Cyclodepsipeptides: Isolation from Endophytic Fungi of Sarcophyton ehrenbergi and Verification of Their Larvicidal Activity via In-Vitro and In-Silico Studies

Culex pipiens mosquitoes are vectors to many viruses and can transmit diseases such as filariasis and avian malaria. The present study evaluated the larvicidal activity of marine-derived endophytic fungi Aspergillus nomius and Aspergillus flavus from the soft coral Sarcophyton ehrenbergi along with two known cyclodepsipeptide compounds, scopularide A (1) and B (2), isolated from A. flavus extract, against third-instar larvae of C. pipiens, using distilled water as a negative control and toosenedanin as a positive control. The structures of the isolated compounds were confirmed by various spectroscopic analyses. The lethal concentrations (LC₅₀ and LC₉₀) were calculated by probit analysis. Scopularide A was the most potent after 96 h treatment, with LC₅₀ and LC₉₀ values of 58.96 and 994.31 ppm, respectively, and with 82.66% mortality at a concentration of 300 ppm. To unravel the biochemical mechanism of the tested extracts and compounds, their effects against protease, chitinase, phenoloxidases and lipase enzymes from the whole-body tissue of C. pipiens were evaluated after 72 h treatment at LC₅₀ dose. Superior activity was observed for A. flavus extract against all tested enzymes. A molecular docking study was conducted for scopularide A and B on the four tested enzymes, to further verify the observed activity. Results revealed good binding affinities for both compounds as compared to the docked ligands, mainly via a number of hydrogen bonds. This was the first study to report the isolation of endophytic fungi A. flavus and A. nomius from the marine soft coral S. ehrenbergi. The endophytic fungal extract of A. flavus was found to be a promising source for a natural larvicidal agent against C. pipiens populations.     

不同淀粉源饲粮对肉仔鸡小肠消化酶活性、盲肠挥发性脂肪酸含量和微生物菌群的影响

本试验旨在研究不同淀粉源饲粮对肉仔鸡小肠消化酶活性、盲肠挥发性脂肪酸含量和微生物菌群的影响.选取1日龄健康爱拔益加(AA)肉仔鸡480只,随机分为5组,每组8个重复,每个重复12只鸡.5组分别饲喂玉米淀粉、豌豆淀粉、木薯淀粉、高粱淀粉和小麦淀粉配制的半纯合等氮等能饲粮,5组饲粮总淀粉水平一致.试验期21 d.结果表明:...     

脂肪酶催化菜籽油制备生物柴油工艺优化

利用脂肪酶LVK在以正己烷为溶剂的体系中催化菜籽油与乙醇酯交换合成生物柴油。为提高酯交换率,采用响应面实验设计和分析方法对菜籽油的酯交换反应条件进行优化,得到最佳工艺条件：醇油摩尔比5．3：1,脂肪酶与油脂的质量比为15％,反应温度40℃,反应时间34．5h,溶剂（正已烷）量18．4％,乙醇一次加入,在此工艺条件下菜籽油的酯交换率达到93．48％。结果表明正己烷体系能很好解决乙醇与菜籽油的互溶性,消除乙醇对脂肪酶的毒害作用。     

微乳液凝胶固定化脂肪酶催化消旋布洛芬辛酯水解反应的研究

初步研究了AOT-正庚烷-明胶微乳液凝胶固定化脂肪酶催化消旋布洛芬辛酯水解反应的影响因素。结果表明:在正庚烷中,布洛芬辛酯水解反应能够顺利进行,产物为光学纯度很高的S-构型布洛芬,转化率为0.3217时,其对映体过量值为0.9826;凝胶R值(水与AOT的摩尔数之比)对酶催化反应有较大影响,最佳R值为73;当酶含量固定,底物浓度为0.10 mol/L时,转化率最大并能得到较高对映体过量值的产物;当反应时间超过16 h时,水解反应达到平衡;固定化酶的重复使用次数与溶剂中的含水量有关,正庚烷中少量的水有助于增加酶的重复使用次数。     

Genome-wide identification and expression analysis of GDSL esterase/lipase genes in tomato

The GDSL esterase/lipase family contains many functional genes that perform important biological functions in growth and development, morphogenesis, seed oil synthesis, and defense responses in plants. The expression of GDSL esterase/lipase genes can respond to biotic and abiotic stresses. Although GDSL esterase/lipase family genes have been identified and studied in other plants, they have not been identified and their functions remain unclear in tomato. This study is the first to identify 80 G...     

