• 제목/요약/키워드: Lipase assay

검색결과 70건 처리시간 0.023초

우렁쉥이 추출물의 항산화 및 리파아제 저해활성 효과 (Antioxidant and Anti-lipase Activity in Halocynthia roretzi Extracts)

  • 권태형;김진기;김태완;이진욱;김준태;서현주;김민정;김충곤;전득산;박년호
    • 한국식품과학회지
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    • 제43권4호
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    • pp.464-468
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    • 2011
  • 본 연구에서는 동해안에 서식하는 우렁쉥이의 식품산업의 소재와 기능성을 알아보기 위하여 우렁쉥이 추출물을 이용하여 이들로부터 활성산소종의 소거활성, 즉 항산화제와 비만 예방의 효능제로서 생리활성을 살펴보고자 하였다. 우렁쉥이의 에탄올, 메탄올, 물 추출물을 이용하여 폴리페놀 및 플라보노이드 함량을 측정한 결과 에탄올 추출물과 메탄올 추출물이 물 추출물보다 약 1.5배 높게 나타났고 플라보노이드 함량 또한 약 8배 이상 높게 나타났다. 항산화 효능을 측정한 결과 DPPH 라디칼 소거능에서는 메탄올 추출물이 1176.99 ${\mu}g/mL$로 가장 높았으며 물 추출물에 비해 약 2배정도 높은 효능을 보였다. ABTS 라디칼 소거능 활성을 측정한 결과 875.07 ${\mu}g/mL$로 메탄올 추출물이 가장 높은 효능을 보였고 DPPH 라디칼 소거능 활성과 유사한 경향을 보였다. 환원력 측정은 메탄올 추출물 31.7%, 에탄올 추출물 32.2% 그리고 물 추출물 26.4%의 순으로 나타났으며 금속 봉쇄력의 경우는 물 추출물이 가장 높은 활성을 보였다. 폴리페놀 함량이 높은 물질이 항산화 효능과의 상관관계가 있음을 알 수 있었고 지방 분해 측정 결과 에탄올 추출물이 6004.76 ${\mu}g/mL$, 메탄올 추출물이 12021.56 ${\mu}g/mL$로 측정되었으며 에탄올 추출물에는 지방 분해 억제 효과가 있는 것으로 생각된다. 결과적으로 우렁쉥이는 항산화 효과와 비만예방 개선과 다양한 식품 소재로의 활용 가능성이 있을 것으로 사료되며, 이러한 결과가 실제로 체내에서 적용되는지는 in vivo 연구를 통하여 살펴볼 필요가 있는 것으로 사료된다.

Bioluminescent assay of Phospholipase C Using A Luminescent Marine Mutant Bacterium Vibrio harveyi M-17

  • Cho, Ki-Woong;Mo, Sang-Jun;Lee, Hyi-Seung;Park, Jung-Rae;Jongheon Shin
    • Journal of Microbiology
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    • 제38권3호
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    • pp.150-155
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    • 2000
  • A bioluminescent assay method for detecting the activity of phospholipase C(PLC; phosphatidyl choline cholinephosphohydrolase, EC 3.1.4.3) was developed using bioluminescent marine bacteria. Phospholipase C from Bacillus cereus and sn-1,2- dimyristoyl glycerol was further hydrolyzed with lipase from Candida ecylidracea. The hydrolyzed myristic acid was quantified using a dark mutant of Vibrio harveyi (designated as M-17). The in vivo light intensity of which was stimulated specifically up to one thousand fold in the presence of myristic acid. The rates of the hdrolysis of the DMPC substrate by the phospholipase measured by the luminescence method were linear with time and the were estalished to detect as little as 0.1 mUnit of phospholipase C and 5 nM of myristic acid production.

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Lectin Activity and Chemical Characteristics of Escherichia coli, Lactobacillus spp. and Bifidobacterium spp. from Gastrointestinal Mucosa of Growing Pigs

  • Gao, W.;Meng, Q.X.
    • Asian-Australasian Journal of Animal Sciences
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    • 제17권6호
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    • pp.863-868
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    • 2004
  • Lectin activities and chemical characteristics of Escherichia coli, Lactobacillus spp. and Bifidobacterium spp. originating from the porcine cecal mucosal layer were studied based on hemagglutination assay (HA) and hemagglutination inhibition assay (HIA). Although all the bacterial strains were able to agglutinate erythrocytes of porcine or rabbit origin, much higher HA titers were consistently observed for Lactobacillus spp. than for E. coli or for Bifidobacterium spp. A remarkable reduction in HA titers occurred by the treatment of E. coli and Lactobacillus spp. with protease or trypsin and of Bifidobacterium spp. with protease, trypsin or periodate. There were no significant effects on the HA titers of the three groups of bacteria after the treatment with lipase. Hemagglutination of E. coli was strongly inhibited by D (+)-mannose and D (+)-galactose; Lactobacillus spp. by $\alpha$-L-rhamnose and methyl-$\beta$-galactopyranoside; Bifidobacterium spp. by D (+)-alactose, $\alpha$-L-rhamnose, $\alpha$-L-fucose, L (+)-arabinose, D (+)-mannose, D (-)-fructose at a relatively low concentration (1.43 to 3.75 mg/ml). These results, combined with the enhanced HA activities of the three bacterial strains by modification of rabbit erythrocytes with neuraminidase and abolished HA activity of E. coli after treatment with $\beta$-galactosidase, indicate that it might be the glycoproteinous substances surrounding the surface of the bacterial cells that are responsible for the adhesions of these microorganisms by recognizing the specific receptors on the red blood cell.

