Inhibition of lipase from rice (Oryza sativa) by diethyl-p-nitrophenyl phosphate

The inhibition of rice bran lipase (RBL) by diethyl-p-nitrophenyl phosphate was studied with reference to kinetics, nature of inhibition and also elucidate the effect of the inhibitor on the structure—function of the enzyme. Enzyme activity measurements shows that the inhibitor is more effective at 0.050 mM concentration of diethyl-p-nitrophenyl phosphate and the activity is 50% at this level of inhibitor concentration. The affinity of substrate for the enzyme was observed by the increase in the velocity of the reaction with increase in the substrate concentrations and double reciprocal plot indicates that the inhibition followed a competitive in nature and inhibition constant K _i is found to be 0.016 mM at pH 7.0. The decrease in apparent thermal denaturation temperature to 4 °C compared to control indicates the destabilization of enzyme in the presence of inhibitor. Fluorescence spectral measurements suggests that pronounced quenching of fluorescence intensity of RBL occurs at higher concentrations of diethyl-p-nitrophenyl phosphate and ‘K _a’ value was found to be 2.4 × 10⁴ M⁻¹ with free energy change ΔG^o—26 kJ/mol at 30 °C suggesting strong binding between the enzyme and the inhibitor with microenvironmental changes occur at the active site or in the neighbourhood of active site. The far UV-CD data suggest that there is no significant changes in the conformation of the enzyme as a result of binding of diethyl-p-nitrophenyl phosphate. These results indicate that diethyl-p-nitrophenyl phosphate is a inhibitor of RBL and binds to the enzyme in brining about inhibition without any structural alterations.     

矢车菊素-3-葡萄糖苷对骨骼肌细胞脂蛋白脂肪酶活性的影响

富含矢车菊素-3-葡萄糖苷(Cy-3-g)的花色苷提取物可抑制肥胖,但其作用机制不明。骨骼肌细胞脂蛋白脂肪酶(LPL)活性与肥胖相关,LPL使骨骼肌中的β-脂肪酸氧化增强,从而加速清除体内循环的甘油三酯(TG),抑制TG流向脂肪组织积聚而导致肥胖。本实验建立稳定的骨骼肌细胞分离培养方法,通过花色苷Cy-3-g干预骨骼肌细胞,研究Cy-3-g对细胞LPL活性的影响及其作用机制。结果表明：Cy-3-g通过激活一磷酸腺苷激活的蛋白激酶(AMPK)上调骨骼肌细胞LPL活性,提示Cy-3-g具有潜在的调节机体脂代谢作用,与含Cy-3-g的花色苷提取物的肥胖抑制作用密切相关。     

酶法动态动力学拆分制备R-(-)-乙酰基邻氯扁桃酸

采用假单胞菌脂肪酶Pseudomonas sp. ECU1011催化乙酰基邻氯扁桃酸进行不对称水解,利用突变后的扁桃酸消旋酶(V29I)对拆分后的产物S-(-)-邻氯扁桃酸进行消旋,消旋后的邻氯扁桃酸经过酰化重新被利用到水解反应中,实现了酶法动态动力学拆分制备R-(-)-乙酰基邻氯扁桃酸。通过对拆分反应、拆分混合物的分离回收以及消旋反应的工艺优化,最终获得光学纯度ee>99.9%的R-(-)-乙酰基邻氯扁桃酸,其收率达80%。本研究建立的R-(-)-乙酰基邻氯扁桃酸的动态动力学拆分工艺,对其工业化应用具有重要的指导意义。     

酶法富集小黄鱼内脏鱼油中EPA和DHA甘油酯

目的：以小黄鱼内脏精炼鱼油为原料,通过脂肪酶选择性水解法富集二十碳五烯酸（eicosapentaenoic acid,EPA）甘油酯和二十二碳六烯酸（docosahexaenoic acid,DHA）甘油酯。方法：采用气相色谱峰面积外标法定量测定EPA和DHA甘油酯总含量。通过单因素和响应面试验,考察反应时间、pH值、反应温度、加酶量等因素对富集效果的影响。结果：确定最佳工艺条件为反应时间4 h、pH 8.0、反应温度30 ℃、加酶量1.0%,在此条件下,EPA和DHA甘油酯总含量为21.65%,且4 个因素对EPA和DHA甘油酯富集的影响依次增强。结论：富集后鱼油理化指标和感官评价均优于精炼鱼油,EPA和DHA甘油酯总含量是富集前的1.74 倍,获得了天然的EPA和DHA甘油酯。     

介孔材料SBA-15固定化脂肪酶的研究进展

介孔材料由于其孔道分布有序且孔径均匀等优点而在固定化酶催化领域引起人们的广泛关注.本文综述了新型介孔材料SBA-15对脂肪酶固定化的研究进展.总结了SBA-15的孔径大小、形貌及等电点等因素对脂肪酶固定化的影响.归纳了SBA-15上三种不同固定化方法的优缺点,并介绍了SBA-15固定化脂肪酶在手性拆分、酯水解、酯合成及转醇化反应等领域的应用.最后提出SBA-15固定化脂肪酶在发展过程中存在的问题以及今后的发展趋势.     

