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首页> 外文期刊>Journal of bacteriology >Biosynthetic Analysis of the Petrobactin Siderophore Pathway from Bacillus anthracis
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Biosynthetic Analysis of the Petrobactin Siderophore Pathway from Bacillus anthracis

机译：炭疽芽孢杆菌石油细菌铁载体途径的生物合成分析

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The asbABCDEF gene cluster from Bacillus anthracis is responsible for biosynthesis of petrobactin, a catecholate siderophore that functions in both iron acquisition and virulence in a murine model of anthrax. We initiated studies to determine the biosynthetic details of petrobactin assembly based on mutational analysis of the asb operon, identification of accumulated intermediates, and addition of exogenous siderophores to asb mutant strains. As a starting point, in-frame deletions of each of the genes in the asb locus (asbABCDEF) were constructed. The individual mutations resulted in complete abrogation of petrobactin biosynthesis when strains were grown on iron-depleted medium. However, in vitro analysis showed that each asb mutant grew to a very limited extent as vegetative cells in iron-depleted medium. In contrast, none of the B. anthracis asb mutant strains were able to outgrow from spores under the same culture conditions. Provision of exogenous petrobactin was able to rescue the growth defect in each asb mutant strain. Taken together, these data provide compelling evidence that AsbA performs the penultimate step in the biosynthesis of petrobactin, involving condensation of 3,4-dihydroxybenzoyl spermidine with citrate to form 3,4-dihydroxybenzoyl spermidinyl citrate. As a final step, the data reveal that AsbB catalyzes condensation of a second molecule of 3,4-dihydroxybenzoyl spermidine with 3,4-dihydroxybenzoyl spermidinyl citrate to form the mature siderophore. This work sets the stage for detailed biochemical studies with this unique acyl carrier protein-dependent, nonribosomal peptide synthetase-independent biosynthetic system.
机译：炭疽芽孢杆菌的 asbABCDEF 基因簇负责petrobactin的生物合成，petrobactin是一种儿茶酚型铁载体，在鼠的炭疽模型中起铁的吸收和毒力作用。我们基于 asb 操纵子的突变分析，鉴定积累的中间体以及向 asb 突变菌株添加外源铁载体，启动了研究以确定petrobactin组装的生物合成细节。首先，构建了 asb 基因座（ asbABCDEF ）中每个基因的框内缺失。当菌株在贫铁培养基上生长时，单个突变导致石蜡菌素生物合成的完全废除。但是，体外分析表明，每个 asb 突变体在缺铁培养基中作为营养细胞的生长都非常有限。相反， B都不存在。在相同的培养条件下，炭疽菌asb 突变株能够从孢子中生长出来。提供外源Petroproactin能够挽救每个 asb 突变株的生长缺陷。综上所述，这些数据提供了令人信服的证据，证明AsbA在生物合成Petrorobactin中进行了倒数第二步，涉及3,4-二羟基苯甲酰亚精胺与柠檬酸酯的缩合形成3,4-二羟基苯甲酰亚精胺柠檬酸酯。作为最后一步，数据显示AsbB催化3,4-二羟基苯甲酰基亚精胺柠檬酸酯的第二个分子与柠檬酸3,4-二羟基苯甲酰基亚精胺缩合形成成熟的铁载体。这项工作为这种独特的酰基载体蛋白依赖性，非核糖体肽合成酶依赖性生物合成系统奠定了进行详细生化研究的舞台。

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