鼠乳杆菌cnrz313苹果酸乳酸酶的纯化及性质研究。

Journal of applied biochemistry Pub Date : 1984-10-01
A M Strasser de Saad, A A Pesce de Ruiz Holgado, G Oliver
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引用次数: 0

摘要

对鼠乳杆菌的苹果酸乳酸酶进行了79倍纯化。经凝胶过滤和梯度凝胶电泳测定Mr = 22万。酶由两个明显相同的亚基(Mr = 110,000)组成,经十二烷基硫酸钠处理后观察到。NAD保护酶免于失活,在解离后加入NAD可以恢复酶的苹果酸乳酸活性。苹果酸盐、NAD和Mn2+的表观Km值分别为2.31 X 10(-2)、4.5 X 10(-4)和1.4 X 10(-4) mM。在37℃、pH为5.5、0.2 M磷酸盐缓冲液中酶活性最大。当pH值与最佳pH值相差很大时,观察到底物分子之间的正协同作用。温度大于30℃和小于30℃时,反应活化能分别为8000和16200 cal mol-1。苹果酸乳酸酶催化NAD和锰依赖反应l -苹果酸---- l -乳酸+ CO2。因此,该酶可以区别于众所周知的苹果酸酶[l-苹果酸:NAD+氧化还原酶,草酰乙酸脱羧(EC 1.1.1.38)或脱羧(EC 1.1.1.39)]。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Purification and properties of malolactic enzyme from Lactobacillus murinus CNRZ 313.

The malolactic enzyme of Lactobacillus murinus was purified 79 fold. Mr = 220,000 as determined by gel filtration and gradient gel electrophoresis. The enzyme consists of two apparently identical subunits (Mr = 110,000) that were observed after treatment with sodium dodecyl sulfate. NAD protected the enzyme against inactivation and its addition, after dissociation, restored the malolactic activity. The apparent Km's for malate, NAD, and Mn2+ were 2.31 X 10(-2), 4.5 X 10(-4), and 1.4 X 10(-4) mM, respectively. Maximum enzymatic activity was observed at 37 degrees C and pH 5.5 in 0.2 M phosphate buffer. At pH values substantially different from the optimum, a positive cooperativity between substrate molecules was observed. The activation energy of the reaction was 8000 and 16,200 cal mol-1 for the temperature values more than and less than 30 degrees C, respectively. Malolactic enzyme catalyzes the NAD and manganese-dependent reaction L-malate----L-lactate + CO2. Therefore, this enzyme can be distinguished from the well-known malic enzymes [L-malate: NAD+ oxidoreductase, oxaloacetate decarboxylating (EC 1.1.1.38) or decarboxylating (EC 1.1.1.39)].

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