[目的]以甲醛降解菌宛氏拟青霉菌F1-23为实验菌株,分离纯化得到甲醛脱氢酶的酶液,并考察其酶学性质。[方法]从液体培养液中收集菌体,通过细胞粉碎,采用(NH4)2SO4沉淀、透析、DFAE-Sepharose离子交换层析及Superdex-200凝胶过滤层析,纯...[目的]以甲醛降解菌宛氏拟青霉菌F1-23为实验菌株,分离纯化得到甲醛脱氢酶的酶液,并考察其酶学性质。[方法]从液体培养液中收集菌体,通过细胞粉碎,采用(NH4)2SO4沉淀、透析、DFAE-Sepharose离子交换层析及Superdex-200凝胶过滤层析,纯化得到电泳纯的FADH。[结果]该酶的分子量约为112.81 k Da;亚基分子量分别为65k Da和42 k Da;纯酶液活力为336.10 U/mg,较纯化前提高了4.02倍。对该酶的酶学性质分析结果表明,该酶最适反应温度为37℃,在25~45℃时稳定性很好;最适反应p H为8.0,在p H 6.5~8.0范围内酶活力较为稳定。底物特异性研究表明FADH最适底物为甲醛,相对偏好短链醛类。金属离子对酶活性的影响试验表明,Zn^(2+)、Mn^(2+)、Ag^+、Fe^(3+)和Hg^(2+)几乎完全抑制FADH酶活力;Ca^(2+)对酶活有促进作用。纯酶液在4℃环境下,在24h内可保存78%的活性。[结论]FADH结构较为特殊,且活性较大,为后期表达和深入研究其生物学功能提供理论和数据支持。展开更多
In this study, a carbon electrode electroless polymerization of polypyrrole, which formed a layer of conductive film interface,then absorped a layer of carbon nanotube particles by the way of self-assembled, is studie...In this study, a carbon electrode electroless polymerization of polypyrrole, which formed a layer of conductive film interface,then absorped a layer of carbon nanotube particles by the way of self-assembled, is studied. The modified electrode is used to electrostatic adherence of formaldehyde dehydrogenase, with Nafion solution capped. Moreover this paper discusses the work of electrode position,? PH value and scanning speed on the influence of the electrode response, the response potential is -1.2v, the optimal PH value is 7.5, and we also review the linear range of this electrode, we discovered taht this biosensor have a good linear relationship with the concentration of 1ug/ml-360ug/ml, the Correlation coefficient is 0.9983. In addition,with the stability of this electrode tested, it can be stored 15d at4°C.展开更多
文摘[目的]以甲醛降解菌宛氏拟青霉菌F1-23为实验菌株,分离纯化得到甲醛脱氢酶的酶液,并考察其酶学性质。[方法]从液体培养液中收集菌体,通过细胞粉碎,采用(NH4)2SO4沉淀、透析、DFAE-Sepharose离子交换层析及Superdex-200凝胶过滤层析,纯化得到电泳纯的FADH。[结果]该酶的分子量约为112.81 k Da;亚基分子量分别为65k Da和42 k Da;纯酶液活力为336.10 U/mg,较纯化前提高了4.02倍。对该酶的酶学性质分析结果表明,该酶最适反应温度为37℃,在25~45℃时稳定性很好;最适反应p H为8.0,在p H 6.5~8.0范围内酶活力较为稳定。底物特异性研究表明FADH最适底物为甲醛,相对偏好短链醛类。金属离子对酶活性的影响试验表明,Zn^(2+)、Mn^(2+)、Ag^+、Fe^(3+)和Hg^(2+)几乎完全抑制FADH酶活力;Ca^(2+)对酶活有促进作用。纯酶液在4℃环境下,在24h内可保存78%的活性。[结论]FADH结构较为特殊,且活性较大,为后期表达和深入研究其生物学功能提供理论和数据支持。
文摘In this study, a carbon electrode electroless polymerization of polypyrrole, which formed a layer of conductive film interface,then absorped a layer of carbon nanotube particles by the way of self-assembled, is studied. The modified electrode is used to electrostatic adherence of formaldehyde dehydrogenase, with Nafion solution capped. Moreover this paper discusses the work of electrode position,? PH value and scanning speed on the influence of the electrode response, the response potential is -1.2v, the optimal PH value is 7.5, and we also review the linear range of this electrode, we discovered taht this biosensor have a good linear relationship with the concentration of 1ug/ml-360ug/ml, the Correlation coefficient is 0.9983. In addition,with the stability of this electrode tested, it can be stored 15d at4°C.
