• 산업식품공학
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• 제22권4호
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• pp.295-303
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• 2018

Polyphenol profiles, physicochemical properties, antioxidant activities, and inhibitory effect of adipocyte differentiation of Houttuynia cordata fermented with Lactobacillus brevis B84 were evaluated. Six polyphenols were characterized for this plant by using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), and the results were compared with total phenolic content by a spectrophotometric method. The total amount of the identified polyphenols was lower than that determined by the spectrophotometric method. However, the fermentation process influenced polyphenol composition such as content of vanillic acid and caffeic acid. The phytochemical profiles were evaluated by high-performance liquid chromatography with UV and electrospray ionization mass spectrometry detection ($HPLC-DAD-ESI-MS^n$). Total sugar and reducing sugar contents decreased after fermentation. Antioxidant activities such as DPPH, ABTS, and superoxide anion radical scavenging and reducing power were evaluated to compare the beneficial effect after fermentation. Fermented H. cordata increased the lipolytic effect in 3T3-L1 adipocytes. Overall, the results indicate that the fermentation of H. cordata with L. brevis B84 produces changes of phenolic compounds, antioxidant activity, and lipolytic effect.

• 한국미생물·생명공학회지
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• 제40권2호
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• pp.98-103
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• 2012

경기도 일대에서 수집한 메주 시료에서 지질분해 활성을 나타내는 균주 Y124를 분리하여 동정한 결과 Yarrowia lipolytica와 100% 상동성을 보였다. 분리 균주가 생산하는 lipase의 조효소에 대한 일반적인 특성을 조사한 결과, 탄소원으로 olive oil을 단독으로 사용한 YPO 배지에서 8시간 배양하였을 때 lipase 활성이 가장 높게 나타났다. YPD 배지에서는 lipase 활성이 거의 없었으며, olive oil과 glucose를 모두 포함하는 YPDO 배지에서는 lipase 활성이 YPO 배지 보다 낮았다. 그리고 olive oil 농도에 따른 lipase 활성을 측정한 결과, olive oil 무첨가보다 0.7% 첨가하여 8시간 배양했을 때 lipase 활성이134 U/mL으로 가장 높게 나타나 lipase의 생산이 olive oil의 첨가에 의해 유도되는 것으로 생각된다. 생육온도에 따른 lipase 활성 측정한 결과, $30^{\circ}C$에 배양하였을 때 배양 8시간에 가장 높은 활성이 나타났고, $25^{\circ}C$와 $37^{\circ}C$에 배양하였을 때는 배양 12시간에 활성이 가장 높게 나타났으며, Y124균주의 lipase 활성 최적 온도는 $30^{\circ}C$로 나타났다. 그리고 lipase의 기질 친화도를 확인한 결과 Y124균주가 생산하는 lipase의 경우 p-nitrophenyl octanoate ($C_8$)에서 가장 높은 활성이 나타났다.

• Biomolecules & Therapeutics
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• 제25권3호
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• pp.329-336
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• 2017

Adipogenesis in murine preadipocyte 3T3L-1 has been used as a model system to study anti-obese bioactive molecules. During adipogenesis in 3T3-L1 preadipocytes, we found that capsanthin inhibited adipogenesis ($IC_{50}$; $2.5{\mu}M$) and also showed lipolytic activity in differentiated adipocytes from the preadipocytes ($ED_{50}$; 872 nM). We identified that the pharmacological activity of capsanthin on adipogenesis in 3T3-L1 was mainly due to its adrenoceptor-${\beta}_2$-agonistic activity. In high-fat diet animal model study, capsanthin significantly enhanced spontaneous locomotive activities together with progressive weight-loss. The capsanthin-induced activation of kinetic behavior in mice was associated with the excessive production of ATP initiated by both the enhanced lipolytic activity together with accelerated oxidation of fatty acids due to the adrenoceptor ${\beta}_2$-agonistic activity of capsanthin. Capsanthin also dose-dependently increased adiponectin and p-AMPK activity in high fat diet animals, suggesting that capsanthin has both anti-obesity and insulin sensitizing activities.

• Asian-Australasian Journal of Animal Sciences
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• 제33권10호
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• pp.1590-1598
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• 2020

Objective: The objective of this study was to evaluate the effects of lysophospholipids (LPL) supplementation on rumen fermentation, degradability, and microbial diversity in forage with high oil diet in an in vitro system. Methods: Four experimental treatments were used: i) annual ryegrass (CON), ii) 93% annual ryegrass +7% corn oil on a dry matter (DM) basis (OiL), iii) OiL with a low level (0.08% of dietary DM) of LPL (LLPL), and iv) OiL with a high level (0.16% of dietary DM) of LPL (HLPL). An in vitro fermentation experiment was performed using strained rumen fluid for 48 h incubations. In vitro DM degradability (IVDMD), in vitro neutral detergent fiber degradability, pH, ammonia nitrogen (NH₃-N), volatile fatty acid (VFA), and microbial diversity were estimated. Results: There was no significant change in IVDMD, pH, NH₃-N, and total VFA production among treatments. The LPL supplementation significantly increased the proportion of butyrate and valerate (Linear effect [Lin], p = 0.004 and <0.001, respectively). The LPL supplementation tended to increase the total bacteria in a linear manner (p = 0.089). There were significant decreases in the relative proportions of cellulolytic (Fibrobacter succinogenes and Ruminococcus albus) and lipolytic (Anaerovibrio lipolytica and Butyrivibrio proteoclasticus) bacteria with increasing levels of LPL supplementation (Lin, p = 0.028, 0.006, 0.003, and 0.003, respectively). Conclusion: The LPL supplementation had antimicrobial effects on several cellulolytic and lipolytic bacteria, with no significant difference in nutrient degradability (DM and neutral detergent fiber) and general bacterial counts, suggesting that LPL supplementation might increase the enzymatic activity of rumen bacteria. Therefore, LPL supplementation may be more effective as an antimicrobial agent rather than as an emulsifier in the rumen.

