天然矿物埃洛石为载体固定α-淀粉酶的工艺和性能

以天然纳米材料埃洛石为载体,通过物理吸附法对α-淀粉酶进行固定.利用红外光谱、扫描电镜、透射电镜等对埃洛石的结构和形貌特征进行测试与表征,同时对埃洛石纳米管固定化α-淀粉酶的条件及固定化酶的酶学性能进行了研究,并与游离酶进行了比较.结果表明:这种具有管状结构的埃洛石硅酸盐矿物是理想的酶载体,酶的固定化效率平均达到37.38%;所得的固定化α-淀粉酶4℃下保存15d后,酶活力仍保持90%以上;固定化α-淀粉酶的热稳定性也明显优于游离酶,连续7批次操作后仍保持56.2%的酶活力.     

α-淀粉酶在MCM-41介孔分子筛上的固定化研究

采用浸渍法将α-淀粉酶固定在介孔分子筛MCM-41上。考察了吸附时间、给酶量和pH对α-淀粉酶固定化性能的影响,并对固定化酶的活性、稳定性和载体结构等进行了研究。结果表明,在固定化时间为11 h,给酶量为70 mg.g-1,pH=5.9的条件下,固定化酶活性回收率可达48%。与游离酶相比,固定化酶的耐热能力增强,温度达到70℃时,固定化酶相对活性可达到75%,而游离酶只有14%;在pH=3.3～8.0的内,固定化酶相对活性为62%～100%,而游离酶的相对活性为5%～100%,固定化酶具有更宽的pH适应性;此外,固定化酶储存稳定性明显增强,并具有一定的可重复操作性,且固定后载体仍然保持了良好的介孔结构。     

Bioaffinity Adsorption Behavior of Several Enzymes onto β-Cyclodextrin-Chitosan

Abstract

In this study, specific and nonspecific adsorption and desorption of α-amylase, lactase, and amyloglucosidase (AMG) enzymes onto the β-cyclodextrin (CD)-chitosan system were investigated. α-Amylase and lactase enzymes were adsorbed onto the chitosan-only and chitosan-spacer gels, but very little adsorption was observed in the case of the chitosan gels with CD. Furthermore, AMG enzyme showed a high degree of interaction with CD immobilized on chitosan. Therefore, the interaction between the AMG and CD molecules can be explained by the fact that there is true biospecific adsorption. Desorption experiments also confirm this theory because the elution of AMG was much more difficult than the elution of lactase and α-amylase from the CD matrix.     

Acid activated montmorillonite: an efficient immobilization support for improving reusability,storage stability and operational stability of enzymes

Three enzymes, α-amylase, glucoamylase and invertase, were immobilized on acid activated montmorillonite K 10 via two independent techniques, adsorption and covalent binding. The immobilized enzymes were characterized by XRD, N₂ adsorption measurements and ²⁷Al MAS-NMR spectroscopy. The XRD patterns showed that all enzymes were intercalated into the clay inter-layer space. The entire protein backbone was situated at the periphery of the clay matrix. Intercalation occurred through the side chains of the amino acid residues. A decrease in surface area and pore volume upon immobilization supported this observation. The extent of intercalation was greater for the covalently bound systems. NMR data showed that tetrahedral Al species were involved during enzyme adsorption whereas octahedral Al was involved during covalent binding. The immobilized enzymes demonstrated enhanced storage stability. While the free enzymes lost all activity within a period of 10 days, the immobilized forms retained appreciable activity even after 30 days of storage. Reusability also improved upon immobilization. Here again, covalently bound enzymes exhibited better characteristics than their adsorbed counterparts. The immobilized enzymes could be successfully used continuously in the packed bed reactor for about 96 hours without much loss in activity. Immobilized glucoamylase demonstrated the best results.     

A novel method for continuous production of cyclodextrins using an immobilized enzyme system

Selected specific α-amylase and glucoamylase enzymes were immobilized and combined with immobilized cyclodextrin glycosyltransferase to continuously produce cyclodextrin (CD). The liquefied starch was cyclized to CD by ICGTase and then treated by immobilized α-amylase and glucoamylase to contain CD and glucose only. β-CD and soluble CD (α- and γ-CD) in the final CD solution could be easily separated. The pH-activity curve, temperature-activity curve, batch re-usability and continuous operation stability of immobilized enzymes were discussed. The continuous production of CD by an immobilized enzyme system was also reported. The optimal conditions for using immobilized α-amylase and glucoamylase simultaneously were 40°C and pH 4.5 adjusted by 1 mol dm^?3 HCl/NaOH. A 70% yield of CD could be obtained from 1% (w/v) of liquefied starch under continuous operation at 0.055 h^?1 space velocity, and almost all the oligosaccharides (β 98%) were converted to glucose. In this study, the separation of α-CD, γ-CD and glucose, using organic solvent from the final product after precipitating β-CD, was also investigated.     

