Cloning and sequencing analysis of alginate lyase genes from the marine bacterium Vibrio sp O2
Cloning and sequencing analysis of alginate lyase genes from the marine bacterium Vibrio sp O2
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DOI：
10.1007/s10126-005-6157-z
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发表时间：
2006-09-01
影响因子：
3
通讯作者：
Matsuda, Hideyuki
中科院分区：
文献类型：
--
作者：
Kawamoto, Hitoshi;Horibe, Akio;Matsuda, Hideyuki
我们从日本三本关港的海水中分离出一种新的海洋细菌，该细菌在平板测定中显示出藻酸盐解聚活性。其中一株藻酸盐解聚菌的16 S核糖体RNA基因序列分析表明，该菌属于弧菌属，命名为弧菌属O2。克隆了弧菌O2的褐藻胶裂解酶基因，并在大肠杆菌中表达。获得了两个产褐藻胶裂解酶的克隆pVOA-A4和pVOA-B5。pVOA-A4的藻酸裂解酶基因alyVOA由一个858-bp的开放阅读框（ORF）组成，编码285个氨基酸残基，而pVOA-B5的alyVOB由一个828-bp的ORF组成，编码275个氨基酸残基。AlyVOA和AlyVOB的氨基酸序列与发光杆菌ATCC 43367藻酸盐多聚ManA裂解酶AlxM的同源性分别为92.3%和32.6%。在所有这三个序列中观察到对应于序列YFKAGXYXQ和RXELR的藻酸盐裂解酶共有区。AlyVOA和AlyVOB在平板测定中均降解聚甘露糖醛酸，因此被确认为聚（β-D-甘露糖醛酸）裂解酶。
We isolated a new marine bacteria, which displayed alginate-depolymerizing activity in plate assays, from seawater in Mihonoseki Harbor, Japan. Analysis of the 16S ribosomal RNA gene sequence of one of the isolates proved that this alginate-depolymerizing bacterium belonged to the genus Vibrio and it was named Vibrio sp. O2. The alginate lyase genes of Vibrio sp. O2 were cloned and expressed in Escherichia coli. Two alginate lyase-producing clones, pVOA-A4 and pVOA-B5, were obtained. The alginate lyase gene alyVOA from pVOA-A4 was composed of an 858-bp open reading frame (ORF) encoding 285 amino acid residues, while alyVOB from pVOA-B5 was composed of an 828-bp ORF encoding 275 amino acid residues. The degree of identity between the deduced amino acid sequences of AlyVOA or AlyVOB and Photobacterium sp. ATCC43367 alginate poly(ManA)lyase AlxM was 92.3% or 32.6%, respectively. Alginate lyase consensus regions corresponding to the sequences YFKAGXYXQ and RXELR were observed in all three of these sequences. AlyVOA and AlyVOB both degraded polymannuronate in plate assays and were therefore confirmed to be poly(beta-D-mannuronate)lyases.