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融合酶
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  fusion enzyme
     This article focus on enzyme gene clone and heterogenesis expression,site directed modify and orthogenesis,fu-sion protein and fusion enzyme,enzyme mimics(abzyme and molecular imprinting)and telomere. It summarizes the ad-vances,trends and application of molecular enzyme engineering.
     本文着重介绍了酶基因克隆与异源表达、酶分子的定向改造和进化、融合蛋白与融合酶、酶的人工模拟(抗体酶、分子印迹技术)和端粒酶,综述了分子酶工程的研究进展、趋势及其应用。
短句来源
     Properties of the fusion enzyme of GST-CelA including the optimum temperature at 35℃ and the optimum pH about 7.2,showed that this fusion enzyme still belonged to cold-adapted enzyme and neutral enzyme.
     coli菌体破碎后 ,获取上清液 ,其中融合蛋白GST CelA浓度约为 78 5mg L。 分析了融合酶GST CelA的性质 ,其最适反应温度为 35℃ ,最适反应pH值为 7 2 ,为中性适冷酶。
短句来源
  “融合酶”译为未确定词的双语例句
     The construction of Thermotoga maritima endoglucanase Cel12B fused with CBD and the characterization of chimeric enzyme
     海栖热袍菌内切葡聚糖酶Cel12B与木聚糖酶XynA CBD结构域融合基因的构建、表达及融合酶性质分析
短句来源
     TB (with the N-terminal replacement mutation) further increase the half-life at 70 ℃ to 20 min. We optimized and combined the mutations mentioned above to obtain a new mutant TS (with replacement of N-terminal and introduction of a disulfide bridge).
     氨基端替换获得的融合酶TB,热稳定性较XYNB有很大的提高,在70℃的半衰期由XYNB的3min提高到20min。
短句来源
     Recent advances and applications of the molecular enzyme engineering are reviewed and discussed in this article.
     融合蛋白技术的发展使构建新型多功能融合酶成为可能。 这里对分子酶工程学的研究与发展情况进行了综述。
短句来源
  相似匹配句对
     Enzyme mimics
     模型
短句来源
     Medical image fusion
     医学图像融合
短句来源
     From Fission to Fusion
     从分裂到融合
短句来源
     The fusion protein exhibited good immunity andenzymatic activity.
     融合蛋白具有良好的免疫活性和一定的活性.
短句来源
     Cleavage of Two Fusion Proteins on Affinity Column
     两种融合蛋白的亲和柱上
短句来源
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  fusion enzyme
With respect to temperature stability, optimal pH and optimal temperature, substrate and stereospecificity, the purified fusion enzyme exhibited properties similar to those of the wild-type enzyme.
      
Starch-binding assays showed that the Kd and Bmax values for the fusion enzyme were 2.3?μM and 0.35?μmol/g, respectively.
      
The fusion enzyme, P450 2E1-BMR, contains the N-terminally modified residues 22-493 of the human P450 2E1 fused at the C-terminus to residues 473-1049 of the P450 BM3 reductase (BMR).
      
When packed in a column bioreactor, the immobilized fusion enzyme was able to completely degrade coumaphos up to a concentration of 0.2?mM.
      
A novel thermophilic fusion enzyme for trehalose production
      
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The developments of recombinant DNA technology and structural biology make it possible to modify enzyme in mole- cular level. Scientists show growing interests in the evolution or functional fusion of enzymes. Recent advances and applications of the molecular enzyme engineering are reviewed and discussed in this article.

酶工程的研究已经发展到分子水平 ,通过基因操作 ,已实现了许多酶的克隆和表达。定点突变成为研究酶结构与功能的常规手段 ,并被广泛用于改善酶的性能。体外分子进化方法则大幅提高了酶分子的进化效率 ,并有可能发展新功能酶。融合蛋白技术的发展使构建新型多功能融合酶成为可能。这里对分子酶工程学的研究与发展情况进行了综述。

To clone and overexpress the 3α hydroxysteroid dehydrogenase (3α HSD) in E.coli and to study its characterization, the 3α hydroxysteroid dehydrogenase gene was amplified by PCR from the genomic DNA of Comamonas testosteroni which was isolated from pond mud. The 3α HSD prokaryotic expression system was constructed with plasmid pET15b as the vector. The fusion protein with enzyme activity was overexpressed in the host bacteria of E.coli BL21(DE3) pLysS after IPTG induction. The fusion protein with...