• The Korean Journal of Physiology and Pharmacology
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• 제25권4호
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• pp.333-339
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• 2021

Injection lipolysis or mesotherapy gained popularity for local fat dissolve as an alternative to surgical liposuction. Phosphatidylcholine (PPC) and aminophyl-line (AMPL) are commonly used compounds for mesotherapy, but their efficacy and safety as lipolytic agents have been controversial. Glycerophosphocholine (GPC) is a choline precursor structurally similar to PPC, and thus introduced in aesthetics as an alternative for PPC. This study aimed to evaluate the effects of GPC on adipocytes differentiation and lipolysis and compared those effects with PPC and AMPL using in vitro and in vivo models. Adipogenesis in 3T3-L1 was measured by Oil Red O staining. Lipolysis was assessed by measuring the amount of glycerol released in the culture media. To evaluate the lipolytic activity of GPC on a physiological condition, GPC was subcutaneously injected to one side of inguinal fat pads for 3 days. Lipolytic activity of GPC was assessed by hematoxylin and eosin staining in adipose tissue. GPC significantly suppressed adipocyte differentiation of 3T3-L1 in a concentration-dependent manner (22.3% inhibition at 4 mM of GPC compared to control). Moreover, when lipolysis was assessed by glycerol release in 3T3-L1 adipocytes, 6 mM of GPC stimulated glycerol release by two-fold over control. Subcutaneous injection of GPC into the inguinal fat pad of mice significantly reduced the mass of fat pad and the size of adipocytes of injected site, and these effects of GPC were more prominent over PPC and AMPL. Taken together, these results suggest that GPC is the potential therapeutic agent as a local fat reducer.

• Journal of Ginseng Research
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• 제15권2호
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• pp.106-111
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• 1991

This study was devised to obserL'e the inhibitory effects of 7 kinds of ginsenosides on a lipolytic action of Toxohormone-L. The ginsenosides used in this experiment were -Rbl, -Rbl, - Rc, - Re, - Rgl, and - Rg2 prepared from Korean red ginseng. Toxohormene-L was partially purified by centrifigation from the ascites fluids of Sarcoma-180 bearing mice. In vitro test showed that the inhibitory effect of - Rb2 on the lipolysis by Toxohormone-L was highest percent among other treatments at concentration of 100 $\mu\textrm{g}$/ml and 500 $\mu\textrm{g}$/ml of reaction mixture. And total inhibitory activity (units) of - Rb2 was also highest among other treatments at the same concentration. However, in vivo test, body weight gain of Sarcoma-180 bearing mice decreased significantly by administration of - Rg2 compared to those of the control or other ginsenosides treated groups.

• 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.

• Journal of Microbiology and Biotechnology
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• 제17권4호
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• pp.604-610
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• 2007

A psychrotrophic strain 7195 showing extracellular lipolytic activity towards tributyrin was isolated from deep-sea sediment of Prydz Bay and identified as a Psychrobacter species. By screening a genomic DNA library of Psychrobacter sp. 7195, an open reading frame of 954 bp coding for a lipase gene, lipA1, was identified, cloned, and sequenced. The deduced LipA1 consisted of 317 amino acids with a molecular mass of 35,210 kDa. It had one consensus motif, G-N-S-M-G (GXSXG), containing the putative active-site serine, which was conserved in other cold-adapted lipolytic enzymes. The recombinant LipA1 was purified by column chromatography with DEAE Sepharose CL-4B, and Sephadex G-75, and preparative polyacrylamide gel electrophoresis, in sequence. The purified enzyme showed highest activity at $30^{\circ}C$, and was unstable at temperatures higher than $30^{\circ}C$, indicating that it was a typical cold-adapted enzyme. The optimal pH for activity was 9.0, and the enzyme was stable between pH 7.0-10.0 after 24h incubation at $4^{\circ}C$. The addition of $Ca^{2+}\;and\;Mg^{2+}$ enhanced the enzyme activity of LipA1, whereas the $Cd^{2+},\;Zn^{2+},\;CO^{2+},\;Fe^{3+},\;Hg^{2+},\;Fe^{2+},\;Rb^{2+}$, and EDTA strongly inhibited the activity. The LipA1 was activated by various detergents, such as Triton X-100, Tween 80, Tween 40, Span 60, Span 40, CHAPS, and SDS, and showed better resistance towards them. Substrate specificity analysis showed that there was a preference for trimyristin and p-nitrophenyl myristate $(C_{14}\;acyl\; groups)$.

