建立了一种基于荷移反应的高效液相色谱测定氨基丁醇含量的分析方法。在p H 8.4的硼砂-硼酸缓冲溶液中,氨基丁醇与四氯苯醌于60℃反应60 min,利用高效液相色谱法-紫外检测器进行分析。荷移络合物采用Agilent Ex tend C18色谱柱(250 m m&...建立了一种基于荷移反应的高效液相色谱测定氨基丁醇含量的分析方法。在p H 8.4的硼砂-硼酸缓冲溶液中,氨基丁醇与四氯苯醌于60℃反应60 min,利用高效液相色谱法-紫外检测器进行分析。荷移络合物采用Agilent Ex tend C18色谱柱(250 m m×4.6 m m,5μm)分离,以0.001%(体积分数)三乙胺甲醇溶液为流动相进行梯度洗脱,流速为1 m L/min,检测波长为350 nm。该方法对氨基丁醇的定量限为0.01 g/L,线性范围为0.1~0.6g/L,相关系数(R^2)为0.999 4;方法的加标回收率为98.3%~103.6%,相对标准偏差(RSD)为0.9%~1.6%。该法简便快捷,适用于氨基丁醇含量的快速检测。展开更多
1,3-propanediol (1,3-PD) is an important material for chemical industry,and there has been always much interest in the production of 1,3-PD using all possible routes. The genes encoding glyc-erol dehydratase (GDHt) fr...1,3-propanediol (1,3-PD) is an important material for chemical industry,and there has been always much interest in the production of 1,3-PD using all possible routes. The genes encoding glyc-erol dehydratase (GDHt) from Citrobacter freundii,Klebsiella pneumoniae and metagenome were cloned and expressed in E. coli. All glycerol dehy-dratases but the one from metagenome could be detected to show enzyme activities. In order to im-prove the enzymatic properties of GDHts,the genes encoding α and β-γ subunits were cloned,and the enzyme characteristics were evolved by rational de-sign based on their 3D structures which were con-structed by homology modeling. Six heteroenzymes were obtained by swapping the α subunit genes of these three different-source-derived GDHts. The pH,thermal stability and Vmax of some heteroenzymes were dramatically improved by 2―5 times compared with the wild one (GDHtKP). The GDHt cloned from metagenome,originally proved to be with no enzyme activity,was converted into active enzyme by swap-ping its subunits with other different GDHts. In addi-tion,the effect of subtle 3D structural changes on the properties of the enzyme was also observed.展开更多
文摘建立了一种基于荷移反应的高效液相色谱测定氨基丁醇含量的分析方法。在p H 8.4的硼砂-硼酸缓冲溶液中,氨基丁醇与四氯苯醌于60℃反应60 min,利用高效液相色谱法-紫外检测器进行分析。荷移络合物采用Agilent Ex tend C18色谱柱(250 m m×4.6 m m,5μm)分离,以0.001%(体积分数)三乙胺甲醇溶液为流动相进行梯度洗脱,流速为1 m L/min,检测波长为350 nm。该方法对氨基丁醇的定量限为0.01 g/L,线性范围为0.1~0.6g/L,相关系数(R^2)为0.999 4;方法的加标回收率为98.3%~103.6%,相对标准偏差(RSD)为0.9%~1.6%。该法简便快捷,适用于氨基丁醇含量的快速检测。
文摘1,3-propanediol (1,3-PD) is an important material for chemical industry,and there has been always much interest in the production of 1,3-PD using all possible routes. The genes encoding glyc-erol dehydratase (GDHt) from Citrobacter freundii,Klebsiella pneumoniae and metagenome were cloned and expressed in E. coli. All glycerol dehy-dratases but the one from metagenome could be detected to show enzyme activities. In order to im-prove the enzymatic properties of GDHts,the genes encoding α and β-γ subunits were cloned,and the enzyme characteristics were evolved by rational de-sign based on their 3D structures which were con-structed by homology modeling. Six heteroenzymes were obtained by swapping the α subunit genes of these three different-source-derived GDHts. The pH,thermal stability and Vmax of some heteroenzymes were dramatically improved by 2―5 times compared with the wild one (GDHtKP). The GDHt cloned from metagenome,originally proved to be with no enzyme activity,was converted into active enzyme by swap-ping its subunits with other different GDHts. In addi-tion,the effect of subtle 3D structural changes on the properties of the enzyme was also observed.
