Lipolytic genes

  • 文章类型: Journal Article
    Virgibacillusspp.作为一种有效的发酵剂,可以加速鱼酱和虾酱的发酵。然而,负责其适应和生物技术潜力的潜在分子机制仍然难以捉摸。因此,本研究的重点是来自越南高盐发酵虾酱的嗜盐细菌dokdonensisT4.6的嗜盐细菌的表型和基因组分析。基因组草案包含4,096,868bp和3780个预测编码序列。基因组挖掘显示存在143个参与渗透适应的基因,解释了其对24%(w/v)NaCl的抗性表型。其中,37个基因组成了完整的异位代谢途径,证实了其在12.5%NaCl胁迫下产生4.38±0.29wt%的异黄酮的能力。一个重要的发现是鉴定了39个负责毒性生物胺组胺整个降解途径的基因,这与在37°C下10天内含有5mM组胺的HA培养基中42.7±2.1%的组胺降解率一致。此外,检测到114个蛋白水解基因和19个脂解基因,这可能有助于其存活以及虾酱的营养品质和风味。值得注意的是,由于其独特的甘氨酸-天冬氨酸-丝氨酸-亮氨酸(GDSL)序列基序,推测的基因vdo2592被发现为可能的新型脂肪酶/酯酶。这是第一份揭示与女性食品相关的Virgibacillus的适应性策略和相关生物技术潜力的报告。我们的发现表明V.dokdonensisT4.6是生产发酵虾酱产品的有前途的发酵剂。
    Virgibacillus spp. stand out as a potent starter culture for accelerating the fermention of fish sauces and shrimp pastes. However, the underlying molecular mechanisms responsible for their adaptation and biotechnological potential remain elusive. Therefore, the present study focuses on phenotypic and genomic analyses of a halophilic bacterium Virgibacillus dokdonensis T4.6, derived from Vietnamese high-salt fermented shrimp paste. The draft genome contained 4,096,868 bp with 3780 predicted coding sequences. Genome mining revealed the presence of 143 genes involved in osmotic adaptation explaining its resistant phenotype to 24% (w/v) NaCl. Among them, 37 genes making up the complete ectoine metabolism pathway, confirmed its ability to produce 4.38 ± 0.29 wt% ectoine under 12.5% NaCl stress. A significant finding was the identification of 39 genes responsible for an entire degradation pathway of the toxic biogenic amine histamine, which was in agreement with its histamine degradation rate of 42.7 ± 2.1% in the HA medium containing 5 mM histamine within 10 days at 37 °C. Furthermore, 114 proteolytic and 19 lipolytic genes were detected which might contribute to its survival as well as the nutrient quality and flavor of shrimp paste. Of note, a putative gene vdo2592 was found as a possible novel lipase/esterase due to its unique Glycine-Aspartate-Serine-Leucine (GDSL) sequence motif. This is the first report to reveal the adaptative strategies and related biotechnological potential of Virgibacillus associated with femented foods. Our findings indicated that V. dokdonensis T4.6 is a promising starter culture for the production of fermented shrimp paste products.
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  • 文章类型: Journal Article
    目标。通过定量脂肪生成和脂肪分解基因的表达,评估富含多酚的橄榄提取物对medaka鱼脂质代谢的可能影响。材料和方法。将成虫medaka鱼在水箱中保持五天,其中五种提取物在水中含量为0.01%,通过富含碳水化合物的饮食导致肥胖,对照组保持在正常饮食的水中。提取物含有7至116毫克/克之间的多酚(橄榄苦苷,羟基酪醇)的抗氧化能力为2-13mmol的2,4,6-三(2-吡啶基)-1,3,5-三嗪/100g。五天后,处死鱼,并通过逆转录提取肝脏mRNA及其互补DNA。通过实时PCR对三个脂肪分解基因和三个脂肪生成基因的互补DNA进行定量。从参照对照组的扩增曲线计算相对基因表达。结果。参与脂解的基因的表达,包括过氧化物酶体增殖物激活受体-±,酰基辅酶A氧化酶1和肉碱棕榈酰转移酶1在进行致胖饮食的鱼类中明显减少,这种情况在富含多酚提取物的鱼类中无法逆转。相比之下,脂肪生成脂肪酸合成酶,乙酰辅酶A羧化酶1和固醇调节元件结合蛋白1基因随着肥胖饮食的增加而大大增加,而橄榄提取物则恢复到正常状态。效果不依赖于总多酚含量,特定的橄榄苦苷或羟基酪醇浓度,或者抗氧化能力,表明有协同效应。结论。橄榄多酚,作为抗生脂剂,对脂质代谢有积极的影响,但是根据提取物,它们在每个基因中的机制是不同的,它支持每种提取物中不同比例的多酚和伴随的植物化学物质的协同机制。
