非水相中固定化脂肪酶催化性能的研究

以离子交换树脂为载体,用离子交换吸附法固定化脂肪酶,对影响固定化过程的各种因素进行了考察,确定了最优条件。得到的固定化酶催化合成月桂酸月桂醇酯,考查了在不同lgP值有机溶剂中的催化反应,异辛烷为最佳反应介质,最佳水含量为8％,最佳酸醇摩尔比为1：2,最适催化温度为55℃,得到的活化能Eα为19．00kJ／mol。     

Catalysis in Organic Solvents with Lipase Immobilized by Sol-Gel Technique

A series of immobilized lipases were obtained by sol-gel process, using silica prepolymers prepared from tetramethoxysilane, methyltrimethoxysilane, propyltrimethoxysilane and (3-aminopropyl)triethoxysilane. The activities of these biocatalysts were compared with the lipase adsorbed on poly(methylhydroxysiloxane) and encapsulated into a silicone rubber, lipase entrapped in nanoporous silica matrix and commercial sol-gel lipase. Model reactions were the esterification of stearic acid and Corey lactone bisalcohol (an intermediate of prostaglandin synthesis). The positive effect of hydrophobic-hydrophilic interface, created by the addition of organosilanes, on the activity of biocatalysts was partially reduced by decreasing specific surface of mesopores. Hydrophobic solvents increased the activity of the lipase entrapped in tetramethoxysilane–methyltrimethoxysilane prepolymer in the sequence acetone < toluene < benzene < decane < hexane. The activity of silicone rubber-encapsulated biocatalysts was proportional to polymer swelling in organic solvents (hexane > toluene > acetone).     

新型树枝状分子复合载体的合成及其对脂肪酶的固定化

新型树枝状分子复合载体的合成及其对脂肪酶的固定化;猪胰;树枝状分子二氧化硅复合载体     

纳米-微米复合孔泡沫陶瓷固定化脂肪酶

考察了泡沫陶瓷的孔径分布和表面性质对脂肪酶固定化的影响.研究表明,泡沫陶瓷的纳米孔孔径分布非常适合脂肪酶的固定化,对固定化酶的催化效率有决定性的影响.经1h的定化,泡沫陶瓷固定化酶的活性达商业化硅藻土固定化酶的1.33倍,体积活力为其2.63倍,蛋白载量为45.36mg/g陶瓷,比活为1215.39U/g,活力回收为41.2%.泡沫陶瓷固定化脂肪酶在有机相乙酸乙酯合成中表现优良,连续使用5次,每次反应3h,乙酸转化率均在93%左右.     

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。     

甲壳胺-硅胶复合载体的制备及其在脂肪酶固定化中的应用

固定化脂肪酶;甲壳胺-硅胶复合载体的制备及其在脂肪酶固定化中的应用     

Rapid Screening of Lipase Inhibitors in Scutellaria baicalensis by Using Porcine Pancreatic Lipase Immobilized on Magnetic Core–Shell Metal–Organic Frameworks

As for ligand fishing, the current immobilization approaches have some potential drawbacks such as the small protein loading capacity and difficult recycle process. The core–shell metal–organic frameworks composite (Fe₃O₄-COOH@UiO-66-NH₂), which exhibited both magnetic characteristics and large specific surface area, was herein fabricated and used as magnetic support for the covalent immobilization of porcine pancreatic lipase (PPL). The resultant composite Fe₃O₄-COOH@UiO-66-NH₂@PPL manifested a high loading capacity (247.8 mg/g) and relative activity recovery (101.5%). In addition, PPL exhibited enhanced tolerance to temperature and pH after immobilization. Then, the composite Fe₃O₄-COOH@UiO-66-NH₂@PPL was incubated with the extract of Scutellaria baicalensis to fish out the ligands. Eight lipase inhibitors were obtained and identified by UPLC-Q-TOF-MS/MS. The feasibility of the method was further confirmed through an in vitro inhibitory assay and molecular docking. The proposed ligand fishing technique based on Fe₃O₄-COOH@UiO-66-NH₂@PPL provided a feasible, selective, and effective platform for discovering enzyme inhibitors from natural products.     

