脂肪酶修饰丝网印刷电极测定三油酸甘油酯

制备了用于快速检测三油酸甘油酯的脂肪酶修饰丝网印刷电极（SPE）电化学传感器单元。用循环伏安法（CV）和计时电流法（CC）研究了三油酸甘油酯在脂肪酶修饰SPE上的电化学行为,确定了测定甘油酯的最优条件,建立了测定三油酸甘油酯浓度的电化学方法。其中CV法的线性范围为0.01~5g?L-1,检出限为2.5mg?L-1;CC法的线性范围为0.001~550g?L-1,检出限为0.2mg?L-1。以CV法考察样品的加标回收率,6个葵花籽油样品的加标回收率为94%~105%。     

乙烯对脂肪酶活力的直接作用及机理初探

研究了乙烯对脂肪酶活力的直接作用及其机理. 结果表明: 低浓度乙烯能使脂肪酶催化三油酸甘油酯的水解活力提高; 当乙烯浓度为0.9834 mmol•L^－1时, 酶活力提高13.0%. 高浓度乙烯降低脂肪酶活力; 当乙烯浓度为7.9669 mmol•L^－1时, 酶活力下降24.5%. 加入乙烯的酶最适温度向高温偏移10～15 ℃, 而酶的最适pH值不变. 在pH＝7.9时, 乙烯使酶活力升高较大, pH为4.5～7.5, 8.5, 9.5～11时酶的活力降低. 加入乙烯的酶与对照相比, 其紫外吸收和荧光发射强度均有较大幅度增加, 荧光偏振度、比旋光度和粘度显著下降. DSC分析表明: 在低温范围内酶的可逆吸热峰值温度明显高于对照, 而热焓变低于对照; 在高温范围内酶的不可逆吸热峰值温度和热焓变都低于对照. 这些结果证实了乙烯可以直接影响酶的微环境和构象. 乙烯对脂肪酶的直接作用机制可能是通过改变酶的微环境以及渗入到酶分子内部改变酶构象而引起酶活力的改变.     

乙烯对脂肪酶活力的直接作用及机理初探

研究了乙烯对脂肪酶活力的直接作用及其机理. 结果表明: 低浓度乙烯能使脂肪酶催化三油酸甘油酯的水解活力提高; 当乙烯浓度为0.9834 mmol•L^－1时, 酶活力提高13.0%. 高浓度乙烯降低脂肪酶活力; 当乙烯浓度为7.9669 mmol•L^－1时, 酶活力下降24.5%. 加入乙烯的酶最适温度向高温偏移10～15 ℃, 而酶的最适pH值不变. 在pH＝7.9时, 乙烯使酶活力升高较大, pH为4.5～7.5, 8.5, 9.5～11时酶的活力降低. 加入乙烯的酶与对照相比, 其紫外吸收和荧光发射强度均有较大幅度增加, 荧光偏振度、比旋光度和粘度显著下降. DSC分析表明: 在低温范围内酶的可逆吸热峰值温度明显高于对照, 而热焓变低于对照; 在高温范围内酶的不可逆吸热峰值温度和热焓变都低于对照. 这些结果证实了乙烯可以直接影响酶的微环境和构象. 乙烯对脂肪酶的直接作用机制可能是通过改变酶的微环境以及渗入到酶分子内部改变酶构象而引起酶活力的改变.     

Using Lipase Modified Screen-printed Electrode as Glyceryltrioleate Biosensor

制备了用于快速检测三油酸甘油酯的脂肪酶修饰丝网印刷电极(SPE)电化学传感器单元.用循环伏安法(CV)和计时电流法(CC)研究了三油酸甘油酯在脂肪酶修饰SPE上的电化学行为,确定了测定甘油酯的最优条件,建立了测定三油酸甘油酯浓度的电化学方法.其中CV法的线性范围为0.01～5 g,L-1,检出限为2.5 mg· L-1; CC法的线性范围为0.001～5.50 g·L-1,检出限为0.2 mg· L-1.以CV法考察样品的加标回收率,6个葵花籽油样品的加标回收率为94％ ～ 105％.     

AEOT反胶束中脂肪酶的催化活性

反胶束已广泛应用于膜模拟化学和蛋白质的液 液萃取中[1～ 3] ,反胶束酶反应作为实现有机相酶催化的方法之一 ,具有许多独特的优点 ,反胶束独特的结构特征使表面活性剂分子组成的膜将油水相隔开 ,从而有利于保持酶的活性和稳定性。酶在反胶束的微水环境中比在水溶液中更接近天然的细胞内环境 ,在这里酶和底物分子均可得到有效的分散 ,接触几率大大提高 ,因而催化效率也得到很大提高。反胶束可以适用于各种类型的 (亲水的、疏水的和双亲的 )底物[4] ,已逐步形成“胶束酶学”的研究分支 ,研究胶束酶学的Martinek等[3] 曾预言 :反胶束体系有可…     

