共查询到19条相似文献,搜索用时 312 毫秒
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在反应温度为45℃、反应时间为6 h、丙烯酰胺(AM)质量浓度为40 g/L、引发剂用量为单体质量1%的条件下制得PAM-SiO2复合载体,利用红外光谱表征了其化学结构,热失重法测定其接枝量为21%。在所制PAM-SiO2复合载体上固载青霉素酰化酶,其固载最适条件为:戊二醛用量0.1 mmol/L,pH值7.64,给酶量1%,温度42℃,时间11 h~12 h。此条件下所得固定化酶的活力为34000 U/mL;测得所制固定化酶的最适pH值为7.82,最适温度为45℃。 相似文献
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分子筛固定氨基酰化酶Ⅰ的研究 总被引:2,自引:0,他引:2
以Y型分子筛为载体,分别采用吸附法、吸附-戊二醛交联法、偶联法和偶联-重氮法4种方法对氨基酰化酶I进行固定化。 研究发现偶联法获得的分子筛固定化酶的活性最高,达0.258 IUmg-1,并对分子筛固定化酶最适pH值、最适反应温度、重复使用性和米氏常数进行了测定;结果表明该载体适合氨基酰化酶I的固定,可以获得较高的酶活力,分子筛固定化酶的最适pH值、最适反应温度都有所拓宽。 相似文献
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以Y型分子筛为载体,分别采用吸附法、吸附-戊二醛交联法、偶联法和偶联-重氮法4种方法对氨基酰化酶I进行固定化。 研究发现偶联法获得的分子筛固定化酶的活性最高,达0.258 IUmg-1,并对分子筛固定化酶最适pH值、最适反应温度、重复使用性和米氏常数进行了测定;结果表明该载体适合氨基酰化酶I的固定,可以获得较高的酶活力,分子筛固定化酶的最适pH值、最适反应温度都有所拓宽。 相似文献
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以氨基树脂为载体对S-腺苷甲硫氨酸(SAM)合成酶进行固定化,优化了酶的固定化条件并对固定化酶的性质进行了研究。优化的固定化条件为:戊二醛体积分数5%、SAM合成酶添加量20mg·g-1、固定化时间5h。所制备的固定化SAM合成酶的酶活力为476.8U·g-1,酶活力回收率为74.5%。与游离SAM合成酶相比,固定化SAM合成酶的稳定性大幅提高,在50℃孵育5h酶活力仍保留61.2%,而游离SAM合成酶则完全失活;在pH值为6.0~6.5、8.0~9.5的缓冲溶液中,固定化SAM合成酶也更加稳定;固定化SAM合成酶连续催化反应10批次,酶活力保留86.3%;固定化SAM合成酶在4℃储存30d,酶活力保留81.4%。固定化SAM合成酶米氏常数KATPm=0.14mmol·L-1,KLm-Met=0.28mmol·L-1。 相似文献
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以壳聚糖为载体固定化青霉素酰化酶的研究 总被引:12,自引:1,他引:11
介绍了以壳聚糖为载体固定化青霉素酰化酶需首先制备壳聚糖颗粒 ,使用戊二醛、甲醛、乙二醛 3种活化剂处理所得的壳聚糖颗粒 ,确定了以戊二醛为活化剂交联其上的氨基共价结合青霉素酰化酶的固定化方法。从戊二醛的浓度、pH值、固定化时间、固定化pH值、酶用量等条件摸索了最佳固定化条件 ,获得了酶活力为4 0 0 0 0U/ (g·h)、回收率为 5 0 %左右的固定化青霉素酰化酶。 相似文献
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用分相法和填充法分别制作了两种载体多孔玻璃微珠FXBL和TCBL,用共价偶联法分别固定α-淀粉酶,确定了其最佳固定条件和最佳应用条件,并研究了固定化酶的性质。主要结论如下:①最佳固定条件为:温度10℃;pH=6.2;给酶量TCBL 2.6 g/LF、XBL 2.4 g/L;时间12 h;②最佳应用条件为:温度75℃,比自由酶高5℃;pH值TCBL固定化酶为5.2、FXBL固定化酶为5.4,分别比自由酶的最适pH值6.0低0.8和0.6个pH值单位;③固定化酶的主要性质为:在80℃受热1 h,固定化酶的活力下降小于7%,而自由酶活力则下降至63%;FXBL固定化酶和TCBL固定化酶分别使用5次和8次还可以保持60%的活力。 相似文献
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为提高青霉素酰化酶的催化性能和热稳定性,在酶组装过程中添加小分子试剂对介孔泡沫硅载体表面过量的活化位点进行封闭。详细考察了小分子添加质量分数和种类对青霉素酰化酶负载率、催化活力及热稳定性的影响。实验结果得到:经精氨酸封闭的固定化酶活力提高至1.92倍;甘氨酸封闭的固定化酶5 h的50℃热稳定性提高至2.9倍,甘氨酸和谷氨酸封闭的固定化酶50℃热处理25 h仍保持87.9%和82.2%的残余活力;甘氨酸和谷氨酸封闭的固定化酶最适催化pH值向中性偏移且对pH值的耐受性增强。结果表明,在青霉素酰化酶共价组装过程中添加合适的小分子封闭能显著提高酶的催化性能和热稳定性。 相似文献
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Feng Wang Chen Guo Hui‐Zhou Liu Chun‐Zhao Liu 《Journal of chemical technology and biotechnology (Oxford, Oxfordshire : 1986)》2008,83(1):97-104