Biosurfactant Production from Novel Air Isolate NITT6L: Screening, Characterization and Optimization of Media

  • Vanavil, B.;Perumalsamy, M.;Rao, A. Seshagiri
    • Journal of Microbiology and Biotechnology
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    • 제23권9호
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    • pp.1229-1243
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    • 2013
  • In this paper, an air isolate (NITT6L) has been screened based on hemolytic activity, emulsification activity, drop collapsing test, and oil displacement test, as well as lipase activity. It was found that strain NITT6L was able to reduce the surface tension of the medium from 61.5 to 39.83 mN/m and could form stable emulsions with tested vegetable oils. Morphological, biochemical, 16S rRNA sequencing analyses, and fatty acid methyl ester analysis using gas chromatography confirmed that the air isolate under study was Pseudomonas aeruginosa. Characterization of the biosurfactant using agar double diffusion assay revealed that the biosurfactant was anionic in nature, and CTAB-methylene blue assay and Molisch test revealed its glycolipid nature. The FT-IR spectrum confirmed that the crude biosurfactant was a rhamnolipid. Using unoptimized medium containing sucrose as the carbon source, the isolate was found to produce 0.3 mg/ml of rhamnolipid in batch cultivation (shake flask) at $37^{\circ}C$ and pH 7. Optimization of the medium components was carried out using design of experiments and the yield of rhamnolipid has been enhanced to 4.6 mg/ml in 72 h of fermentation.

Rapid Detection of Virulence Factors of Aeromonas Isolated from a Trout Farm by Hexaplex-PCR

  • Nam, In-Young;Joh, Ki-Seong
    • Journal of Microbiology
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    • 제45권4호
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    • pp.297-304
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    • 2007
  • The detection of virulence factors of Aeromonas is a key component in determining potential pathogenicity because these factors act multifunctionally and multifactorially. In this study water samples were collected from a trout farm on a seasonal basis, and diseased fish and Aeromonas species were isolated and identified. For rapid detection of six virulence factors of isolated Aeromonas, a hexaplex-polymerase chain reaction (hexaplex-PCR) assay was used. The detected virulence factors include aerolysin (aer), GCAT (gcat), serine protease (ser), nuclease (nuc) lipase (lip) and lateral flagella (laf). The dominant strain found in our isolates was Aeromonas sobria, and the dominant virulence factors were aer and nuc for all seasons. We confirmed that A. sobria and two of the virulence genes (aer and nuc) are related. We proposed a method by which one can identify the major strains of Aeromonas: A. hydrophila, A. sobria, A. caviae, and A. veronii, using hexaplex-PCR.

Antiobesity effects of the water-soluble fraction of the ethanol extract of Smilax china L. leaf in 3T3-L1 adipocytes

  • Kang, Yun Hwan;Kim, Kyoung Kon;Kim, Dae Jung;Choe, Myeon
    • Nutrition Research and Practice
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    • 제9권6호
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    • pp.606-612
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    • 2015
  • BACKGROUND/OBJECTIVES: Several medicinal properties of Smilax china L. have been studied including antioxidant, anti-inflammatory, and anti-cancer effects. However, the antiobesity activity and mechanism by which the water-soluble fraction of this plant mediates its effects are not clear. In the present study, we investigated the lipolytic actions of the water-soluble fraction of Smilax china L. leaf ethanol extract (wsSCLE) in 3T3-L1 adipocytes. MATERIALS/METHODS: The wsSCLE was identified by measuring the total polyphenol and flavonoid content. The wsSCLE was evaluated for its effects on cell viability, lipid accumulation, glycerol, and cyclic adenosine monophosphate (cAMP) contents. In addition, western blot analysis was used to evaluate the effects on protein kinase A (PKA), PKA substrates (PKAs), and hormone-sensitive lipase (HSL). For the lipid accumulation assay, 3T3-L1 adipocytes were treated with different doses of wsSCLE for 9 days starting 2 days post-confluence. In other cell experiments, mature 3T3-L1 adipocytes were treated for 24 h with wsSCLE. RESULTS: Results showed that treatment with wsSCLE at 0.05, 0.1, and 0.25 mg/mL had no effect on cell morphology and viability. Without evidence of toxicity, wsSCLE treatment decreased lipid accumulation compared with the untreated adipocyte controls as shown by the lower absorbance of Oil Red O stain. The wsSCLE significantly induced glycerol release and cAMP production in mature 3T3-L1 cells. Furthermore, protein levels of phosphorylated PKA, PKAs, and HSL significantly increased following wsSCLE treatment. CONCLUSION: These results demonstrate that the potential antiobesity activity of wsSCLE is at least in part due to the stimulation of cAMP-PKA-HSL signaling. In addition, the wsSCLE-stimulated lipolysis induced by the signaling is mediated via activation of the ${\beta}$-adrenergic receptor.