基于尼古丁对胆碱氧化酶的抑制作用测定尼古丁的研究

基于尼古丁对胆碱氧化酶(CHOD)的抑制作用,将胆碱氧化酶电极用于微量尼古丁的测定。酶电极制作中,先在铂电极表面修饰一层壳聚糖膜,再用戊二醛交联CHOD。采用这种酶固定化方法,电极在4. 0×10-6 ~ 3×10-3 mol/L胆碱的浓度范围呈线性关系。探讨了工作电位、pH、底物浓度等实验条件对酶电极性能及抑制过程中响应电流的影响,测定了电极的重现性、干扰及使用寿命。电极检测尼古丁的线性范围为1. 5×10-5 ~3×10-3 mol/L;检出限为1. 25×10-5 mol/L。将电极用于实际样品烟草中尼古丁回收率的测定,结果良好。     

尼古丁与DNA相互作用的电化学研究

用线性扫描伏安法、循环伏安法及光谱法研究了尼古丁与DNA在0.2 mol/LpH 6.0的H2C2O4缓冲溶液中的相互作用。研究结果显示,随着DNA的加入,尼古丁峰电流降低,峰电位正移,说明尼古丁是以嵌入形式与DNA结合,生成了一种结合比为2∶1的非电活性化合物,结合常数β为1.56×109。并用多种电化学方法求得了电极过程的动力学参数。     

华根霉全细胞脂肪酶催化合成油酸油醇酯

 比较了10种不同来源的脂肪酶催化油酸与油醇酯化合成油酸油醇酯的能力,其中华根霉Rhizopus chinensis CCTCC M201021全细胞脂肪酶的催化能力最强,其反应转化率可达到90%以上. 酯化反应的最佳油酸/油醇底物摩尔比为1.5, 最佳油酸浓度为0.3 mol/L, 细胞干粉含水量为3.0%～7.5%时对酯化反应最为有利. 以生物相容性指数logP值为指标选择不同的有机溶剂作为有机相进行酯化反应,发现logP值为3.5～4.5的有机溶剂促进酯化效果较好. 全细胞脂肪酶的pH适应范围很广,最佳pH在9.0左右; 最佳反应温度为30 ℃.     

尼古丁的“口供”

以口供的形式介绍了尼古丁的分子结构和部分生理功能,通过真实案件揭示尼古丁对人体的毒性。结合医学知识,阐述了尼古丁中毒的过程、吸烟上瘾的机理和尼古丁对人体的损害,并对人类健康生活提出禁烟建议。     

反相胶束中脂肪酶催化猪油的甘油解反应

单脂肪酸甘油酯;反相胶束中脂肪酶催化猪油的甘油解反应     

Lipase immobilized on hydrophobic microporous polypropylene for the hydrolysis of palm kernel olein

Lipase (triacylglycerol ester hydrolase, EC 3.1.1.3) fromRhizopus arrhizus was immobilized in this work by adsorption on microporous polypropylene and employed for the lipolysis of palm kernel olein. The optimum operating temperature for the lipolysis reaction was determined. The reaction follows Michaelis-Menten kinetics with product competitive inhibition for substrate concentrations in the range of 0.175–0.877M. The apparentK _m and V_max were 0.42M and 691 U/mg protein, respectively. A dissociation constant of the enzymeproduct complex,K _I = 29.73 mM, for the product inhibition was also determined. Additionally, the time-courses of the reaction for various substrate concentrations were obtained and correlated sufficiently with those predicted from the theoretical rate equation for a period of up to 2 h. Experimental results indicated that discrepancies between the observed results and the predicted ones increase with reaction time.     

15-羟基十五烷酸的脂肪酶催化合成环十五内酯

15-羟基十五烷酸的脂肪酶催化合成环十五内酯     

Free Linoleic Acid and Oleic Acid Reduce Fat Digestion and Absorption In Vivo as Potent Pancreatic Lipase Inhibitors Derived from Sesame Meal

Pancreatic lipase catalyzes the cleavage of triacylglycerols at the oil–water interface, and is known as the dominant determiner of dietary fat digestion. Reducing dietary fat digestion and absorption by modulating the activity of pancreatic lipase has become a favorable strategy to tackle obesity. Orlistat is, at present, the only pancreatic lipase inhibitor approved for the treatment of obesity; however, an array of gastrointestinal adverse effects associated with orlistat limits its tolerability. As a safe alternative to orlistat, a number of natural product-derived compounds with varying degrees of pancreatic lipase inhibitory activity have been reported. We herein reported that bioactivity-guided fractionation of sesame meal led to the identification of free linoleic acid and oleic acid as potent inhibitors of porcine pancreatic lipase in vitro with an IC50 of 23.1 µg/mL (82.4 µM) and 11.7 µg/mL (41.4 µM), respectively. In rats, a single oral dose of the mixture of these fatty acids significantly suppressed the elevation of blood triacylglycerol level following fat intake. These results substantiate the role of free linoleic acid and oleic acid as a novel class of natural product-derived functional molecules that act as pancreatic lipase inhibitors, and their potential for healthy, routine-based weight management.     

