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A New Homo-Hexamer Mn-Containing Catalase from Geobacillus sp. WCH70

机译:芽孢杆菌属的一种新的含六方体锰过氧化氢酶WCH70

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摘要

Catalase is an effective biocatalyst to degrade hydrogen peroxide to water and oxygen that can serve in textile effluent treatment to remove residual H 2 O 2 . Thermostable catalases are needed to withstand both the high temperature and pH of textile wastewater. We have cloned the Mn-containing catalase gene ACS24898.1 from Geobacillus sp. WCH70, which originated from thermophilic organisms, and expressed it in Escherichia coli in activated form. The recombinant protein has been purified to homogeneity and identified to be a new homo-hexamer Mn-containing catalase. The native molecular mass of the catalase has been measured to be 138 kDa by size-exclusion chromatography. The new enzyme has optimum catalyzed activity at pH 9.0 and a temperature of 75 °C. It is thermostable up to 70 °C for 8 h incubation and maintains 80% and 50% activity, respectively, at 80 °C after 5 h and 90 °C after 1 h. At 75 °C and pH 9.0, the K m is 67.26 mM for substrate H 2 O 2 and the rate of reaction at H 2 O 2 saturation, V max , is 75,300 U/mg. The thermophilic and alkaline preferred properties of this new Mn-catalase are valuable features in textile wastewater treatment.
机译:过氧化氢酶是一种有效的生物催化剂,可将过氧化氢降解为水和氧气,可用于纺织品废水处理中以去除残留的H 2 O 2。需要耐热的过氧化氢酶以承受纺织品废水的高温和pH值。我们已经从Geobacillus sp。克隆了含Mn的过氧化氢酶基因ACS24898.1。 WCH70,起源于嗜热生物,并以活化形式在大肠杆菌中表达。重组蛋白已纯化至同质,并被鉴定为一种新的含锰的均六聚体过氧化氢酶。通过尺寸排阻色谱法测得过氧化氢酶的天然分子量为138kDa。这种新酶在pH 9.0和75°C的温度下具有最佳的催化活性。它在高达70°C的温度下孵育8小时具有热稳定性,并在5 h后的80°C和1 h后的90°C下分别保持80%和50%的活性。在75°C和pH 9.0下,底物H 2 O 2的K m为67.26 mM,在H 2 O 2饱和时的反应速率V max为75,300 U / mg。这种新的Mn-过氧化氢酶的嗜热和碱性优选特性是纺织品废水处理中的宝贵功能。

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