급성 췌장염모델에서 전침의 치료효과 (Therapeutic Effects of Electroacupuncture on Cholecystokinin-octapeptide-induced Acute Pancreatitis Models)

  • 정상수;윤지원;정경아;이종덕;배선준;조장희;성강경
    • Journal of Acupuncture Research
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    • 제22권5호
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    • pp.57-66
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    • 2005
  • Objectives : We examined the effects of electroacupuncture on the cholecystokinin-octapeptide-induced acute pancreatitis in rats. Methods : Rats were administered with $75{\mu}g/kg$ cholecystokinin-octapeptide subcutaneously three times (1, 3 and 5h after shaving) for 5days. Three days after finishing cholecystokinin-octapeptide administration, high frequency electroacupuncture (100Hz) and low frequency electroacupuncture (2Hz) were applied to acupoint equivalent to ST36 (Zusanli) for 7 days. The author determined the pancreatic weight/body weight ratio, the levels of pancreatic heat shock protein HSP60 and HSP72. The author also assay the secretion of ${\beta}-amylase$, lipase and pro-inflammatory cytokines in serum. Repeated cholecysokinin-octapeptide treatment resulted in the typical laboratory and morphological changes of experimentally induced pancreatitis. Results : Eelectroacupuncture significantly decreased the pancreatic weight/body weight ratio in cholecystokinin-octapeptide-induced acute pancreatitis, increased the pancreatic levels of HSP60 and HSP72, and decreased ${\beta}-amylase$ and lipase levels in cholecystokinin-octapeptide-induced acute pancreatitis. Additionally, the secretion of $Interleukin-1{\beta}$ and tumor necrosis $factor-{\alpha}$ was decreased in the animals treated with electroacupuncture. Conclusion : These results suggest that electroacupuncture may have protective effects against cholecystokinin-octapeptide-induced acute pancreatitis.

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생 마늘과 절인 마늘의 In Vitro 항비만 효과 (In Vitro Anti-Obesity Effects of Raw Garlic and Pickled Garlic)

  • 이다빈;표영희
    • 한방비만학회지
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    • 제21권2호
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    • pp.69-79
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    • 2021
  • Objectives: This study aimed to compare the anti-obesity effects of raw and pickled garlic in vitro in 3T3-L1 preadipocytes. Methods: The pickled garlic samples comprised the following: garlic aged in vinegar (VG), garlic aged in soy sauce, and vinegar (1:1, v/v) (PG) and raw garlic (RG) as control. Hexane, butanol, and distilled water were used to prepare the fractions. The pancreatic lipase inhibitory activity was used as a measure of anti-obesity effects of the extracts. The lipid droplet accumulation and triglyceride content in the 3T3-L1 cells were measured using Oil red O staining and triglyceride assay kits, respectively. The adipogenesis related protein expression levels were analyzed using the kits and the western blot method. Results: The pancreatic lipase inhibitory activity of the garlic extracts (VG, PG, RG) was the highest in the butanol fraction, and the inhibitory effect was the highest in RG, followed by PG and VG. All garlic butanol extracts suppressed triglyceride accumulation in differentiated adipocytes (P<0.05) through the activation of cyclic adenosine monophosphate (AMP), AMP-activated protein kinase, carnitine palmitoyl transferase-1, and the inhibition of fatty acid synthase. Raw garlic extracts significantly inhibited the expression of proteins involved in adipogenesis as compared to pickled garlic. Conclusions: Raw garlic has the potential to be an effective natural material for reducing obesity compared to pickled garlic with vinegar or soy sauce.

