聚ε-己内酯的微米硅球固定化猪胰脂肪酶促合成

本文采用微米硅球固定化猪胰脂肪酶为催化剂合成聚ε-己内酯, 以期获得具有较高分子量、 良好生物相容性和使用安全性的生物可降解医用高分子材料.     

Enzymatic ring-opening polymerization of 2,2-dimethyltrimethylene carbonate catalyzed by PPL immobilized on silica nanoparticles

Silica nanoparticles were first used as the carrier for the porcine pancreas lipase (PPL) immobilization. The result of transmission electron microscopy (TEM) showed that the immobilized lipase was still in nanosize after enzyme immobilization. The ring-opening polymerization of 2,2-dimethyltrimethylene carbonate (DTC) catalyzed by this immobilized PPL (IMPPL) was explored. ¹H NMR spectra suggested no evidence of decarboxylation during propagation. Influences of IMPPL concentration and reaction temperature on the molecular weight and yield of poly(DTC) were studied. The recovery and reuse of IMPPL for the ring-opening polymerization of DTC was also investigated. The recycling IMPPL showed even higher catalytic activity and a higher molecular weight of polycarbonate could be achieved.     

Synthesis of functional polycarbonates by lipase-catalyzed ring-opening polymerization

Poly(5-benzyloxy-trimethylene carbonate) (PBTMC), a new functional polycarbonate was synthesized by enzymatic ring-opening polymerization in bulk at 150°C using Porcine pancreas lipase (PPL) or Candida rugosa lipase (CL) as catalyst. Influences of different polymerization conditions such as the source of enzyme, enzyme concentration and polymerization time on the molecular weight and yield were studied. The results showed that PPL exhibited higher activity than CL. Both higher molecular weight(Mn, 18953) and yield(98%) could be obtained by the use of PPL as catalyst. ¹H NMR spectrum showed no decarboxylation occurrence during the ring-opening polymerization.     

大环碳酸酯的Novozym-435酶促开环聚合

研究了14元环的碳酸丁二酯二聚体在固定化脂肪酶Novozym-435催化下的开环聚合反应制备聚碳酸丁二酯.聚合在常压,75℃的甲苯溶液中进行,反应条件温和.详细探讨了反应条件诸如单体浓度,酶浓度对于聚合的影响.结果显示Novozym-435具有与异辛酸亚锡可比拟的高催化活性,同时可以回收重复使用.聚合动力学研究表明碳酸丁二酯的酶促甲苯溶液开环聚合和环状内酯的酶促甲苯溶液聚合有所不同,没有表现出活性聚合的特征.     

一种含有氯乙氧功能基团的环状磷酸酯的酶促开环聚合

作为生物体内广泛存在的一类高分子——聚磷酸酯以其良好的生物相容性、生物可降解性及低毒性,一直受到人们的广泛关注．尤其是一些含有功能基团的聚磷酸酯,在基因转染、药物控制释放和组织工程等领域具有很好的应用前景．聚磷酸酯可通过金属催化剂开环聚合制备而成,但残留的微量金属催化剂可能会给材料带来潜在毒性．     

环状磷酸酯的酶促开环聚合

研究了环状磷酸酯的酶促开环聚合反应,讨论了环状磷酸酯取代烷基对于酶促开环聚合及相应聚磷酸酯性能的影响,发现烷基取代基长度对于聚合度没有明显影响; 但随着环状磷酸酯的取代基长度增加,产率随之降低,聚磷酸酯的亲脂性增强. 猪胰脂肪酶和假丝酵母皱褶酶显示出比碱性磷脂酶更高的活性.     