To clone and overexpress the 3α hydroxysteroid dehydrogenase (3α HSD) in E.coli and to study its characterization, the 3α hydroxysteroid dehydrogenase gene was amplified by PCR from the genomic DNA of Comamonas testosteroni which was isolated from pond mud. The 3α HSD prokaryotic expression system was constructed with plasmid pET15b as the vector. The fusion protein with enzyme activity was overexpressed in the host bacteria of E.coli BL21(DE3) pLysS after IPTG induction. The fusion protein with an N terminal His tag sequence was purified in one step using metal chelate affinity chromatography to homogeneity on a Ni 2+ Sepharose column. The overall yield of the fusion protein was 64% and the relative activity was 194 7U/mg. After thrombin cleavage of the His tag, the molecular weight of the recombinant protein was 26 5kD according to the mass spectrum, which was identical to that predicted from the genomic sequence. With androsterone as the substrate and NAD + and thio NAD + as the cofactor, the optimum pH for the enzymatic reaction at 25℃ was 10 5 and 9 4 respectively. The rate of the enzymatic reaction was more quickly at 37℃ than at 25℃ and Q 10(25℃~35℃) was 1 7 The fusion 3α HSD could efficiently catalyze 3α hydroxyl dehydrogenation of androsterone and bile acids series with K m values in the range of 4 2~51 1μmol/L. The enzyme activity didn't need the participation of metal ions, but was inhibited by Fe 3+ , Fe 2+ and Zn 2+ . And EDTA did not affect its activity. The obtainment of fusion 3α HSD with high purity and activity and the investigation on its characterization laid a necessary foundation for the construction of a enzymatic cycling method to determine the human serum total bile acids.

从土壤中分离睾酮丛毛单胞菌 ,提取其基因组DNA ,PCR扩增 3α 羟类固醇脱氢酶 (3α HSD)基因。以质粒pET 1 5b为载体 ,构建 3α HSD的原核表达系统。经IPTG诱导后 ,得到了具有酶活性的融合蛋白 ,细菌总蛋白的表达量为 0 73g L ,其中融合酶蛋白占 1 6 4 % ,活性达 1 38× 1 0 5U L。利用融合蛋白N末端的His tag,经金属螯合亲和层析纯化后 ,得到了纯度较高的融合酶蛋白 ,酶蛋白的得率为 6 8% ,比活性为 1 94 7U mg。凝血酶切除His tag后 ,经质谱鉴定 ,重组蛋白的分子量为 2 6 5kD ,与理论推测值基本相同。 2 5℃以雄酮为底物 ,以NAD+ 和thio NAD+ 为辅因子时 ,融合蛋白参与酶促反应的最适pH分别为 1 0 5和 9 4。 37℃条件下酶促反应较快 ,2 5℃时的反应速度只有 37℃时的 5 2 % ,Q1 0 ( 2 5℃~ 35℃ ) 为 1 7。 2 5℃、pH1 0 5、以雄酮和各级胆汁酸为底物、NAD+ 为辅因子时 ,融合酶蛋白的动力学常数Km位于 ...

从土壤中分离睾酮丛毛单胞菌 ,提取其基因组DNA ,PCR扩增 3α 羟类固醇脱氢酶 (3α HSD)基因。以质粒pET 1 5b为载体 ,构建 3α HSD的原核表达系统。经IPTG诱导后 ,得到了具有酶活性的融合蛋白 ,细菌总蛋白的表达量为 0 73g L ,其中融合酶蛋白占 1 6 4 % ,活性达 1 38× 1 0 5U L。利用融合蛋白N末端的His tag,经金属螯合亲和层析纯化后 ,得到了纯度较高的融合酶蛋白 ,酶蛋白的得率为 6 8% ,比活性为 1 94 7U mg。凝血酶切除His tag后 ,经质谱鉴定 ,重组蛋白的分子量为 2 6 5kD ,与理论推测值基本相同。 2 5℃以雄酮为底物 ,以NAD+ 和thio NAD+ 为辅因子时 ,融合蛋白参与酶促反应的最适pH分别为 1 0 5和 9 4。 37℃条件下酶促反应较快 ,2 5℃时的反应速度只有 37℃时的 5 2 % ,Q1 0 ( 2 5℃~ 35℃ ) 为 1 7。 2 5℃、pH1 0 5、以雄酮和各级胆汁酸为底物、NAD+ 为辅因子时 ,融合酶蛋白的动力学常数Km位于 4 2~ 5 1 1 μmol L范围内。融合酶蛋白发挥催化作用并不需要金属离子的参与 ,而Fe3+ 、Fe2 + 、Zn2 + 则抑制酶活性 ,反应体系中加入EDTA也并不影响酶的活性。活性和纯度均较高的融合蛋白 3α HSD的获得及其生物学性质的研究 ,为血清总胆汁酸酶循环法测定的建立奠定了基础

With the development and application of gene engineering and protein project,enzyme engineering got highly advance.This article focus on enzyme gene clone and heterogenesis expression,site directed modify and orthogenesis,fu-sion protein and fusion enzyme,enzyme mimics(abzyme and molecular imprinting)and telomere.It summarizes the ad-vances,trends and application of molecular enzyme engineering.

随着基因工程和蛋白质工程的进展和应用,酶工程在分子水平上的研究与应用也得到了迅猛发展。本文着重介绍了酶基因克隆与异源表达、酶分子的定向改造和进化、融合蛋白与融合酶、酶的人工模拟(抗体酶、分子印迹技术)和端粒酶,综述了分子酶工程的研究进展、趋势及其应用。

 
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