Characterization of surface‐modified poly(ethylene terephthalate) fibres by inverse gas chromatography

The effect of surface cleanliness on the alkaline hydrolysis of poly(ethylene terephthalate) fibres was investigated using inverse gas chromatography (IGC) in conjunction with mass loss measurements and electron microscopy. The sizing agent was removed from the fibre surface by two methods: soxhlet cleaning in acetone and washing in an aqueous solution of a non‐ionic detergent. Alkaline hydrolysis was carried out using two concentrations of aqueous sodium hydroxide, 1% and 10% by mass. The measurement of the specific retention volume of undecane and the heat of adsorption using IGC indicated that the acetone cleaned samples were essentially surface contaminant free, while partial contamination of the surface by the sizing agent remained in the detergent cleaned samples. The presence of sizing agent significantly altered the degree of hydrolysis and the surface topography. The increasing values of the heat of adsorption indicated that significant surface hydrolysis increased the surface crystallinity. © 2000 Society of Chemical Industry     

Enzymes immobilized on montmorillonite K 10: Effect of adsorption and grafting on the surface properties and the enzyme activity

The enzymes α-amylase, glucoamylase and invertase were immobilized onto acid activated montmorillonite (K 10) via two independent techniques — adsorption and grafting using glutaraldehyde as spacer. Samples were characterized by XRD, NMR, SEM and BET N₂ adsorption measurements. Activity of the immobilized enzymes was tested in batch and bed reactor and their kinetic parameters were determined. XRD shows an increase of the basal spacing from 1 nm for montmorillonite to about 2.5 nm for immobilized invertase. It indicates that side chains of the enzymes have penetrated between the layers, in case of grafted enzymes, to a higher extent than for the adsorbed enzymes. ²⁷Al-MAS NMR reveals the interaction of octahedral and tetrahedral Al species with the enzyme. Adsorption affects the tetrahedral Al resonance while grafting involves the octahedral sites only. Surface area measurements illustrate that enzyme grafting reduces the porosity than enzyme adsorption. It is proposed that the enzymes are situated at the periphery of the clay mineral particles whereas the side chains of different amino acid residues penetrate between the layers. The immobilized enzymes show substantial activity. The higher activity in the packed-bed reactor indicates a better mass transport. Loss of activity in the batch reactor is due to diffusional mass transfer resistances.     

Cu(II) ions removal from wastewater using starch nanoparticles (SNPs): An eco-sustainable approach

The complex structured starch particles were reduced to the nanoscale size range through hydrolysis utilizing low concentration acid assisted by ultrasound irradiation. The synthesized starch nanoparticles (SNPs) were characterized by transmission electron microscopy (TEM), Fourier-transform infrared (FTIR), and X-ray diffraction (XRD) techniques. The synthesized SNPs possessed surface activated entities, as many cationic functional groups were confirmed through the FTIR spectrum. Also, these SNPs were effectively utilized to separate heavy Cu metal ions from the synthetic ion solution. The SNPs were characterized using field emission scanning electron microscope (FESEM), X-ray photoelectron spectroscopy (XPS), and Brunauer–Emmett–Teller (BET) analysis for the surface modification after the adsorption process. The weak electrostatic interaction between the SNP surface and Cu ion was confirmed by the XPS spectrum and energy-dispersive X-ray spectroscopy. The maximum efficiency of Cu ions removal was about 93% at an optimal pH 5 and 25 mg/ml dosage. The adsorption equilibrium was obtained in 60 min. The nitrogen isotherm BET analysis of SNPs after adsorption shows a higher specific surface area of 18.552 m²/g, attributed to the interaction and presence of Cu ions on the SNP surface. The process feasibility was validated by the Langmuir isotherm model. The process exhibits pseudo-second-order adsorption kinetics and follows the Langmuir isotherm. The R_L predicted by the Langmuir isotherm mechanism is 0.017, implying favourable adsorption. The process is reproducible and allows for the separation of heavy metal ions from the wastewater through biosorption effectively.     