Lipase‐catalyzed ring‐opening polymerization of 6(S)‐methyl‐morpholine‐2,5‐dione

Lipase‐catalyzed ring‐opening bulk polymerizations of 6(S)‐methyl‐morpholine‐2,5‐dione (MMD) were investigated. Selected commercial lipases were screened as catalysts for MMD polymerization at 100 °C. Polymerizations catalyzed with 10 wt % porcine pancreatic lipase type II crude (PPL), lipase from Pseudomonas cepacia, and lipase type VII from Candida rugosa resulted in MMD conversions of about 75% in 3 days and in molecular weights ranging from 8200 to 12,100. Poly(6‐methyl‐morpholine‐2,5‐dione) [poly(MMD)] had a carboxylic acid group at one end and a hydroxyl group at the other end. However, lipase from Mucor javanicus showed lower catalytic activity for the polymerization. During the polymerization, racemization of the lactate residue took place. PPL was selected for further studies. The rate of polymerization increased with increasing PPL concentration under otherwise identical conditions. When the PPL concentration was 5 or 10 wt % with respect to MMD, a conversion of about 70% was reached after 6 days or 1 day, respectively, whereas for a PPL concentration of 1 wt %, the conversion was less than 20% even after 6 days. High concentrations of PPL (10 wt %) resulted in high number‐average molecular weights (<3 days); with a lower concentration of PPL, lower molecular weight poly(MMD) was obtained. The concentration of water was an important factor that controlled not only the conversion but also the molecular weight. With increasing water content, enhanced polymerization rates were achieved, whereas the molecular weight of poly(MMD) decreased. © 2005 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 43: 3030–3039, 2005     

Preparation method of porous polymer materials by radiation technique and its application

The radiation polymerization of hydrophilic hydroxyethyl methacrylate monomer solution at temperatures below 0 °C leads to the formation of a porous structure in the polymers. The melting peak of the eutectic water–monomer composition at the eutectic point (above ?24 °C) could be distingushed and a glass transition temperature was observed at ?96 °C. The porous structure was developed after melting small ice pieces in the polymers after polymerization. The porous structure formed above 0 °C contained discontinuous pores and that formed below 0 °C had continuous pores leading to reticular structure. In a mixture of water –dioxane –monomer, the pore diameter decreased with increasing monomer concentration. Replacing dioxane with decane led to a maximum pore diameter at 70% monomer concentration. The pore diameter in 70% monomer concentration using water and dioxane was 1～4 µm, maximum activity in immobilized enzyme tablets was observed at this diameter. The porous structure was also varied by controlling the polymerization temperature. The durability of the immobilized enzyme tablets was demonstrated by the retention of high enzyme activities after repeated batch enzyme reactions. Copyright © 2001 John Wiley & Sons, Ltd.     

单分散磁性纳米粒子固定化猪胰脂肪酶的研究

借助溶热法制备了一种亲水及生物相容良好的Fe3O4磁性纳米粒子,用γ-氨丙基三乙氧基硅烷直接对所得磁性粒子表面改性,然后用戊二醛偶联法制得了固定化猪胰脂肪酶.表征研究显示,所得磁性粒子粒径约200 nm,具有良好的单分散性和磁响应性.考察了戊二醛浓度、给酶量和反应时间对脂肪酶固定化过程的影响,并通过游离酶与固定化酶的比...     

聚脲多孔材料的简单制备及在酶固定和手性拆分中的应用

以甲苯二异氰酸酯(TDI)为单体,在水与丙酮混合溶剂中通过沉淀聚合一步法制备了富含胺基的聚脲多孔材料(PPU),通过扫描电镜和压汞法对其表面形貌和孔结构进行了表征.PPU经戊二醛(GA)活化后用于荧光假单胞菌脂肪酶(PFL)的固定,考察了GA活化过程中GA浓度对酶固定量及固定酶活性的影响.结果表明,PPU是一种粒子尺寸分布在30~50μm范围的形状不规则的多孔粒子,孔径在2 nm~100μm之间呈连续分布.在pH=8.0的缓冲溶液中用0.17 mol/L的GA对PPU进行改性,将改性后的PPU用于PFL的固定,当酶溶液浓度为2.56 mg/m L时,得到酶的最大固定量为95.2 mg/g,固定酶的活性为375 U/mg,相对活性为76%.将此固定酶作为催化剂,用于1-苯乙醇外消旋化合物的手性拆分,并与游离酶催化的结果相比较.结果表明,固定酶的反应活性和立体选择性都明显优于游离酶.通过沉淀聚合制备的聚脲多孔材料在酶固定及手性分子拆分方面具有应用前景.     