固定化脂肪酶催化合成6,6'-海藻糖月桂酸酯

 以吸附在硅藻土上的假丝酵母（Candidasp．1619）脂肪酶为催\r\n化剂，在无溶剂体系中，研究了月桂酸与海藻糖的酯化反应．结果表明\r\n，反应温度为47℃，pH为7．0，月桂酸与海藻糖的摩尔比大于6时酯化\r\n反应的转化率较高．在3．2mmol（0．64g）月桂酸、0．2mmol（0．07\r\n6g，即53μl浓度为1．43g／ml的糖溶液）海藻糖和200mg固定化脂肪酶\r\n（1000U）组成的反应体系中，于47℃振荡反应36h，以海藻糖月桂酸双\r\n酯计算，酯化程度达95％以上．用有机溶剂提取的反应产物经薄层色谱\r\n提纯后进行13CNMR分析，结果表明，产物主要是6，6’－海藻糖月桂酸\r\n酯，其纯度为90％～95％．     

Activity and Enantioselectivity of Lipase Immobilized in CTAB Microemulsion-based Gels

Introduction The formation of gelatin-containing microemulsionbased gels(MBGs)was first described in1986[1,2]and the physical/structural characterization was carried out by a number of groups[3,4]with a variety of techniques including tracer diffusion,electrical conductivity,NMR,X-ray and small angle neutron scattering.     

介孔分子筛SBA-15的脂肪酶固定量分析测定

利用吸附法将假丝酵母脂肪酶(candida rugosa lipase,CRL)固定在介孔分子筛SBA-15上,对比了由单波长紫外分光光度法、双波长紫外分光光度法和二辛可宁酸法(bicinchoninic acid method, BCA)法测定的酶蛋白浓度及酶蛋白固定量.结果表明: SBA-15对紫外吸收有明显干扰,单波长紫外法测定结果远大于双波长紫外法和BCA法,双波长紫外法和BCA法测定结果较接近.利用BCA法测定了不同浓度CRL在介孔分子筛上的固定量,考察了固定化酶的泄漏量.在编号分别为Lu001和LLSD1的介孔分子筛SBA-15上的载酶量分别为16.6和114.12 mg/g.在缓冲溶液中SBA-15固定化酶的泄漏率只约为0.5%,可作为良好的酶固定化载体.     

Enhancement of the Photochromic Performance of Spirooxazine in Sol-Gel Derived Organic-Inorganic Hybrid Matrices by Additives

Organic-inorganic hybrid materials synthesized via sol-gel processing are excellent solid matrices for photochromic dyes like spirooxazine, and the photochromic performance can be further enhanced by introducing suitable additives. This work describes the effects of additives on the photochromic intensity (ΔA ₀), decolouration rate (k) and photostability of spirooxazine in sol-gel derived organic-inorganic hybrid matrices. They include fluoroalkylsilane (FAS), bisphenolA (BPA) and methyl-imidazole (MI). FAS enhances both ΔA ₀ and photostability, but has little effect onk. The higher the content of BPA, the higher the ΔA ₀ and the higher the photostability, but the lower thek. The effect of MI on ΔA ₀ andk is not so considerable as that of BPA and is a little complicated, while significant improvement in photostability is achieved by the addition of MI at an optimum concentration with the presence of BPA at a higher content.     

混合溶剂系统中固定化脂肪酶对酮基布洛芬的催化酯化反应

以硅藻土吸附的脂肪酶为催化剂,对外消旋酮基布洛芬[2-(3-苯甲酰苯基)丙酸]进行对映选择性酯化反应;考察了不同的脂肪酶制剂,固定化时所加缓冲液的体积与pH值,酰基受体(醇)的种类以及混合溶剂系统的组成等因素对酶活性的影响.结果表明,在所考察的7种脂肪酶中,以LipaseOF的酪化活性最高;用硅藻土吸附固定化酶时,缓冲溶液的最适pH为7.0左右,每克酶粉加1.0mL缓冲溶液为最佳;固定化酶催化酯化的活性比游离的脂肪酶高.在酮基布洛芬与不同酰基受体(醇)的酶促酯化反应中,以丙醇的反应速度为最快.在由一种主溶剂与一种助溶剂组成的混合溶剂系统中,酶促酯化的速度要比在单一的主溶剂或助溶剂系统中快.当以1gP值较大的环己烷或异辛烷等为主溶剂,甲苯为助溶剂时,脂肪酶催化酮基布洛芬酯化反应的活性最高.     

溶胶-凝胶法制备有机/无机杂化材料工艺及其应用

介绍了溶胶–凝胶法制备有机/无机杂化材料的原理和基本过程,杂化材料的制备方法及对材料性能的影响,概述了杂化材料在结构材料、光学材料及其它材料中的应用研究。     

有机相中蚕丝固定化脂肪酶催化酯化反应性能研究

研究了有机相中蚕丝固定化脂肪酶催化酶化反应的催化活性。考究了有机溶剂,底物、反应温度,ＰＨ值和体系含水量等因素对固定化脂肪酶催化活性的影响。结果表明,以异辛烷为有机溶剂,在反应温度为５０℃ＰＨ值为７．４时,酶催化活性最好。     