Optimizing Detergent Formulation with Enzymes

Enzymes have been widely used in modern detergents. However, few studies deal with research on enzyme dosage as well as its relationship with other ingredients within detergent products. This paper compares the washing performance of Chinese phosphate and non-phosphate detergent before and after adding different enzymes and additives on the various types of swatches. The results confirmed that the detergent washing performance with enzymes added improved significantly for the specific stain soil. And within a certain range, the detergent washing performance and enzyme content have positive correlation. Furthermore there is synergy of soil removable ability between protease and amylase. Meanwhile the citric acid and various additives with enzymes can effectively compensate for insufficient of non-phosphate detergent.     

乙基纤维素固定化α-淀粉酶的研究

对乙基纤维素固定化α-淀粉酶进行了研究,优化了α-淀粉酶的固定化工艺条件,并比较了游离酶和固定化酶的特性。结果表明,在α-淀粉酶浓度为4g·L-1、乙基纤维素质量分数为0.50%、4℃的条件下,固定化α-淀粉酶的重复操作稳定性最好;固定化α-淀粉酶的最适反应pH值为7.0、最适反应温度为60℃,具有良好的热稳定性、重复使用性和储存稳定性;该固定化方法操作简便,减少了酶变性的可能,最大程度保留了酶的活力。     

Magnetic porous carbon microspheres synthesized by simultaneous activation and magnetization for removing methylene blue

Magnetic porous carbon microspheres (MPCMs) based on Fe₃O₄-encapsulating carbon composites for removing methylene blue (MB) in aqueous solutions were synthesized by simultaneous activation and magnetization. A series of MPCMs were prepared by combining hydrothermal and annealing treatment with α-Fe₂O₃ nanoparticles as iron source, glucose as carbon source and ZnCl₂ as porogen. The phase structure, specific surface area, porosity, thermostability, magnetic property, as well as morphology of as-prepared MPCMs were verified by X-ray diffraction, Brunauer–Emmeltt–Teller surface area analysis, thermogravimetric analysis, vibrating sample magnetometry, field emission scanning electron microscopy and high resolution transmission electron microscopy. The results indicate that the maximum specific surface area of MPCMs is up to 480.32 m²/g when the mass ratio of ZnCl₂/glucose is 0.25, which is designated as MPCMs-0.25. The saturation magnetism of MPCMs-0.25 is 30.16 emu/g. Adsorption properties of MPCMs were detected by using MPCMs-0.25 as adsorbent to remove MB from aqueous solution. The outcomes suggest that the adsorption reaches equilibrium within 35 min and physical adsorption is involved in the whole adsorption processes. The results of adsorption isotherm reveal that the adsorption process might include monolayer and porous adsorption, meanwhile, various adsorption sites exist on the surfaces of MPCMs-0.25. The reusability and stability of MPCMs-0.25 were also confirmed by five adsorption–desorption cycle experiments.     

Synthesis of Octa(maleimidophenyl) silsesquioxane–SiO2/TiO2 Hybrid Nanocomposites: Adsorption Behavior for the Removal of an Organic Methylene Blue Dye and Antimicrobial Activity against Pathogens

In this study, a sol–gel process was used to prepare hybrid nanocomposite consisting of octa(maleimidophenyl) silsesquioxane-silica/titania (maleimide–POSS (polyhedral oligomeric silsesquioxanes)–SiO₂/TiO₂) to use in methylene blue (MB) adsorption and as an antibacterial agent. The structure, surface, and morphological characteristics were confirmed through Fourier transform infrared spectroscopy, X-ray diffraction, X-ray photoelectron spectroscopy, and transmission electron microscopy. The optical and thermal stabilities were studied by conducting UV–visible and thermogravimetric analysis–differential scanning electron calorimetry analysis. The experimental results showed a maximum dye adsorption capacity of 92% achieved using 0.5?g of hybrid nanocomposite after 2.5?h at pH 9. We also investigated the effect that the adsorbent dosage, pH, and contact time had on the removal efficiency of the MB dye in aqueous solution. The recycling experiment showed a good adsorption capacity of the MB dye, even after five repeated cycles. Furthermore, the hybrid nanocomposite was tested against pathogenic bacteria, such as Bacillus cereus, Lactobacillus, Escherichia coli, and Pseudomonas aeruginosa. The nanocomposite was observed to be highly sensitive to E. coli, B. cereus, and P. aeruginosa, as confirmed by the size of the zone inhibition.     