脂肪酶催化动力学拆分大分子手性化合物的研究进展

脂肪酶已广泛用于制备手性化合物的动力学拆分中,但大多数酶催化拆分外消旋化合物,特别是大分子手性化合物的对映选择性不很理想。目前,提高脂肪酶对映选择性的研究主要在改造脂肪酶蛋白结构、优化体系的反应条件、改善反应过程以及对映选择性抑制等方面进行。本文主要介绍了几种改善脂肪酶催化动力学拆分大分子手性化合物对映选择性的合理方法。     

Lipase-catalyzed Synthesis of Caffeic Acid Phenethyl Ester in Ionic Liquids： Effect of Specific Ions and Reaction Parameters

Caffeic acid phenethyl ester（CAPE） is a rare, naturally occurring phenolic food additive. This work systematically reported fundamental data on conversion of caffeic acid（CA）, yield of CAPE, and reactive selectivity during the lipase-catalyzed esterification process of CA and phenylethanol（PE） in ionic liquids（ILs）. Sixteen ILs were selected as the reaction media, and the relative lipase-catalyzed synthesis properties of CAPE were measured in an effort to enhance the yield of CAPE with high selectivity. The results indicated that ILs containing weakly coordinating anions and cations with adequate alkyl chain length improved the synthesis of CAPE. [Emim][Tf2 N] was selected as the optimal reaction media. The optimal parameters were as follows by response surface methodology（RSM）： reaction temperature, 84.0 °C; mass ratio of Novozym 435 to CA, 14︰1; and molar ratio of PE to CA, 16 ︰ 1. The highest reactive selectivity of CAPE catalyzed by Novozym 435 in [Emim][Tf2 N] reached 64.55%（CA conversion 98.76% and CAPE yield 63.75%, respectively）. Thus, lipase-catalyzed esterification in ILs is a promising method suitable for CAPE production.     

基于脂肪酶催化的柞蚕丝织物接枝脂肪醇疏水整理

研究了柞丝绸用脂肪酶接枝脂肪醇提高其拒水性能。研究结果表明：柞丝绸接枝正十八醇的接枝率可以达到19.8%;接枝的脂肪醇级别越高,柞丝绸水接触角越大,沾水级别越高。水接触角从对照的55.8o提高到128.3o;沾水等级从0级提高到3级;接枝改性柞丝绸的透气性略有升高。洗涤试验表明：酯化接枝改性的柞丝绸具有良好的耐洗涤性能。因此,柞丝绸用脂肪酶接枝高级脂肪醇提高其拒水性能是一绿色、有效的整理方法。     

触角状环氧链结构对固定化脂肪酶(YCJ01)酶活的影响

为了改善酶的刚性固定化对酶蛋白构象的负面影响,以原子转移自由基聚合法（ATRP）合成了亲水性、柔性、触角状环氧化酶固定化载体PS-acyl-P（AM-co-GMA）,通过改变单体总量与引发剂量比例得到不同链长触角状环氧化酶固定化载体,并将其用于耐有机溶剂脂肪酶（YCJ01）的共价柔性固定化,重点考察了链长对固定化酶酶活的影响。结果表明,链长（增重率不超过3200%）越长,酶活越高。     

酶法改性环氧大豆油的流变学与摩擦学性能研究

为扩大大豆油的应用范围,采用脂肪酶Novozym435催化大豆油环氧化改性合成环氧大豆油,对环氧大豆油进行了红外光谱和核磁共振氢谱表征,并考察其流变学和摩擦学性能。结果表明：成功合成了环氧大豆油;相较于大豆油,环氧大豆油的黏度指数增大,黏温性能更优异;相较于矿物润滑油和大豆油,环氧大豆油的平均摩擦系数及磨损体积减少率分别为21.6%、17.0%和71.2%、64.9%,减摩抗磨性能显著提升。环氧大豆油可在摩擦表面形成致密、厚实的物理化学吸附膜,从而有效减摩抗磨,提高润滑性。