Immobilization of β-Galactosidase onto Magnetic Beads

A study of the cross-linking of β-galactosidase on magnetic beads is reported here. The magnetic beads were prepared from artemisia seed gum, chitosan, and magnetic fluid in the presence of a cross-linking regent (i.e., glutaraldehyde). The reactive aldehyde groups of the magnetic beads allowed the reaction of the amino groups of the enzymes. The animated magnetic beads were used for the covalent immobilization of β-galactosidase. The effect of various preparation conditions on the activity of the immobilized β-galactosidase, such as immobilizing time, amount of enzyme, and the concentration of glutaraldehyde, were investigated. The influence of pH and temperature on the activity and the stability of the enzyme, both free and immobilized, have been studied. And o-nitrophenyl-β-d-galactopyranoside (ONPG) was chosen as a substrate. The β-galactosidase immobilized on the magnetic beads resulted in an increase in enzyme stability. Optimum operational temperature for immobilized enzyme was 10 °C higher than that of free enzyme and was significantly broader.     

Samarium enolate on crosslinked polystyrene beads. III. Anionic initiator for well‐defined synthesis of poly(hydroxyethyl methacrylate) on solid support

A solid‐supported samarium enolate successfully initiated the polymerization of 2‐(trimethylsilyloxy)ethyl methacrylate (TMS‐HEMA) through the living anionic process. In addition, the silyl group was readily removed by treatment of the beads with a weak acid to afford the corresponding well‐defined poly(methacrylate) having a hydroxyethyl group in the side chain (PHEMA). The hydroxyl group of the immobilized PHEMA on the beads was successfully acetylated to give poly(2‐acetoxyethyl methacrylate), which could be quantitatively isolated from the beads by trifluoroacetic acid treatment. Moreover, the hydroxyl group of the immobilized PHEMA could be utilized as an initiator for acid promoted ring opening polymerization of lactone to yield the corresponding graft copolymer. In this method, the residual and excess reagents could be removed by filtration, which demonstrated the applicability of the present technique to a novel method for construction of functional polymers. © 2004 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 42: 4417–4423, 2004     

Ethylene (Co)Polymerization with Metallocene Catalysts Encapsulated in Gel‐Type Poly(styrene‐co‐divinylbenzene) Beads

Gel‐type poly(styrene‐co‐divinylbenzene) beads (PS bead) were used as a carrier to encapsulate metallocene catalysts through a simple swelling‐shrinking procedure. The catalytic species were homogeneously distributed in the PS bead particle. The catalyst exhibited high and stable ethylene polymerization and ethylene/1‐hexene copolymerization activity affording uniform spherical polymer particles (1 mm). Polymerization rate profiles exhibited slow initiation and stable increase in polymerization activity with time.     

Ascorbic acid determination in biological fluids using ascorbate oxidase immobilized on alkylamine glass beads in a flow injection potentiometric system

A flow injection method was developed aimed at the determination of ascorbic acid in biological fluids, particularly fruit juices. The enzyme ascorbic oxidoreductase (EC 1.10.3.3), extracted fromCucurbita maxima, was immobilized onto alkylamine glass beads using glutaral-dehyde as a bifunctional agent. The ascorbic acid concentration was related to oxygen saturation. Fall in oxygen concentration, as a result of ascorbic acid oxidation, was detected by a low cost, homemade oxygen electrode. The calibration graph was linear over the range 0.05 to 3.00 mM (RSD 1%), the maximum number of samples that could be analysed was 90/h. The immobilized enzyme retained its initial activity for 2 mo with more than 600 assays.     

Atom transfer radical polymerization of methyl methacrylate catalyzed by ion exchange resin immobilized Co(II) hybrid catalyst

Ion exchange resin immobilized Co(II) catalyst with a small amount of soluble CuCl₂/Me₆TREN catalyst was successfully applied to atom transfer radical polymerization (ATRP) of methyl methacrylate (MMA) in DMF. Using this catalyst, a high conversion of MMA (>90%) was achieved. And poly(methyl methacrylate) (PMMA) with predicted molecular weight and narrow molecular weight distribution (M_w/M_n = 1.09–1.42) was obtained. The immobilized catalyst can be easily separated from the polymerization system by simple centrifugation after polymerization, resulting in the concentration of transition metal residues in polymer product was as low as 10 ppm. Both main catalytic activity and good controllability over the polymerization were retained by the recycled catalyst without any regeneration process. © 2008 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 46: 1416–1426, 2008     

