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首页> 外文期刊>Infection and immunity >Complete nucleotide sequence of the structural gene for alkaline proteinase from Pseudomonas aeruginosa IFO 3455.
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Complete nucleotide sequence of the structural gene for alkaline proteinase from Pseudomonas aeruginosa IFO 3455.

机译：铜绿假单胞菌IFO 3455碱性蛋白酶结构基因的完整核苷酸序列。

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摘要

The DNA-encoding alkaline proteinase (AP) of Pseudomonas aeruginosa IFO 3455 was cloned, and its complete nucleotide sequence was determined. When the cloned gene was ligated to pUC18, the Escherichia coli expression vector, the gene-incorporated bacteria expressed high levels of both AP activity and AP antigens. The amino acid sequence deduced from the nucleotide sequence revealed that the mature AP consists of 467 amino acids with a relative molecular weight of 49,507. The amino acid composition predicted from the DNA sequence was similar to the chemically determined composition of purified AP reported previously. The amino acid sequence analysis revealed that both the N-terminal side sequence of the purified AP and several internal lysyl peptide fragments were identical to the deduced amino acid sequences. The percent homology of amino acid sequences between AP and Serratia protease was about 55%. The zinc ligands and an active site of the AP were predicted by comparing the structure of the enzyme with of Serratia protease, thermolysin, Bacillus subtilis neutral protease, and Pseudomonas elastase.
机译：克隆了铜绿假单胞菌IFO 3455的DNA编码碱性蛋白酶（AP），并确定了其完整核苷酸序列。当将克隆的基因与大肠杆菌表达载体pUC18连接时，掺有基因的细菌表达高水平的AP活性和AP抗原。从核苷酸序列推导的氨基酸序列显示，成熟的AP由467个氨基酸组成，相对分子量为49,507。由DNA序列预测的氨基酸组成与先前报道的纯化AP的化学测定的组成相似。氨基酸序列分析表明，纯化的AP的N末端侧序列和几个内部赖氨酰肽片段均与推导的氨基酸序列相同。 AP和沙雷氏菌蛋白酶之间的氨基酸序列同源性百分比为约55％。通过比较该酶与沙雷氏菌蛋白酶，嗜热菌蛋白酶，枯草芽孢杆菌中性蛋白酶和假单胞菌弹性蛋白酶的结构，可以预测AP的锌配体和活性位点。

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