酶在洗涤剂中的应用现状及展望

论述了洗涤剂用酶的发展史和常用4种酶(特别是纤维素酶)的特性、去污机理、功能和应用现状。指出了近年来洗涤剂用酶的动向：开发具有耐氧化性、低温下活性高、对洗涤剂和洗涤环境适应性好的变异蛋白酶已商品化;一些反应性更好,对油污吸附性更强的改良脂肪酶已出现;一类在高碱性、低温介质中能充分保持活性的淀粉酶正在积极开发中;一类纤维素复合体可以分解棉纤维表面的毛球正在开发中;一种对纤维损伤较小的护色的酶制剂进入实用阶段;一种可有效防止再污染,能与棉质衣料作用产生其他不同效果的独特酶制剂正在引起同行注意。今后,随着洗涤剂种类和用途的变化,用多种不同的酶制剂复配的复合型加酶洗涤剂品种将源源不断地上市,洗涤剂中酶制剂的用量不断增加、化学物质总量尽量降低将成为加酶洗涤剂发展的趋势。     

Highly effective adsorption performance of carboxymethyl cellulose microspheres crosslinked with epichlorohydrin

Different sizes of epichlorohydrin‐crosslinked carboxymethyl cellulose (E/CMC) microspheres were successfully prepared by an inverse suspension method. With further modification by monochloroacetic acid (MCA), MCA–E/CMC microspheres were successfully synthesized. The structures and morphologies of these microspheres were analyzed with polarizing microscopy, scanning electron microscopy, ultraviolet–visible spectroscopy, Fourier transform infrared spectroscopy, Brunauer–Emmett–Teller analysis, and laser particle ζ‐potential recording. The adsorption properties of the microspheres were investigated with methylene blue (MB) as a model pollutant. The highest adsorption amount of MB (998.2 mg/g) onto MCA–E_0.7/CMC which was the samples which treated with 0.7 mL of C₄H₉OH was obtained. Meanwhile, the effect of the operating parameters, such as the contact time, initial pH of the solution, temperature, and initial dye concentration, on the adsorption amount and MB removal were systematically studied. The results show that pseudo‐second‐order kinetic model provided the best correlation with the experimental data for the adsorption of MB onto the MCA–E_0.7/CMC microspheres. Both the physical and chemical adsorption played the main role in the adsorption process. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017 , 134, 44363.     

Preparation of Sodium carboxymethylcellulose/poly(methyl acrylate) IPN hydrogels and their application for adsorption

Sodium carboxymethylcellulose/poly(methyl acrylate) (NaCMC/PMA) interpenetrating polymer networks (IPNs) were prepared by fractional step in 40 wt % ethanol solution with N,N′‐methylenebisacrylamide as a crosslinker. Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and differential scanning calorimeter were used to characterize the NaCMC/PMA IPN hydrogels and confirm the IPN structure as well. Simultaneously, adsorption of the obtained IPN hydrogels to methylene blue (MB) was also investigated. It was observed that the adsorption of MB onto the hydrogels was mainly dependent on the initial concentration of MB and the pH of the solution. Adsorption rate of MB was much higher in the first 9 h than that in the following period and saturated adsorption amount of MB was 2370 mg/g at the initial MB concentration of 100 mg/L. Moreover, the adsorption capacity of the IPN hydrogels at the neutral pH condition is much higher than those at acid or alkaline pH conditions. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 41101.     

洗涤剂用酶的复合效应研究

以国产碱性蛋白酶、脂肪酶和纤维素酶为对象，研究了3种酶共存时蛋白酶对脂肪酶和纤维素酶活力的影响。结果表明，在实际洗涤液浓度下蛋白酶对脂肪酶和纤维素酶的活力并无明显负面影响。研究了3种酶在各种复合状态下的去污性能及其协同作用，结果显示上述3种酶的任何2种酶复合时均具有一定的协同效应，尤其是蛋白酶与脂肪酶或纤维素酶复合能获得显的协同效应。蛋白酶与脂肪酶或纤维素酶复配能使去污力相对提高50％以上。采用响应面分析法对蛋白酶／脂肪酶／纤维素酶三元复合酶的协同效应进行了优化，获得了其回归方程和最优组成。     