含环氧基亲水性固定化青霉素酰化酶共聚载体的合成与性能研究

环氧基团可以在温和条件下与酶分子的氨基反应使其固定于载体表面.选用含有活性环氧基团的甲基丙烯酸缩水甘油酯(GMA)和亲水性的N-乙烯吡咯烷酮(NVP)两种单体,以N,N′-亚甲基双丙烯酰胺(MBAA)为交联剂,甲醇水溶液作致孔剂,液体石蜡为主介质,通过反相悬浮聚合技术成功地合成了亲水性大孔GMA-NVP-MBAA三元共聚物载体(GNM).通过调节交联剂的用量和单体NVP与GMA的比例,可以调节载体的孔径、比表面积及在水中的溶胀性能.将巨大芽孢杆菌青霉素酰化酶共价偶联于平均孔径为16.5nm、表面环氧基含量为0.906mmol/g的GNM共聚物载体,制成固定化酰化酶,其表观活性高达625U/g,水解青霉素G钾盐的最适宜温度为50℃,pH值为8.0.固定化酶在4℃保存40d,活性保持不变.经3次使用后,活性达到稳定值(601U/g左右),再经12次使用,活性几乎保持不变.     

猪胰脂肪酶的固定化及其在有机相中催化丁酸甲酯和正丁醇的酯交换反应研究

考察了大孔苯乙烯－二乙烯苯交联共聚物的交联度、致孔剂量及胺化试剂对载体固定化猪胰脂肪酶的影响。选择出一种最佳载体对猪胰脂肪酶进行固定化，对比了自由酶和固定化酶在有机相中催化丁酸甲酯和正丁醇的酯交换反应。结果表明，酶经固定化后催化反应活力比自由酶提高近１倍。     

Concanavalin a immobilized affinity adsorbents for reversible use in yeast invertase adsorption

Concanavalin A (Con A) immobilized poly(2-hydroxyethyl methacrylate) (PHEMA) beads were investigated for specific adsorption of yeast invertase from aqueous solutions. PHEMA beads were prepared by a suspension polymerization technique with an average size of 150-200 microm, and activated by epichlorohydrin. Con A was then immobilized by covalent binding onto these beads. The maximum Con A immobilization was found to be 10 mg/g. The invertase-loading capability of the PHEMA/Con A beads was 107 mg/g. The maximum invertase adsorption capacity on the PHEMA/Con A adsorbents was observed at pH 5.0. The values of the Michaelis constant K(m) of invertase were significantly larger upon adsorption, indicating decreased affinity by the enzyme for its substrate, whereas V(max) was smaller for the adsorbed invertase. Adsorption improved the pH stability of the enzyme as well as its temperature stability. Thermal stability was found to increase with adsorption. The adsorbed enzyme activity was found to be quite stable in repeated experiments. Storage stability of adsorbed invertase.     

Optimum conditions for preparing micron-sized PMMA beads in the dispersion polymerization using PVA

The optimum conditions for preparing micron-sized monodisperse polymethylmethacrylate (PMMA) beads by dispersion polymerization in a methanol/water mixture were proposed. PMMA forming microspheres having an average molecular weight of 55,300 g/mol, 2.6 μm weight-average diameter, with a 5.3% coefficient of variation and 91% conversion, were successfully obtained in the presence of 15 wt.% of polyvinylalcohol (PVA), 100/50 (g/g) of MeOH/water mixtures at 70°C; the reaction lasted for 8 h. Compared to dispersion polymerization using polyvinylpirrolydone, PVA proved to be an extremely stable steric stabilizer in the dispersion polymerization of methylmethacrylate.     

Effect of temperature cycling on the activity and productivity of immobilized β-galactosidase in a thermally reversible hydrogel bead reactor

The enzyme beta-galactosidase has been immobilized within thermally reversible hydrogel beads that exhibit LCST (lower critical solution temperature) behavior. The hydrogel beads containing the immobilized enzymes swell and expand below the LCST and deswell and shrink above the LCST. This behavior is reversible. The enzyme was physically entrapped in a crosslinked hydrogel of a copolymer of N-isopropylacrylamide (NIPAAm) and acrylamide (AAm), and formed as beads in an inverse suspension polymerization. The beads were placed in a packed bed column reactor which was operated in a continuous, single pass mode, either isothermally at 30 or 35 degrees C, or with temperature cycling between 30 and 35 degrees C. The thermal cycling significantly enhanced overall reactor enzyme activity relative to isothermal operation at either the higher or lower temperature. It is postulated that mass transfer rates within the hydrogel beads are greatly enhanced by the movement of water in and out of the beads during the expansion or collapse of the polymer chain network as temperature is cycled.