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海洋蓝细菌聚球藻属菌株WH7805需要脲酶(尿素酰胺水解酶,EC 3.5.1.5)来利用尿素作为氮源:该酶的分子遗传学和生化分析

The marine cyanobacterium Synechococcus sp. WH7805 requires urease (urea amidohydrolase, EC 3.5.1.5) to utilize urea as a nitrogen source: molecular-genetic and biochemical analysis of the enzyme.

作者信息

Collier Jackie L, Brahamsha Bianca, Palenik Brian

出版信息

Microbiology (Reading). 1999 Feb;145 ( Pt 2):447-459. doi: 10.1099/13500872-145-2-447.

DOI:10.1099/13500872-145-2-447
PMID:10075427
Abstract

Cyanobacteria assigned to the genus Synechococcus are an important component of oligotrophic marine ecosystems, where their growth may be constrained by low availability of fixed nitrogen. Urea appears to be a major nitrogen resource in the sea, but little molecular information exists about its utilization by marine organisms, including Synechococcus. Oligonucleotide primers were used to amplify a conserved fragment of the urease (urea amidohydrolase, EC 3.5.1.5) coding region from cyanobacteria. A 5.7 kbp region of the genome of the unicellular marine cyanobacterium Synechococcus sp. strain WH7805 was then cloned, and genes encoding three urease structural subunits and four urease accessory proteins were sequenced and identified by homology. The WH7805 urease had a predicted subunit composition typical of bacterial ureases, but the organization of the WH7805 urease genes was unique. Biochemical characteristics of the WH7805 urease enzyme were consistent with the predictions of the sequence data. Physiological data and sequence analysis both suggested that the urease operon may be nitrogen-regulated by the ntcA system in WH7805. Inactivation of the large subunit of urease, ureC, prevented WH7805 and Synechococcus WH8102 from growing on urea, demonstrating that the urease genes cloned are essential to the ability of these cyanobacteria to utilize urea as a nitrogen source.

摘要

属于聚球藻属的蓝细菌是贫营养海洋生态系统的重要组成部分,在这些生态系统中,它们的生长可能受到固定氮供应不足的限制。尿素似乎是海洋中的主要氮源,但关于包括聚球藻在内的海洋生物对其利用的分子信息却很少。使用寡核苷酸引物从蓝细菌中扩增脲酶(尿素酰胺水解酶,EC 3.5.1.5)编码区的一个保守片段。然后克隆了单细胞海洋蓝细菌聚球藻属菌株WH7805基因组的一个5.7 kbp区域,并对编码三个脲酶结构亚基和四个脲酶辅助蛋白的基因进行了测序,并通过同源性进行了鉴定。WH7805脲酶具有细菌脲酶典型的预测亚基组成,但WH7805脲酶基因的组织方式是独特的。WH7805脲酶的生化特性与序列数据的预测一致。生理数据和序列分析均表明,脲酶操纵子可能受WH7805中ntcA系统的氮调节。脲酶大亚基ureC的失活阻止了WH7805和聚球藻WH8102在尿素上生长,表明克隆的脲酶基因对于这些蓝细菌利用尿素作为氮源的能力至关重要。

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