儿茶素单体EGC对胰脂肪酶的抑制作用及其机理研究

为研究儿茶素单体表没食子儿茶素（Epigallocatechin,EGC）对胰脂肪酶的抑制作用及机制,以干燥绿茶为原料,采用热水提取法、三氯甲烷脱色素和柱层析分离,得到EGC单体。采用扫描电子显微镜、傅里叶红外光谱、核磁共振氢谱对纯化的EGC单体进行结构表征,再通过滴定法研究EGC对胰脂肪酶的抑制作用及类型,然后通过荧光光谱法以及分子对接技术表征了EGC对胰脂肪酶结构的影响。结果表明,EGC以非竞争性方式对胰脂肪酶表现出抑制作用,并且抑制效果随EGC浓度的增加呈持续上升趋势,半抑制浓度为(1.62±0.085) mg·mL^-1。EGC对胰脂肪酶有荧光猝灭作用,可通过分子间氢键和疏水相互作用与酶中的氨基酸残基结合,导致酶的化学结构和空间构象发生变化,从而降低酶活性。EGC主要通过改变胰脂肪酶的化学结构和空间构象来抑制该酶活性,从而达到降血脂功效。     

谷关假单胞菌脂肪酶基因PgLip1的克隆和表达

低温脂肪酶在饲料、洗涤、有机合成等行业中具有重要的应用价值，然而目前已发现的低温脂肪酶数量不多，对其基因多样性了解不够深入，因此有必要从自然环境中挖掘新的低温脂肪酶基因。为了获得低温脂肪酶基因资源，通过基因组序列比对分析，从谷关假单胞菌（Pseudomonas guguanensis）中发现并克隆获得一个脂肪酶基因P. guguanensis lipase 1 (PgLip1),并对该基因进行序列分析和蛋白质预测，表达后检测PgLip1的最适底物、最适pH、最适温度，同时研究了表面活性剂、变性剂、EDTA、金属离子及有机溶剂对该酶活性的影响。结果显示：该基因片段大小为840 bp，预测编码含279个氨基酸残基的蛋白质，氨基酸序列与来源于门多萨假单胞菌（Pseudomonas mendocina）的脂肪酶相似性92.6%。该酶的最适温度为10 ℃，在0 ℃相对酶活为60.2%，是一个低温脂肪酶；其最适pH为8.5，在pH 3.0~12.0时，仍然具有62%以上的相对酶活。40 ℃处理45 min后，PgLip1还能保持72.6%的残余酶活。该酶的最适反应底物为对硝基苯基丁酸酯(pNPB)；在高浓度异丙醇、异戊醇和乙酸乙酯中的酶活反而高于相应较低浓度中，吐温-20、吐温-80和Triton X-100表面活性剂能较大程度激活PgLip1（3.2~5.9倍）。研究丰富了对低温脂肪酶生物多样性的科学认识，所获得的PgLip1是一个受表面活性剂特异激活且对部分高浓度有机溶剂耐受的低温脂肪酶，具有较好的应用前景。     

Effects of dietary sodium butyrate on growth,digestive enzymes,body composition and nutrient retention-related gene expression of juvenile yellow catfish (Pelteobagrus fulvidraco)