    Aims. To assess the possible effect of polyphenol-rich olive extracts on lipid metabolism in medaka fish by quantifying the expression of lipogenic and lipolytic genes. Materials and methods. Adult medaka fish were maintained in tanks for five days with five extracts at 0.01% in water, causing obesity through a diet rich in carbohydrates, with a control group maintained in water with a normal diet. The extracts contained polyphenols ranging between 7 and 116 mg/g (oleuropein, hydroxytyrosol) with an antioxidant power of 2-13 mmol of 2,4,6-tri(2-pyridyl)-1,3,5-triazine/100 g. After five days, the fish were sacrificed and the hepatic mRNA and its complementary DNA were extracted by reverse transcription. Complementary DNAs were quantified for three lipolytic and three lipogenic genes by real-time PCR. The relative gene expression was calculated from the amplification curves in reference to the control group. Results. The expression of genes involved in lipolysis, including peroxisome proliferator-activated receptor-±, acyl-CoA oxidase 1, and carnitine palmitoyltransferase 1, were clearly decreased in fish subjected to an obesogenic diet, and this situation could not be reversed in fish maintained with polyphenol-rich extracts. In contrast, lipogenic fatty acid synthase, acetyl-CoA carboxylase 1, and sterol regulatory element-binding protein 1 genes increased considerably with the obesogenic diet and reverted to the normal state with the olive extracts. The effect was not dependent on the total polyphenol content, the specific oleuropein or hydroxytyrosol concentration, or the antioxidant power, suggesting a synergistic effect. Conclusion. Olive polyphenols, acting as anti-lipogenic agents, have a positive effect on lipid metabolism, but their mechanism in each gene is different according to the extract, which supports synergistic mechanisms with the different proportions of polyphenols and accompanying phytochemicals in each extract.
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  • 文章类型: Journal Article
    OBJECTIVE: Investigate the effects of moderate continuous aerobic exercise (MCAE) on the inflammatory cytokine profile and expression of lipolytic and thermogenic genes in β1-AR-/- mice adipose tissue.
    METHODS: Four- to five-month-old male wild type (WT) and β1-AR-/- mice were divided into groups: WT control (WTc) and trained (WTt); and β1-AR-/- control (β1-AR-/-c) and trained (β1-AR-/-t). Animals from trained groups were submitted to a MCAE regimen (60 min/day; 60% of maximal speed, 5 days/week) on a treadmill, for 8 weeks. After euthanasia, white epididymal (eWAT) and inguinal (iWAT) and brown (BAT) adipose tissues were dissected and used to determine: adiposity index; adipocyte histomorphometry; cytokine concentration; and gene expression. The content of fat, protein and water of the empty carcass was determined.
    RESULTS: MCAE reduced body weight, fat mass as well as iWAT and BAT adipocyte area in β1-AR-/- animals. Aerobic exercise also diminished the concentrations of pro-inflammatory (IL-12p70, TNF-α, IL-6) and anti-inflammatory (IL-10) cytokines in adipose tissue (iWAT, eWAT or BAT) of β1-AR-/- mice. However, MCAE had no effect on the expression lipolytic and thermogenic genes in β1-AR-/- mice adipose tissue.
    CONCLUSIONS: Alongside reductions in body weight, fat mass and adipocyte area eight weeks of MCAE improves the profile of inflammatory cytokines in β1-AR-/- mice adipose tissue, despite no change in Lipolytic and thermogenic gene expression.
