SEQUENCE OF THE ALGL GENE OF PSEUDOMONAS-AERUGINOSA AND PURIFICATION OF ITS ALGINATE LYASE PRODUCT
SEQUENCE OF THE ALGL GENE OF PSEUDOMONAS-AERUGINOSA AND PURIFICATION OF ITS ALGINATE LYASE PRODUCT
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DOI：
10.1016/0378-1119(93)90662-m
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发表时间：
1993-09-06
期刊：
影响因子：
3.5
通讯作者：
CHAKRABARTY, AM
中科院分区：
文献类型：
--
作者：
BOYD, A;GHOSH, M;CHAKRABARTY, AM
铜绿假单胞菌的海藻酸盐裂解酶编码基因（algL）定位在染色体上34分钟海藻酸盐生物合成基因簇中1.7 kb的EcoRI-XbaI片段上。该DNA片段的核苷酸序列显示了一个编码M(r) 40 885蛋白的ORF，该蛋白与生物合成基因簇内其他藻类基因的转录方向相同。预测的蛋白有一个潜在的n端信号肽，这与其提出的质周位置一致。在大肠杆菌中过量产生AlgL蛋白并纯化。纯化后的蛋白具有海藻酸裂解酶活性。此外，还构建了粘液样P. aeruginosa 8830的algL插入突变体。该突变体（alm 1）具有非黏液样表型，这是由于一个重要的藻类基因algA转录的极性效应。因此，algL基因位于藻酸盐生物合成基因簇的一个区域内，该区域对藻酸盐的生产似乎不是必需的。
The alginate lyase-encoding gene (algL) of Pseudomonas aeruginosa was localized to a 1.7-kb EcoRI-XbaI fragment within the alginate biosynthetic gene cluster at 34 minutes on the chromosome. The nucleotide sequence of this DNA fragment revealed an ORF encoding a protein of M(r) 40 885 which is transcribed in the same orientation as the other alg genes within the biosynthetic gene cluster. The predicted protein has a potential N-terminal signal peptide which is consistent with its proposed periplasmic location. The AlgL protein was overproduced in Escherichia coli and purified. The purified protein was shown to have alginate lyase activity. In addition, an algL insertion mutant of the mucoid P. aeruginosa 8830 was constructed. This mutant (alm 1) had a nonmucoid phenotype due to a polar effect on the transcription of an essential alg gene, algA. Thus, the algL gene is located within a region of the alginate biosynthetic gene cluster that appears to be non-essential for alginate production.