• Journal of Microbiology and Biotechnology
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• 제31권3호
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• pp.483-491
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• 2021

Two putative genes, lip29 and est29, encoding lipolytic enzymes from the thermophilic bacterium Geobacillus thermocatenulatus KCTC 3921 were cloned and overexpressed in Escherichia coli. The recombinant Lip29 and Est29 were purified 67.3-fold to homogeneity with specific activity of 2.27 U/mg and recovery of 5.8% and 14.4-fold with specific activity of 0.92 U/mg and recovery of 1.3%, respectively. The molecular mass of each purified enzyme was estimated to be 29 kDa by SDS-PAGE. The alignment analysis of amino acid sequences revealed that both enzymes belonged to GDSL lipase/esterase family including conserved blocks with SGNH catalytic residues which was mainly identified in plants before. While Est29 showed high specificity toward short-chain fatty acids (C4-C8), Lip29 showed strong lipolytic activity to long-chain fatty acids (C12-C16). The optimal activity of Lip29 toward p-nitrophenyl palmitate as a substrate was observed at 50℃ and pH 9.5, respectively, and its activity was maintained more than 24 h at optimal temperatures, indicating that Lip29 was thermostable. Lip29 exhibited high tolerance against detergents and metal ions. The homology modeling and substrate docking revealed that the long-chain substrates showed the greatest binding affinity toward enzyme. Based on the biochemical and insilico analyses, we present for the first time a GDSL-type lipase in the thermophilic bacteria group.

• Journal of Nutrition and Health
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• 제26권2호
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• pp.119-130
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• 1993

본 시험은 Sprague-Dawley 계통 흰쥐 수컷에게 지방의 종류와 식이급여 횟수를 달리하면서 급여하였을 때 성장 및 지방대사에 미치는 영향을 알아보고자 4종류의 지방(팜유, 우지, 대두유 및 경화대두유) 및 2종류의 식이급여형태(ad-libitum, 4$\times$meal feeding)를 달리하여 4$\times$2 요인시험으로 실시하였다. 4주간 시험식이를 ad-libitum(AL)군은 식이를 제한없이 자유롭게 섭취하도록 하였으며, meal feeding(MF)군은 1일 1회 즉, 09:00~12:00까지만 식이를 제한없이 급여하였다. 사양시험 종료후 간조직에서의 콜레스테롤합성 능력을 알아보기 위해 흰쥐의 간조직을 채취하여 in vitro 배양 시험을 실시하였다. 또한 간과 지방조직에서의 지방합성 및 분해 능력을 알아보기 위해 사양시험이 끝난 흰쥐의 간과 지방조직을 채위하여 in vitro 세포배양에서의 지방합성 능력과 지방분해 능력을 측정하였다. 본 시험에서 얻어진 결과를 요약하면 다음과 같다. 1) 성장하는 흰쥐의 증체량, 식이섭취량 및 식이효율은 AS(ad-libitum+hydrogenated soybean oil)군이 다른군에 비해 유의하게(p<0.05) 증가하였다. 식이지방의 종류에 의한 영향은 받지 않고, 식이급여형태에 의한 차이를 보여 MF군이 AL군에 비해 식이섭취량이 감소하였고, 중체량도 60%정도로 유의성있게(p<0.05)감소했다. 2) 간조직의 콜레스테롤합성 능력은 식이급여형태에 의한 차이를 보여 MF군이 AL군에 비해 간조직의 콜레스테롤합성 능력이 증가되는 경향을 (p<0.001) 보였는데 특히 불포화도가 높은 대두유군에서 유의성있게(p<0.05) 증가되었다. 3) 간조직의 in vitro 지방합성 능력은 MP(meal feeding+palm oil)군이 AP(ad-libitum+palm oil)군보다, MHS(meal feeding+hydrogenated soybean oil)군이 AHS(ad-libitum+hydrogenated soybean oil)군보다 유의하게(p<0.05) 증가했고, AB(ad-libium+beef tallow)군과 MB(meal feeding+beef tallow)군에서 지방합성 능력이 현저하게 증가되는 경향을(p<0.05)보였다. 식이급여형태나 식이지방의 종류에 대한 차이는 보이지 않았다. 간조직에서의 in virto 지방분해 능력은 MF군이 AL군에 비해 유의하게(P<0.001) 증가했고, 식이지방의 종류에 의한 차이는 나타나지 않았다. 4) 지방조의 in vitro 시험에 의하면 MF식이가 AL식이에 비해 지방합성 능력이 유의성있게(p<0.05) 증가하였으나, 지방분해 능력은 저하되었다(p<0.001). 식이지방의 종류에 의한 영향을 받아 우지군이 다른 지방군에 비해 지방합성 능력이 유의성있게(p<0.001) 증가되는 경향을 보였으나, 지방분해 능력은 식이지방의 종류에 의한 영향을 받지 않았다.