Trudelle Danielle M, Bryan Daniel W, Ray Shahla, Munafo John P, Denes Thomas G
Department of Food Science, University of Tennessee, Knoxville, Tennessee 37996, United States.
ACS Omega. 2022 May 11;7(20):17002-17013. doi: 10.1021/acsomega.1c07403. eCollection 2022 May 24.
is a Gram-positive foodborne pathogen that causes listeriosis, an illness that may result in serious health consequences or death. Wall teichoic acids (WTAs) are external cell wall glycopolymers that play many biological roles. Here, the WTA composition was determined for several phage-resistant mutant strains of . The strains included wild-type (WT) 10403S, and three phage-resistant mutant strains derived from 10403S, consisting of two well-characterized strains and one with unknown impact on cell physiology. Several WTA monomers were prepared from WT 10403S, as analytical standards. The WTA monomer fraction was then isolated from the mutant strains and the corresponding per-trimethylsilylated derivatives were analyzed by gas chromatography-flame ionization detection. WTA monomer, GlcNAc-Rha-Rbo, was detected in 10403S, and not detected in the phage-resistant strains known to lack rhamnose and -acetylglucosamine; although the expected monomers GlcNAc-Rbo and Rha-Rbo were detected, respectively. GlcNAc-Rha-Rbo was also detected in strain UTK P1-0001, which is known to impact phage adsorption through an undetermined mechanism, albeit at a lower intensity than the WT 10403S, which is consistent with partial loss of function through truncation in RmlC protein. WTA monomers were also detected in an unpurified cell pellet, demonstrating that the method employed in this study can be used to rapidly screen without laborious WTA purification. This study lays the groundwork for future studies on WTA compositional analysis to support genomic data, and serves as a foundation for the development of new rapid methods for WTA compositional analysis.
是一种革兰氏阳性食源性病原体，可导致李斯特菌病，这种疾病可能会导致严重的健康后果甚至死亡。壁磷壁酸（WTA）是外部细胞壁糖聚合物，发挥着多种生物学作用。在此，测定了几种 的噬菌体抗性突变菌株的WTA组成。这些菌株包括野生型（WT）10403S，以及源自10403S的三种噬菌体抗性突变菌株，其中包括两种特征明确的菌株和一种对细胞生理学影响未知的菌株。从WT 10403S制备了几种WTA单体作为分析标准品。然后从突变菌株中分离出WTA单体部分，并通过气相色谱 - 火焰离子化检测分析相应的全三甲基硅烷基化衍生物。在10403S中检测到WTA单体GlcNAc - Rha - Rbo，而在已知缺乏鼠李糖和N - 乙酰葡糖胺的噬菌体抗性菌株中未检测到；尽管分别检测到了预期的单体GlcNAc - Rbo和Rha - Rbo。在菌株UTK P1 - 0001中也检测到了GlcNAc - Rha - Rbo，已知该菌株通过一种未确定的机制影响噬菌体吸附，尽管其强度低于WT 10403S，这与RmlC蛋白截短导致的部分功能丧失一致。在未纯化的细胞沉淀中也检测到了WTA单体，这表明本研究中采用的方法可用于快速筛选，而无需费力地纯化WTA。本研究为未来支持基因组数据的WTA组成分析研究奠定了基础，并为开发新的WTA组成分析快速方法提供了基础。