BACKGROUND: Immobilized enzymes provide many advantages over free enzymes including repeated or continuous reuse, easy separation of the product from reaction media, easy recovery of the enzyme, and improvement in enzyme stability. In order to improve catalytic activity of laccase and increase its industrial application, there is great interest in developing novel technologies on laccase immobilization. RESULTS: Magnetic Cu2+‐chelated particles, prepared by cerium‐initiated graft polymerization of tentacle‐type polymer chains with iminodiacetic acid (IDA) as chelating ligand, were employed for Pycnoporus sanguineus laccase immobilization. The particles showed an obvious high adsorption capacity of laccase (94.1 mg g?1 support) with an activity recovery of 68.0% after immobilization. The laccase exhibited improved stability in reaction conditions over a broad temperature range between 45 °C and 70 °C and an optimal pH value of 3.0 after being adsorbed on the magnetic metal‐chelated particles. The value of the Michaelis constant (Km) of the immobilized laccase (1.597 mmol L?1) was higher than that of the free one (0.761 mmol L?1), whereas the maximum velocity (Vmax) was lower for the adsorbed laccase. Storage stability and temperature endurance of the immobilized laccase were found to increase greatly, and the immobilized laccase retained 87.8% of its initial activity after 10 successive batch reactions. CONCLUSION: The immobilized laccase not only can be operated magnetically, but also exhibits remarkably improved catalytic capacity and stability properties for various parameters, such as pH, temperature, reuse, and storage time, which can provide economic advantages for large‐scale biotechnological applications of laccase. Copyright © 2007 Society of Chemical Industry 相似文献
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用硬模板法制得具有三维连续贯通孔道结构的整体型大孔/介孔SiO2,通过多巴胺(DA)在大孔/介孔SiO2孔道表面的原位氧化聚合,制得聚多巴胺(PDA)功能化修饰的整体型大孔/介孔复合材料(PDA/SiO2)。应用SEM、BET、FTIR和TG等技术对修饰前后的材料进行表征。以PDA/SiO2为载体固定诺维信工业级漆酶,系统研究了pH、固定化时间、漆酶初始浓度及温度对漆酶固定化的影响;以偶氮荧光桃红作为模拟污染物,研究了固定化漆酶对染料的催化降解性能。结果显示,在漆酶浓度为80mg/mL、pH为4.0、固定化时间为6h及固定化温度为25℃时,固定化漆酶酶活达到最高(348.9U/g)。在偶氮荧光桃红浓度为10mg/L、pH为7.0、温度为30℃、降解时间为8h时,固定化漆酶对偶氮荧光桃红脱色率 ≥ 99.9%,且固定化漆酶易从反应体系中分离,重复使用性能良好。 相似文献
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以醋酸纤维素(CA)为原料,采用溶液浇铸法成膜。用物理包埋法将荧光指示剂固定CA膜中。以戊二醛为交联剂,将葡萄糖氧化酶固定于膜表面。研究了高碘酸钠、乙二胺、戊二醛、给酶量、pH值等固定化条件对酶活的影响,确定了优化的固定条件:在室温下与0.5 mol/L的高碘酸钠反应30 min;与0.04 mol/L的乙二胺反应150 min时;与溶度为1.5%(v/v)的戊二醛交联120 min;在4℃条件下,在pH6.5、给酶量35 mg/mL的酶液中交联21 h。利用SEM分析表明,荧光指示剂均匀分布膜中,成膜质量较好。通过优化固定条件,把荧光指示剂和葡萄糖氧化物酶(GOD)同时固定醋酸纤维素膜上,可以得到同时具有光敏感性和酶催化能力的复合敏感膜。 相似文献
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Jung Hee Park Huahua Xue Jin Suk Jung Keungarp Ryu 《Korean Journal of Chemical Engineering》2012,29(10):1409-1412
