme production. Their addition to the culture could affect the location of the enzyme activity. The addition of Tween 20 in the stationary phase significantly increased the proportion of the extracellular enzyme activity. According to the results, following the addition of soybean oil and Tween 20 in the exponential and stationary phases, the extracellular lipolytic activity was increased 2.4-fold compared with that of a control.

Effects of Chromium Picolinate on In Vitro Lipogenesis and Lipolysis in Adipose Tissue and Protein Synthesis in Liver Tissue of Pigs

  • Choi, Y.J.;Kim, H.G.;Cho, J.S.;Chung, I.B.;Kim, Y.H.;Han, I.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.11 no.4
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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.

Identification of Novel Esterase from Metagenomic Library of Yangtze River

  • Wu, Chao;Sun, Baolin
    • Journal of Microbiology and Biotechnology
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    • v.19 no.2
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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.

Effect of Acidic Polysaccharides of Korean White Ginseng on Lipolytic Action of Toxohormone-L from Cancerous Ascites Fluid (암독소 호르몬-L이 유발하는 체지방분해에 고려백삼 산성다당체 성분이 미치는 영향)

  • 이성동
    • The Korean Journal of Food And Nutrition
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    • v.4 no.2
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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.

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