A Reversible and Selective Inhibitor of Monoacylglycerol Lipase Ameliorates Multiple Sclerosis

Monoacylglycerol lipase (MAGL) is the enzyme responsible for the inactivation of the endocannabinoid 2‐arachidonoylglycerol (2‐AG). MAGL inhibitors show analgesic and tissue‐protecting effects in several disease models. However, the few efficient and selective MAGL inhibitors described to date block the enzyme irreversibly, and this can lead to pharmacological tolerance. Hence, additional classes of MAGL inhibitors are needed to validate this enzyme as a therapeutic target. Here we report a potent, selective, and reversible MAGL inhibitor (IC₅₀=0.18 μM ) which is active in vivo and ameliorates the clinical progression of a multiple sclerosis (MS) mouse model without inducing undesirable CB₁‐mediated side effects. These results support the interest in MAGL as a target for the treatment of MS.     

Enhancing the Photo and Thermal Stability of Nicotine through Crystal Engineering with Gentisic Acid

The use of crystal engineering to convert liquids into crystalline solids remains a powerful method for inhibiting undesired degradation pathways. When nicotine, a liquid sensitive to both light and air, is combined with the GRAS-listed compound, gentisic acid, the resulting crystalline solid, exhibits enhanced photo and thermal stability. Despite a modest ΔT_m of 42.7 °C, the melting point of 155.9 °C for the nicotinium gentisate salt is the highest reported for nicotine-containing crystalline solids. An analysis of the crystal packing and thermodynamic properties provides context for the observed properties.     

壳聚糖固定化碱性脂肪酶的研究

以蟹壳为原料提取壳聚糖,用戊二醛作交联剂,将碱性脂肪酶固定于壳聚糖上。同时探讨了一定量干壳聚糖载体与交联剂浓度、给酶量等关系的最适固定化酶条件,并对固定化酶的热稳定性、操作稳定性、米氏常数、最适温度、离子强度的影响以及使用半衰期等理化性质进行探讨。     

High-Resolution Bioassay Profiling with Complemented Sensitivity and Resolution for Pancreatic Lipase Inhibitor Screening

How to rapidly and accurately screen bioactive components from complex natural products remains a major challenge. In this study, a screening platform for pancreatic lipase (PL) inhibitors was established by combining magnetic beads-based ligand fishing and high-resolution bioassay profiling. This platform was well validated using a mixture of standard compounds, i.e., (-)- epigallocatechin gallate (EGCG), luteolin and schisandrin. The dose–effect relationship of high-resolution bioassay profiling was demonstrated by the standard mixture with different concentrations for each compound. The screening of PL inhibitors from green tea extract at the concentrations of 0.2, 0.5 and 1.0 mg/mL by independent high-resolution bioassay profiling was performed. After sample pre-treatment by ligand fishing, green tea extract at the concentration of 0.2 mg/mL was specifically enriched and simplified, and consequently screened through the high-resolution bioassay profiling. As a result, three PL inhibitors, i.e., EGCG, (-)-Gallocatechin gallate (GCG) and (-)-Epicatechin gallate (ECG), were rapidly identified from the complex matrix. The established platform proved to be capable of enriching affinity binders and eliminating nonbinders in sample pre-treatment by ligand fishing, which overcame the technical challenges of high-resolution bioassay profiling in the aspects of sensitivity and resolution. Meanwhile, the high-resolution bioassay profiling possesses the ability of direct bioactive assessment, parallel structural analysis and identification after separation. The established platform allowed more accurate and rapid screening of PL inhibitors, which greatly facilitated natural product-based drug screening.     

Discovery of Novel Epoxyketone Peptides as Lipase Inhibitors

Obesity is the most common nutritional disorder in the developed world and is associated with important comorbidities. Pancreatic lipase (PL) inhibitors play a key role in the metabolism of human fat. A series of novel epoxyketones peptide derivatives were investigated for their pancreatic lipase inhibitory activity. The epoxyketone moiety is a well-known reactive electrophile group that has been used as part of proteasome inhibitors in cancer therapy, and it is widely believed that these are very selective for targeting the proteasome active site. Here we investigated various peptide derivatives with an epoxide warhead for their anti-lipase activity. The assessment of these novel epoxyketones was performed by an in-house method that we developed for rapid screening and identification of lipase inhibitors using GC-FID. Herein, we present a novel anti-lipase pharmacophore based on epoxyketone peptide derivatives that showed potent anti-lipase activity. Many of these derivatives had comparable or more potent activity than the clinically used lipase inhibitors such as orlistat. In addition, the lipase appears to be inhibited by a wide range of epoxyketone analogues regardless of the configuration of the epoxide in the epoxyketone moiety. The presented data in this study shows the first example of the use of epoxyketone peptides as novel lipase inhibitors.