Dry action of Trichoderma reesei cellulases on cotton fabrics

Following treatment with Trichoderma reesei cellulases, the mechanical properties of cotton fabrics were measured to assess the effect of adsorbed enzymes. The ability of adsorbed cellulases to act as anchors for further wet finishing processes was studied. Dried fabric samples, after 105 days under usual storage conditions, showed neither significant strength loss, nor any changes in the degree of polymerisation. After five months of storage, the enzymes were still found to be active. The adsorption of T reesei cellulases produced an increase in staining levels after dyeing with an acid dye, but washing fastness was poor. Washing with soap under alkaline conditions removed proteins almost completely. Adsorption of T reesei cellulases is reversible and desorption increases from pH 5 to pH 10. Because of the protein desorption under usual alkaline washing conditions, no practical application of adsorbed cellulases for further finishing processes is suggested and more work is required.     

Production and characterization of a novel hierarchical porous silica adsorbent for removal of methylene blue dye from wastewaters

Adsorption of methylene blue (MB) dye by hierarchical porous silica (HPS) is reported as a means of removing the dye from wastewater. HPS was produced through a sol–gel process by templating on rubber particles in skimmed natural rubber latex. The silica precursor was tetraethyl orthosilicate and the costructure directing agent was 3-aminopropyltriethoxysilane. Based on nitrogen-sorption, the HPS had a total pore volume of 1.0?cm³/g, a specific surface area of 237?m²/g, and a peak pore size of 4?nm. Larger pores (sizes >100?nm to 1?µm) were observed by scanning electron microscopy. The pores had an ink-bottle morphology of narrow necks connected to large cavities. The adsorption obeyed the Langmuir isotherm and was endothermic. Pseudo-second-order adsorption kinetics were observed. The specific adsorption of MB on HPS was 0.225?mmol/g at equilibrium. HPS was superior to many adsorbents reported for removing MB from aqueous media.     

Characterization and photodegradation characteristics of organic dye for Pt–titania combined multi-walled carbon nanotube composite catalysts

Multi-walled carbon nanotubes (MWCNTs), titanium(IV) isopropoxide (TIP) and potassium hexachloroplatinate(IV) (K₂PtCl₆) were used for the preparation of Pt/MWCNT/TiO₂ composites. The composites were comprehensively characterized by Brauer–Emett–Teller surface area, scanning electron microscopy, transmission electron microscopy, X-ray diffraction, energy dispersive X-ray and UV–vis absorption spectroscopy. The photoactivity of the prepared materials under UV irradiation was tested using the conversion of methylene blue (MB) in aqueous solution. According to the results of MB removal experiment, it can be considered that the MB removal effect of the Pt/MWCNT/TiO₂ composites is affected by two kinds of effects: adsorption effect by MWCNTs and photocatalytic effect by TiO₂. Finally, the photocatalytic effect increases due to photo-induced-electron absorption effect by MWCNTs and electron trap effect by Pt metal.     

Enzyme immobilization on polyglycidylmethacrylate graft-copolymer of different polysaccharides

Comparative results obtained in preparing and characterizing samples of enzymes immobilized by reaction with polyglycidylmethacrylate (PGMA) copolymers with different polysaccharide matrices are reported. Sepharose copolymers having between 25 and 50% synthetic polymer were used to find the best immobilization conditions of horseradish peroxidase (HRP) and glucose-oxidase (GOD) (pH, time, temperature, enzyme cncentration). Activity, enzyme loading and coupling efficiency of immobilized HRP and GOD are greatly dependent on the type of matrix while the polymer content is less important. Coupling efficiencies between 0.8 and 1.5% have been obtained for HRP samples, whereas for GOD samples coupling efficiencies three times greater were obtained. HRP and GOD immobilized samples show K_m′ values greater than those of corresponding free enzymes and this indicates diffusion limitation phenomena. Storage, thermal and operational stability were also studied. In general the storage stability could be considered satisfactory (50% residual activity after 360 days). Sepharose and starch HRP-copolymers had an improved thermal stability compared with that of free enzyme. Residual activity found in continuous operation tests carried out on HRP-immobilized samples turned out to be dependent on support. HRP-PGMA-Cellulose sample gave the best results (50% residual activity after 16 days). PGMA-graft-copolymers have also been used to immobilize other enzymes such as α-amylase, α-chymotrypsin and cellulase.