An 8-week feeding trial was conducted to evaluate the effects of sodium butyrate (SB) on growth, digestive enzymes, body composition and nutrient retention-related gene expression of juvenile yellow catfish (Pelteobagrus fulvidraco). Five isonitrogenous and isolipidic diets (420 g/kg protein and 90 g/kg lipid) were formulated to contain 0 (control), 250, 500, 1,000 or 2,000 mg/kg SB. Triplicate groups of 40 fish (BW = 1.26 ± 0.01 g) per tank (300-L cylindrical fiberglass tanks) for each diet were fed to apparent satiation twice daily. Stomach, hepatopancreas and intestine samples were obtained for digestive enzymes activities analyses. A real-time quantitative PCR analysis was performed to determine the relative expression of target of rapamycin (TOR) and lipoprotein lipase (LPL) in the hepatopancreas and intestine. Fish fed the diets supplemented with SB at 500 and 1,000 mg/kg showed significantly higher specific growth rate and significantly lower feed conversion ratio compared to the control (P < 0.05). Dietary SB inclusion did not alter activities of intestinal amylase, creatine kinase and sodium–potassium adenosine triphosphatase (Na⁺/K⁺-ATPase), but increased activities of hepatic trypsin, stomachic lipase, intestinal lipase, alkaline phosphatase and γ-glutamyl transpeptidase for fish fed 1,000 mg/kg SB compared to the control (P < 0.05). Intestine length index, intestine somatic index, fold height and muscular thickness of distal intestine were significantly higher in 1,000 mg/kg SB groups compared to the control (P < 0.05). Significantly higher levels of whole-body crude protein, ash, calcium, phosphorus, nutrition retention and relative mRNA of intestinal TOR were observed in 1,000 mg/kg SB group (P < 0.05). Whole-body lipid content and hepatopancreas LPL mRNA expression in 2,000 mg/kg SB group were significantly higher than the control (P < 0.05). Relative mRNA levels of intestinal LPL and hepatopancreas TOR were significantly higher in the 500 mg/kg SB group compared to those in other groups (P < 0.05). The increased growth performance, digestive enzymes and nutrient retention in fish fed the diets supplemented with SB at 500 and 1,000 mg/kg suggests that SB can be a desirable growth promoter as an antibiotic alternative in diets.     

蒙古冰草GDSL酯酶/脂肪酶基因的序列及表达分析

为挖掘蒙古冰草(Agropyron mongolicum)中的GDSL酯酶/脂肪酶基因，以干旱胁迫下蒙古冰草转录组测序得到的Unigenes为对象，利用生物信息学方法对蒙古冰草GDSL脂肪酶基因进行理化性质、亚细胞定位、同源性以及系统发育进化分析，同时对不同干旱处理条件下的GDSL脂肪酶基因进行表达分析。结果表明：蒙古冰草中52个GDSL脂肪酶基因，蛋白质分子量在6.80～44.77 kU,66～420个氨基酸，pI 4.27～9.42;52个蒙古冰草GDSL脂肪酶基因亚细胞定位预测显示细胞外基质、叶绿体、细胞核、细胞质、细胞膜、细胞壁均有分布；将其与13个已知具有抗旱功能的GDSL脂肪酶基因进行同源性对比及进化树构建，蒙古冰草GDSL脂肪酶基因与水稻、大麦、拟南芥、木豆、大豆、蜡梅、辣椒的GDSL脂肪酶基因均有较高相似性。在干旱处理0.5、1.0、2.0、3.0、5.0、7.0 d及复水1.0 d,与对照(CK)相比，14个基因呈显著下调表达，38个基因呈现为上调表达。对6个蒙古冰草GDSL脂肪酶蛋白跨膜及信号肽和磷酸化预测结果显示，DN26944＿c0＿g1属于跨膜蛋白；DN269...     

产脂肪酶菌株的筛选、鉴定及响应面法优化发酵条件

从土壤中分离纯化出高产脂肪酶的菌株,通过在360nm紫外光下比较罗丹明B(Rhodamine B)筛选培养基中荧光圈的大小后得到7株菌株。利用对硝基苯酚-分光光度法测定目的菌株的酶活大小对菌株进行复筛,最终得到一株酶活较高的菌株Youzh-1,经细菌形态观察,16S rRNA序列分析,确定其为荧光假单胞菌(Pseudomonas fluorescens strain ORTB3)。对此菌株进行发酵条件的优化,首先对发酵条件进行单因素的优化,在单因素实验的基础上对Youzh-1设计Box-Behnken实验进一步优化培养基最终得到最优发酵条件为葡萄糖18.825g/L、牛肉膏25.98g/L、,氯化钠10g/L、硫酸镁0.1g/L、磷酸二氢钾0.5g/L、橄榄油乳化液12ml/ml,在温度41℃、pH6的条件下进行发酵实验验证,经过测定发现其脂肪酶粗酶酶活为79.87U/ml,相比初始酶活37.67U/ml提高了112%。这为脂肪酶的工业化生产提供了理论依据。