Laccase from Trametes versicolor was readily immobilized on carbon nanomaterials including multiwalled carbon nanotubes (MWNTs), carboxylated multiwalled carbon nanotubes (MWNT-COOHs), and graphene oxides (GOs), by physical adsorption without using coupling agents. The immobilized amount of laccase strongly depends on the pH of the aqueous buffers of the immobilization mixture. As the pH of the aqueous buffer for immobilization increases, the immobilized amount of laccase decreases. The activity of the immobilized laccase on the three carbon nanomaterials exhibits a bell-shaped dependence on the pH of the immobilization solution with maximum activity at pH 6 or 7. When the immobilization solution becomes acidic or basic, the activity of the immobilized laccase declines significantly. The amount and the activity of immobilized laccase were maximum for graphene oxides as substrate material for immobilization. 相似文献
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Qingqing Wang Lin Peng Yuanzhi Du Jing Xu Yibing Cai Quan Feng Fenglin Huang Qufu Wei 《Journal of Porous Materials》2013,20(3):457-464
The electrospun PMMA/O-MMT microfibrous membranes were pretreated by oxygen plasma to create substrates with better adsorption capability. The amount of O-MMT and conditions of plasma treatment were optimized for maximum laccase immobilization on these pretreated surfaces of the microfibrous membranes. The surface morphology and chemistry of the composite microfibrous membranes after plasma treatment and laccase immobilization were investigated by SEM and FTIR. The immobilized laccase showed better resistance to pH and temperature changes than that of the free form laccase, and after 10 successive runs of repeated use, the immobilized laccase still retained 30 % of its initial activity. Reactive X-3B was successfully degraded by both free and immobilized laccase. 相似文献
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以溶菌酶作为诱导剂,仿生合成了ZrO2固定化漆酶纳米颗粒,其酶活回收率达59%,采用场发射扫描电子显微镜(FESEM)、能谱仪(EDS)、热重分析仪(TGA)等手段对ZrO2纳米颗粒及ZrO2固定化漆酶颗粒进行表征,结果表明漆酶可成功固定到ZrO2颗粒中,同时还证明了溶菌酶既作为诱导剂催化ZrO2的形成,又作为生物模板同酶一起包埋在ZrO2颗粒中。固定化漆酶的最适pH为3,最适温度为70℃,相比于游离酶,其pH、温度稳定性都有明显提高;固定化漆酶纳米颗粒在4℃下储存30d,活性为初始酶活的95%,重复使用5次,固定化酶的残余酶活力仍有60%。此外,固定化漆酶在6h内对孔雀石绿染料的脱色率高达95%以上,通过紫外-可见吸收光谱分析(UV-vis)可知,固定化漆酶对孔雀石绿染料的处理是由吸附和降解联合作用引起的脱色。 相似文献
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以聚丙烯酸(PAA)改性的聚乙烯(PE)膜为载体,研究了醇脱氢酶(ADH)的两种固定化路线,并以甲醛为底物考察了固定化酶的催化性能。路线1用聚乙烯亚胺(PEI)进一步改性,使用戊二醛(GA)固定化ADH。最优固定化pH为6.0,温度为5~15℃,酶浓度为1.0 mg/ml,GA浓度为0.01%(质量);固定化酶的最适反应pH为6.5,温度为15~30℃,反应速率最高为9.6 μmol/(L·min);重复利用10次后可保持47.3%的活性。路线2以PAA-PE为载体,用1-(3-二甲氨基丙基)-2-乙基碳二亚胺盐酸盐(EDC)和N-羟基琥珀酰亚胺(NHS)为活化剂,固定化ADH。EDC和NHS最优摩尔比为1∶0.5,固定化时间为24 h;固定化酶的最适反应pH为6.5,温度为20~37℃,反应速率为15.58 μmol/(L·min);重复利用10次后可保持53.8%的活性。 相似文献
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Qingqing Wang Lin Peng Guohui Li Ping Zhang Dawei Li Fenglin Huang Qufu Wei 《International journal of molecular sciences》2013,14(6):12520-12532
