Three different proteins exhibiting NAD-dependent acetaldehyde dehydrogenase activity from Alcaligenes eutrophus.
Three different proteins exhibiting NAD-dependent acetaldehyde dehydrogenase activity from Alcaligenes eutrophus.
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来自真养产碱菌的三种不同蛋白质表现出 NAD 依赖性乙醛脱氢酶活性。
DOI：
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发表时间：
1987
期刊：
影响因子：
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通讯作者：
H. Schlegel
中科院分区：
文献类型：
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作者：
D. Jendrossek;A. Steinbüchel;H. Schlegel
在有色链球藻中证实存在三种不同的蛋白，它们具有依赖于NAD的乙醛脱氢酶活性。发酵性酒精脱氢酶也具有乙醛脱氢酶活性，就是其中之一。用DEAE-Sephacel柱层析和三嗪染料亲和层析，从乙醇培养的真营养曲霉N9A突变株AS4中分离纯化了另外两种蛋白。乙醛脱氢酶II约占乙醇培养细胞中总可溶性蛋白质的14%，经纯化后均一。天然酶和亚基的相对分子质量分别为195,000和56,000。该酶对乙醛有很高的亲和力(Km=4微米)。乙醛脱氢酶I只占总可溶性蛋白质的不到1%。天然酶和亚基的相对分子质量分别为185,000和52,000。该酶对乙醛亲和力较低(Km=2.6 mM)。乙醛脱氢酶II的抗体与乙醛脱氢酶I不发生反应。两株不同的菌株，一株代表不同的突变类型，既能利用乙醇又能利用2，3-丁二醇，代表不同的突变体AS1；就像在AS4中一样，每个菌株的这些酶中的一种相当于细胞中总可溶性蛋白质的很大一部分。这些主要的乙醛脱氢酶都是从这两个菌株中纯化出来的；它们在所有相关性质上都与从AS4中分离的乙醛脱氢酶II相似。从AS4分离的酶的抗体与从AS1和TF93分离的酶有相同的交叉反应。
The existence of three different proteins exhibiting NAD-dependent acetaldehyde dehydrogenase activity was confirmed in Alicaligenes eutrophus. The fermentative alcohol dehydrogenase, which also exhibits acetaldehyde dehydrogenase activity, is one of these proteins. The other two proteins were purified from A. eutrophus N9A mutant AS4 grown on ethanol applying chromatography on DEAE-Sephacel and triazine-dye affinity media. Acetaldehyde dehydrogenase II, which amounts to about 14% of the total soluble protein in cells grown on ethanol, was purified to homogeneity. The relative molecular masses of the native enzyme and of the subunits were 195,000 or 56,000, respectively. This enzyme exhibits a high affinity for acetaldehyde (Km = 4 microM). Acetaldehyde dehydrogenase I amounts only to less than 1% of the total soluble protein. The relative molecular masses of the native enzyme and of the subunits were 185,000 and 52,000, respectively. This enzyme exhibits a low affinity for acetaldehyde (Km = 2.6 mM). Antibodies raised against acetaldehyde dehydrogenase II did not react with acetaldehyde dehydrogenase I. Two different strains, A. eutrophus N9A mutant AS1, which represents a different mutant type and can utilize both ethanol or 2,3-butanediol, and the type strain of A. eutrophus (TF93), which can utilize ethanol, form two acetaldehyde dehydrogenases during growth on ethanol, too. As in AS4, one of these enzymes from each strain amounted to a substantial portion of the total soluble protein in the cells. These major acetaldehyde dehydrogenases were purified from both strains; they resemble acetaldehyde dehydrogenase II isolated from AS4 in all relevant properties. Antibodies against the enzyme isolated from AS4 gave identical cross-reactions with the enzymes isolated from AS1 and TF93.