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首页> 外文期刊>Journal of bacteriology >Characterization of Activity and Expression of Isocitrate Lyase in Mycobacterium avium andMycobacterium tuberculosis
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Characterization of Activity and Expression of Isocitrate Lyase in Mycobacterium avium andMycobacterium tuberculosis

机译：鸟分枝杆菌和结核分枝杆菌中异柠檬酸裂合酶的活性和表达特征

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Analysis by two-dimensional gel electrophoresis revealed thatMycobacterium avium expresses several proteins unique to an intracellular infection. One abundant protein with an apparent molecular mass of 50 kDa was isolated, and the N-terminal sequence was determined. It matches a sequence in the M. tuberculosisdatabase (Sanger) with similarity to the enzyme isocitrate lyase of both Corynebacterium glutamicum and Rhodococcus fascians. Only marginal similarity was observed between this open reading frame (ORF) (termed icl) and a second distinct ORF (named aceA) which exhibits a low similarity to other isocitrate lyases. Both ORFs can be found as distinct genes in the various mycobacterial databases recently published. Isocitrate lyase is a key enzyme in the glyoxylate cycle and is essential as an anapleurotic enzyme for growth on acetate and certain fatty acids as carbon source. In this study we express and purify Icl, as well as AceA proteins, and show that both exhibit isocitrate lyase activity. Various known inhibitors for isocitrate lyase were effective. Furthermore, we present evidence that in both M. avium and M. tuberculosis the production and activity of the isocitrate lyase is enhanced under minimal growth conditions when supplemented with acetate or palmitate.
机译：二维凝胶电泳分析表明，鸟分枝杆菌表达了几种细胞内感染所特有的蛋白质。分离了一种表观分子量为50 kDa的丰富蛋白质，并确定了N端序列。它与 M中的序列匹配。结核病数据库（Sanger）与谷氨酸棒杆菌和法氏红球菌的异柠檬酸裂合酶相似。在该开放阅读框（ORF）（称为 icl ）和第二个不同的ORF（命名为 aceA ）之间仅观察到了边缘相似性，而后者与其他异柠檬酸裂合酶的相似性较低。在最近出版的各种分枝杆菌数据库中，两种ORF均可作为不同的基因被发现。异柠檬酸裂合酶是乙醛酸循环中的关键酶，对于在乙酸盐和某些脂肪酸作为碳源上的生长而言，它是一种不通尿的酶，必不可少。在这项研究中，我们表达并纯化Icl以及AceA蛋白，并显示两者均表现出异柠檬酸裂合酶活性。各种已知的异柠檬酸裂合酶抑制剂是有效的。此外，我们提供的证据表明，这两个 M。 avium 和 M。补充醋酸盐或棕榈酸盐，在最小的生长条件下，异柠檬酸裂合酶的生产和活性得到增强。

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