Lambert Emily A, Popham David L
Department of Biological Sciences, Virginia Tech, Life Sciences I-MC0910, Washington St., Blacksburg, VA 24061, USA.
J Bacteriol. 2008 Dec;190(23):7601-7. doi: 10.1128/JB.01054-08. Epub 2008 Oct 3.
Bacillus anthracis spores, the infectious agents of anthrax, are notoriously difficult to remove from contaminated areas because they are resistant to many eradication methods. These resistance properties are due to the spore's dehydration and dormancy and to the multiple protective layers surrounding the spore core, one of which is the cortex. In order for B. anthracis spores to germinate and resume growth, the cortex peptidoglycan must be depolymerized. This study reports on analyses of sleL (yaaH), which encodes a cortex-lytic enzyme. The inactivation of sleL does not affect vegetative growth, spore viability, or the initial stages of germination, including dipicolinic acid release. However, mutant spores exhibit a slight delay in the loss of optical density compared to that of wild-type spores. Mutants also retain more diaminopimelic acid and N-acetylmuramic acid during germination than wild-type spores, suggesting that the cortex peptidoglycan is not being hydrolyzed as rapidly. This finding is supported by high-pressure liquid chromatography analysis of the peptidoglycan structure used to confirm that SleL acts as an N-acetylglucosaminidase. When sleL is inactivated, the cortex peptidoglycan is not depolymerized into small muropeptides but instead is retained within the spore as large fragments. In the absence of the sleL-encoded N-acetylglucosaminidase, other cortex-lytic enzymes break down the cortex peptidoglycan sufficiently to allow rapid germination and outgrowth.
炭疽芽孢杆菌孢子是炭疽病的病原体,因其对多种根除方法具有抗性,所以从污染区域清除它们极为困难。这些抗性特性归因于孢子的脱水和休眠以及围绕孢子核心的多层保护层,其中一层是皮层。为使炭疽芽孢杆菌孢子萌发并恢复生长,皮层肽聚糖必须解聚。本研究报告了对编码一种皮层裂解酶的sleL(yaaH)的分析。sleL的失活不影响营养生长、孢子活力或萌发的初始阶段,包括吡啶二羧酸的释放。然而,与野生型孢子相比,突变体孢子在光密度损失方面表现出轻微延迟。在萌发过程中,突变体也比野生型孢子保留更多的二氨基庚二酸和N - 乙酰胞壁酸,这表明皮层肽聚糖没有被迅速水解。这一发现得到了用于确认SleL作为N - 乙酰葡糖胺酶的肽聚糖结构的高压液相色谱分析的支持。当sleL失活时,皮层肽聚糖不会解聚成小的胞壁肽,而是作为大的片段保留在孢子内。在没有sleL编码的N - 乙酰葡糖胺酶的情况下,其他皮层裂解酶会充分分解皮层肽聚糖,以允许快速萌发和生长。