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  • 文章类型: Journal Article
    进行了为期10周的饲喂试验,以评估不同饲粮大豆油(SO)水平对生长性能的影响。内脏中的脂肪酸组成和脂质沉积,肝脏的组织学和组织化学,泥浆鱼幼鱼肠道和肝脂代谢相关基因的表达。泥电动机(平均值±s.d.质量0·40±0·01g)饲喂了五种含有不同浓度SO的实验饮食:0、20、32、56和100%SO和含有100%鱼油的饮食(100%FO)。饲喂20%SO和100%FO日粮的根茎的质量增加和比生长速率显着高于其他组(P<0·05)。随着饲料SO水平的增加,内脏的脂质含量和肝脏中细胞质空泡的数量增加。同时,增加饮食SO水平上调脂肪生成基因的信使(m)RNA水平(如Δ6fad,scd,pparγ,fas和srebp-1)并下调脂解基因的mRNA水平(例如pparα,肝脏中的cpt1,atgl和hsl)。20:4n-6的百分比随着饲粮SO水平的增加而显著增加(P<0.05),这可能与Δ6fadmRNA水平的上调有关。最高水平的饮食SO,然而,对增长绩效有负面影响,内脏的脂质沉积以及肝脏和肠道的组织学和组织化学。饮食SO水平增加引起的脂质积累增加可能是由于脂解与脂肪生成之间以及该物种脂质的出口与进口之间的竞争而通过不同的脂质代谢策略发生的。
    A 10 week feeding trial was conducted to evaluate the effects of different dietary soybean oil (SO) levels on growth performance, fatty-acid composition and lipid deposition in viscera, histology and histochemistry of liver, intestine and hepatic-lipid metabolism-related gene expressions in pond loach Misgurnus anguillicaudatus juveniles. Misgurnus anguillicaudatus (mean ± s.d. mass 0·40 ± 0·01 g) were fed five experimental diets containing SO at different concentrations: 0, 20, 32, 56 and 100% SO and a diet containing 100% fish oil (100% FO). The mass gains and specific growth rates of M. anguillicaudatus fed 20% SO and 100% FO diets were significantly higher than those of the other groups (P < 0·05). The lipid content of viscera and the amount of cytoplasmic vacuolation in the liver increased with incremental dietary SO level. Meanwhile, increasing dietary SO levels up-regulated the messenger (m)RNA levels of lipogenic genes (such as Δ6fad, scd, pparγ, fas and srebp-1) and down-regulated the mRNA levels of the lipolytic genes (such as pparα, cpt1, atgl and hsl) in the liver. The percentage of 20:4n-6 significantly (P < 0·05) increased with increasing dietary SO level, which might be correlated with the up-regulation of the mRNA level of Δ6fad. The highest levels of dietary SO, however, had a negative effect on growth performance, lipid deposition of viscera and histology and histochemstry of liver and intestine. The increased lipid accumulation induced by incremental dietary SO level probably occurred through different strategies for lipid metabolism as a result of competition between lipolysis and lipogenesis and between export and import of lipids in this species.
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  • 文章类型: Journal Article
    BACKGROUND: Specific bio-active dietary compounds modulate numerous metabolic processes in adipose tissue (AT), including pre-adipocyte proliferation and differentiation. AT dysfunction, rather than an increased fat mass per se, is strongly associated with the development of insulin resistance and is characterized by impaired adipogenesis, hypertrophic adipocytes, inflammation, and impairments in substrate metabolism. A better understanding of mechanisms underlying AT dysfunction may provide new strategies for the treatment of obesity-associated metabolic diseases. Here we evaluated the role of (all-E)-lycopene (Lyc), eicosapentaenoic acid (EPA) or trans-resveratrol (Res) and combinations thereof on human white adipocyte function.
    METHODS: In-vitro differentiating human pre-adipocytes were treated with EPA, Lyc and Res or their combinations for 14 days. The effects on intracellular lipid droplet (LD) accumulation, secreted anti- and pro-inflammatory cyto-/adipokines (e.g. adiponectin, IL-6, IL-8/CXCL-8 and MCP-1/CCL2) and on gene expression of markers of adipocyte differentiation and substrate metabolism (e.g. PPAR-gamma, C/EBP-alpha, GLUT-4, FAS, ATGL, HSL, and PLIN-1) were measured by fluorescent microscopy (Cellomics™), multi-parametric LiquiChip® technology and quantitative RT-PCR, respectively.
    RESULTS: Treatment of differentiating adipocytes for 14 days with the combination of Lyc/Res and EPA/Res resulted in significantly inhibited LD formation (~ -25 and -20%, respectively) compared to the effects of the single compounds. These morphological changes were accompanied by increased mRNA levels of the adipogenic marker PPAR-gamma and the lipase ATGL and by decreased expression levels of lipogenic markers (LPL, FAS, GLUT-4) and the LD-covering protein PLIN-1. In addition, a blunted adipocyte secretion of pro-inflammatory cytokines (IL-6 and MCP-1) and adiponectin was observed following treatment with these compounds.
    CONCLUSIONS: The combination of the dietary bio-actives Lyc and EPA with Res might influence adipocyte function by affecting the balance between adipogenic, lipogenic and lipolytic gene expression, resulting in a reduced LD storage and a less inflammatory secretion profile. Taken together, our results indicate that combinations of dietary compounds may be beneficial for the prevention and treatment of metabolic disorders via effects on human white adipocyte function.
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