Cloning and expression of a pectate lyase from the oral spirochete Treponema pectinovorum ATCC 33768.
Cloning and expression of a pectate lyase from the oral spirochete Treponema pectinovorum ATCC 33768.
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口腔螺旋体果胶密螺旋体 ATCC 33768 果胶裂解酶的克隆和表达。
DOI：
10.1016/s0378-1097(03)00639-6
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发表时间：
2003
影响因子：
2.1
通讯作者：
Ryan,MariaE
中科院分区：
文献类型：
--
作者：
Walker,StephenG;Ryan,MariaE
从嗜果胶密螺旋体ATCC 33768中分离到一种果胶酸裂解酶基因，该基因是通过在大肠杆菌中异源表达粘粒文库获得的。体外转座子诱变鉴定了一个1293 bp的开放阅读框架，能够编码430个氨基酸的蛋白质，其预测的氨基末端信号序列为21个氨基酸。氨基酸序列的分析表明，它是多糖裂解酶家族10的成员，其中所有表征的成员均显示果胶酸裂解酶活性。弗罗姆中表达并纯化了氨基末端带His标签的重组形式PelA。杆菌该重组酶具有与其它细菌果胶酸裂解酶共同的特征，例如碱性pH最适、活性依赖于钙离子以及锌离子的抑制。
ThepelAgene, encoding a pectate lyase, fromTreponema pectinovorumATCC 33768 was isolated by heterologous expression of a cosmid library inEscherichia coli. In vitro transposon mutagenesis identified an open reading frame of 1293 bp capable of encoding a protein of 430 amino acids with a predicted amino-terminal signal sequence of 21 amino acids. Analysis of the amino acid sequence suggested that it is a member of the polysaccharide lyase family 10 of which all characterized members show pectate lyase activity. An amino-terminal His-tagged recombinant form of PelA was expressed and purified fromE. coli. The recombinant enzyme has characteristics common to other bacterial pectate lyases such as an alkaline pH optimum, dependence on calcium ions for activity, and inhibition by zinc ions.