Cydonia oblonga Miller fruit extract exerts an anti-obesity effect in 3T3-L1 adipocytes by activating the AMPK signaling pathway

  • Hyun Sook Lee;Jae In Jung;Jung Soon Hwang;Myeong Oh Hwang;Eun Ji Kim
    • Nutrition Research and Practice
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    • 제17권6호
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    • pp.1043-1055
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    • 2023
  • BACKGROUND/OBJECTIVES: The fruit of Cydonia oblonga Miller (COM) is used traditionally in Mediterranean region medicine to prevent or treat obesity, but its mechanism of action is still unclear. Beyond a demonstrated anti-obesity effect, the fruit was tested for the mechanism of adipogenesis in 3T3-L1 preadipocytes. MATERIALS/METHODS: 3T3-L1 preadipocytes were cultured for 8 days with COM fruit extract (COME) at different concentrations (0-600 ㎍/mL) with adipocyte differentiation medium. The cell viability was measured using an MTT assay; triglyceride (TG) was stained with Oil Red O. The expression levels of the adipogenesis-related genes and protein expression were analyzed by reverse transcription polymerase chain reaction and Western blotting, respectively. RESULTS: COME inhibited intracellular TG accumulation during adipogenesis. A COME treatment in 3T3-L1 cells induced upregulation of the adenosine monophosphate-activated protein kinase (AMPK)α phosphorylation and downregulation of the adipogenic transcription factors, such as sterol regulatory element-binding protein 1c, peroxisome proliferator-activated receptor γ, and CCAAT/enhancer binding protein α. The COME treatment reduced the mRNA expression of fatty acyl synthetase, adenosine triphosphate-citrate lyase, adipocyte protein 2, and lipoprotein lipase. It increased the mRNA expression of hormone-sensitive lipase and carnitine palmitoyltransferase I in 3T3-L1 cells. CONCLUSIONS: COME inhibits adipogenesis via the AMPK signaling pathways. COME may be used to prevent and treat obesity.

난백(Egg White)에서 추출한 리소좀 추출물(LOE)의 미백 효능 및 피부장벽에 관한 연구 (Study on the Whitening Efficacy and Skin Barrier by Lysosome-related Organelle Extract (LOE) from Egg White)

  • 최다희;전경찬;윤지희;민지호;박시준;김정수;황이택;황형서
    • 대한화장품학회지
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    • 제45권4호
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    • pp.389-397
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    • 2019
  • 리소좀(lysosome)은 진핵세포에서 에너지 대사 및 세포 내 소화 작용에 관여하는 세포 소기관으로 protease, nuclease, glycosidase, lipase, phosphatase 들이 다수 존재한다. 우리는 선행 연구결과들을 통해 난백 리소좀의 멜라닌 색소 탈색능을 보고하였다[8]. 그러나 B16F10 melanocyte 세포주에서 난백 리소좀에 의한 멜라닌 함량 변화 및 피부장벽 조절 연구는 거의 보고되지 않았다. 따라서 우리는 계란 난백(egg white)으로부터 추출한 lysosome-related organelle extract (LOE)에 의한 세포 내 멜라닌 함량 변화 및 피부장벽 강화 효과를 규명하고자 하였다. 먼저 LOE의 미백 효능을 확인하기 위해 B16F10 세포주를 이용하여 세포독성 평가를 진행하였다. B16F10 세포주에서 LOE에 의한 세포독성은 0에서 20 mg/mL 농도에서 관찰되지 않았으나, 40 mg/mL 부터 세포독성이 관찰되어 이후 모든 실험에서 최대 농도값을 20 mg/mL로 설정하였다. 먼저 LOE를 이용한 melanin contents assay 결과, 음성 대조군인 α-MSH 처리군 대비 LOE 처리군 5, 10, 20 mg/mL 농도에서 61.5 ± 4.0%, 61.4 ± 7.3%, 58.3 ± 8.3%로 세포 내 멜라닌 함량이 감소되는 것을 확인하였고, 20 mg/mL 농도 조건에서 MITF 발현 억제도 관찰하였다. LOE의 피부 장벽에 미치는 영향을 관찰하기 위해 각질형성세포주(HaCaT)를 이용하여 TEER (trans-epithelial electrical resistance) assay를 수행한 결과, LOE에 의해 농도 의존적으로 TEER 저항값이 증가하여 LOE가 피부장벽 강화에도 효과가 있음을 알 수 있었다. 또한 피부 염증 유발을 위한 TNF-α 처리조건에서도 LOE는 TEER 저항값을 증가시켜 염증 유발 조건에서도 LOE에 의해 피부장벽이 정상적으로 회복되었음을 알 수 있었다. 마지막으로 cell migration assay를 통해 LOE에 의한 세포이동 촉진 효과를 관찰한 결과, LOE는 세포분열 및 세포이동을 촉진시켰다. 위 결과들을 통해 LOE는 미백 기능 뿐 아니라 피부재생 및 피부장벽 강화에도 효과를 나타내는 소재이며, 효소안정화 및 제형화 기술이 접목된다면 향후 새로운 미백 기능성 화장품 소재로도 개발될 수 있을 것이다.