The TiO2-montmorillonite (TiO2-MMT) complex was prepared by blending TiO2 sol and MMT with certain ratio, and its properties as an enzyme immobilization support were investigated. The pristine MMT and TiO2-MMT calcined at 800 °C (TiO2-MMT800) were used for comparison to better understand the immobilization mechanism. The structures of the pristine MMT, TiO2-MMT, and TiO2-MMT800 were examined by HR-TEM, XRD and BET. SEM was employed to study different morphologies before and after laccase immobilization. Activity and kinetic parameters of the immobilized laccase were also determined. It was found that the TiO2 nanoparticles were successfully introduced into the MMT layer structure, and this intercalation enlarged the “d value” of two adjacent MMT layers and increased the surface area, while the calcination process led to a complete collapse of the MMT layers. SEM results showed that the clays were well coated with adsorbed enzymes. The study of laccase activity revealed that the optimum pH and temperature were pH = 3 and 60 °C, respectively. In addition, the storage stability for the immobilized laccase was satisfactory. The kinetic properties indicated that laccase immobilized on TiO2-MMT complexes had a good affinity to the substrate. It has been proved that TiO2-MMT complex is a good candidate for enzyme immobilization. 相似文献
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Using polyacrylic acid (PAA) modified polyethylene (PE) membrane as a carrier, two immobilization routes of alcohol dehydrogenase (ADH) were studied, and the catalytic performance of immobilized enzyme was investigated using formaldehyde as a substrate. In the first route, PAA-PE membrane was further modified by polyethyleneimine (PEI) and then ADH was covalently linked by glutaraldehyde (GA) to the surface of PEI/PAA-PE. The results show that the optimal immobilization pH was 6.0, immobilization temperature was 5—15℃, ADH and GA concentrations were 1.0mg/ml and 0.01%(mass). For immobilized enzyme, the optimal reaction pH was 6.5, temperature was 15—30℃, and the highest reaction rate was 9.6 μmol/(L·min), the remaining activity was 47.3% after 10 use cycles. In the second route, ADH was immobilized on PAA-PE membrane with 1-(3-dimethylaminopropyl)-2-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) as activators. The results show that the optimal molar ratio of EDC and NHS was 1∶0.5, and the immobilization time was 24 h. For immobilized enzyme, the optimal reaction pH was 6.5, temperature was 20—37℃, and the highest reaction rate was 15.58 μmol/(L·min), 53.8% activity was remained after 10 cycles. 相似文献