[目的]海单胞菌Marinomonas sp.FW-1是1株经验证可以获得高活性芳基硫酸酯酶的菌株.为深入研究FW-1菌株产芳基硫酸酯酶机制,进一步筛选高活性的芳基硫酸酯酶基因片段,有必要解析FW-1菌株的全基因组序列信息.[方法]本研究采用高通量测序技术对FW-1进行全基因组测序,使用相关软件对测序数据进行基因组装、基因预测与功能注释、COG聚类分析等.结合异源表达的方法对其不同基因片段所产生的芳基硫酸酯酶活性进行分析.[结果]全基因组测序结果表明该基因组大小为3964876 bp,GC含量为44.03％,编码3590个蛋白基因,含有78个tRNA和25个rRNA操纵子.从全基因组测序结果中找到22个可能具有芳基硫酸酯酶活性的基因,对其中4个进一步异源表达后发现FW-1中至少含有的3个具有芳基硫酸酯酶活性的基因,其均含有芳基硫酸酯酶的特异性氨基酸基团C-X-P-X-R基团.[结论]本研究首次报道了1株含有多个芳基硫酸酯酶基因序列的菌株FW-1的全基因组序列,分析了基因组的基本特征,为芳基硫酸酯酶的进一步应用提供了思路.%[Objective] Marinomonas sp.FW-1 secretes high-activity arylsulfatase.This study was carried out to clone all the possible arylsulfatase in Marinomonas sp.FW-1 and investigate the motif information essential for arylsulfatase.[Methods] We used high-throughput sequencing to obtain the entire genome sequence of Marinomonas sp.FW-1.We carried out gene assembly,gene prediction,functional annotation and Clusters of Orthologous Groups clustering analysis.Thereafter,we cloned arylsulfatase genes in Escherichia coli and analyzed the activity of the recombinant proteins.[Results] The complete genome is of 3964876 bp in length,with an average Guanine and Cytosine content of 44.03％,encoding 3590 protein genes,containing 78 tRNA and 25 rRNA operons.Genome annotation indicated the presence of four possible arylsufatase genes.Among the four candidate genes,three genes contained C-X-P-X-R motif and their recombinant products were found to exhibit arylsulfatase activities.[Conclusion] This study revealed multiple arylsulfatase gene sequences in the complete genome Marinomonas sp.FW-1 strain,which predicted various potential applications of this strain.Moreover,C-X-P-X-R motif was confirmed to be essential for arylsulfatase activity.
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