聚乙烯亚胺改性聚丙烯腈中空膜固定脂肪酶

聚丙烯腈是富腈基的高分子聚合物,易修饰改性,广泛应用于膜分离应用.我们以聚丙烯腈中空膜为载体,采用化学法交联聚乙烯亚胺并固定脂肪酶,固定过程中引入海藻酸钠,用CaCl_2进行后处理,得到固定化脂肪酶PAN-PEI-SA/E-CaCl_2载酶量为31.70(mg enzyme)/(g support),酶活为50.20 U/(g support),15次重复使用可保留58.77%的酶活,与游离酶相比耐酸性和耐温性有所提高,相同条件下与Nov 435相比,酶活更高,这表明最终得到的固定化脂肪酶有良好的工业应用前景.     

Synthesis of Cysteine-Functionalized Silver Nanoparticles Using Green Tea Extract with Application for Lipase Immobilization

The application of cysteine-capped silver nanoparticles synthesized using green tea as the reducing agent to immobilize lipase has been reported in the present work. The reducing property of green tea is due to the presence of polyphenolic compounds in its extract which are not oxidized at ambient atmospheric conditions and hence is a suitable reducing agent for green synthesis of nanoparticles. Cysteine-capped silver nanoparticles were synthesized under alkaline conditions by reducing the silver salt by green tea extract in the presence of cystine. Various parameters such as the cystine concentration, pH, temperature, and amount of reducing agent were standardized and their effect on the synthesis process has been initially evaluated by surface plasmon resonance peak analysis. Furthermore, the synthesized nanoparticles were also characterized using X-ray diffraction, Fourier transform infrared spectroscopy, and transmission electron microscopy. The particle size analysis revealed the average size of the particles to be around 20?nm. The glutaraldehyde-deactivated amino group on cysteine-capped nanoparticles was used to immobilize lipase on its surface. Both crude and immobilized lipases were checked for activity and protein content under standard assay conditions and their activity was found to be 37.7 and 24.9?U?mL^?1, respectively. The lipase nanoparticle bioconjugates exhibited a good shelf life of 60 days with a marginal decrease in activity. The bioconjugates showed 15% loss in its initial activity at the end of five reusability cycles. This immobilized reusable system has the potential to be utilized for various applications pertaining to the exploitation of lipase in various industries.     

Catalytic Characteristics of Lipase Immobilized in CTAB/TX-100 MBG

CTAB/TX-100 microemulsion-based gel(MBG), which could be soaked in aqueous solution for a long time without mechanical strength and shape changes, was successfully prepared. Lipase immobilized in this gel had a higher activity than that in CTAB MBG when catalyzed the esterification reaction between n-hexanoic acid and n-octanol and its enantioselectivity was slightly higher when catalyzed the stereoselective esterification reaction between racemic ibuprofen and n-octanol. In aqueous solution, lipase in CTAB...     

Auto-Organisation in Silica-Based Organic-Inorganic Gels Obtained by Sol-Gel Process

A new type of hybrid material is prepared through sol-gel processing by the polycondensation of (MeO)₃Si–R–Si(OMe)₃ units containing a rigid organic anisotropic group R. The hybrid covalently bounded organic-inorganic gel shows an unexpected structural birefringence n (n = 2 × 10^–3). This birefringence is induced by a strain field anisotropy during gel aging as revealed by simultaneous dynamic light scattering and birefringence measurements. To better address the role of the strain field, we use a free interface sol-gel/air to control strain anisotropy and to measure the gel optical axis and its birefringence. We find that the birefringence is associated to a long-range orientational order of organic moieties induced by the strain anisotropy during the gel aging.     

固定化脂肪酶催化碳酸二甲酯制备生物柴油的研究

以介孔硅材料(MPS)为载体将脂肪酶固定化,以碳酸二甲酯为酰基受体,对固定化酶催化碳酸二甲酯进行了反应路径(原料油)、反应条件(反应温度、碳酸二甲酯的用量、加水量)的优化,在最佳的条件下对实验过程中所用的固定化酶进行重复使用性的考察.实验结果表明,不同种油与碳酸二甲酯反应在同定化酶的催化下制备生物柴油的产率以麻疯树油为最高,最佳反应条件是碳酸二甲酯的浓度为16mL/g、不加水,在50℃下反应24h,生物柴油得率达81.6％.     

Synthesis of Active Porous Silica Gel Matrices and Investigation of Their Properties

Adsorption-structural and mechanical properties and heat treatment behaviour of alkoxy-derived hydrogels prepared by acid-catalyzed reaction of tetraethoxysilane have been investigated. Conditions for synthesizing xerogels as transparent, active mesoporous matrices have been found. The relation between the amount of immobilized optical active components and the textural features of matrices have been investigated.