Department of Veterinary Disease Biology, Faculty of Life Sciences, University of Copenhagen, Frederiksberg C, Denmark.
Appl Environ Microbiol. 2010 Oct;76(19):6470-6. doi: 10.1128/AEM.00297-10. Epub 2010 Jul 30.
Chitin, an insoluble polymer of N-acetyl-D-glucosamine (GlcNAc), is one of the most abundant carbohydrate polymers in marine and terrestrial environments. Chitin hydrolysis by Listeria monocytogenes depends on two chitinase-encoding genes, chiA and chiB, and the aim of this study was to investigate their regulation. Chitin induces the expression of both chitinases in late exponential growth phase, and chiA but not chiB is furthermore induced by the monomer GlcNAc. Furthermore, their expression is subjected to catabolite control. Chitinases expressed by bacterial pathogens have proven to be important not only for nutrient acquisition and environmental survival but also for infecting animals and humans. Interestingly, the central L. monocytogenes virulence gene regulator, PrfA, is required for the chitinolytic phenotype, as chitinase activity was significantly reduced in prfA mutant cells compared to its level in wild-type cells. In agreement with this, Northern blot analysis showed that the amounts of chiA and chiB transcripts upon induction by chitin were significantly lower in the prfA mutant than in the wild type. The chitinolytic activity and chiA and chiB expression were reduced in the absence of the sigB gene, indicating that σ(B) is also important for the production of chitinases. The chiA, chiB, and chiA chiB mutants were not impaired for in vitro adhesion and invasion in epithelial cell lines, but the chiA chiB double mutant showed less survival ability in a chitin-enriched medium. The regulation of chitinolytic activity in L. monocytogenes is complex, and taken together, the results indicate that the biological role of this activity may not be limited to the external environment.
几丁质是一种不溶于水的 N-乙酰-D-氨基葡萄糖(GlcNAc)聚合物,是海洋和陆地环境中最丰富的碳水化合物聚合物之一。单核细胞增生李斯特菌通过两种几丁质酶编码基因 chiA 和 chiB 水解几丁质,本研究旨在研究它们的调控机制。几丁质诱导迟滞期后期两种几丁质酶的表达,而单体 GlcNAc 进一步诱导 chiA 但不诱导 chiB 的表达。此外,它们的表达受到分解代谢物的控制。细菌病原体表达的几丁质酶不仅对营养物质的获取和环境生存至关重要,而且对感染动物和人类也很重要。有趣的是,李斯特菌的中心毒力基因调控因子 PrfA 对于几丁质酶表型是必需的,因为与野生型细胞相比,突变细胞中的几丁质酶活性显著降低。与这一结果一致的是,Northern 印迹分析显示,在几丁质诱导下 chiA 和 chiB 转录物的量在突变细胞中显著低于野生型细胞。在 sigB 基因缺失的情况下,几丁质酶活性和 chiA 和 chiB 的表达降低,表明 σ(B) 对于几丁质酶的产生也很重要。chiA、chiB 和 chiA chiB 突变体在体外黏附和上皮细胞系侵袭方面没有受损,但 chiA chiB 双突变体在富含几丁质的培养基中生存能力较低。李斯特菌中几丁质酶活性的调控很复杂,综合结果表明,这种活性的生物学作用可能不仅局限于外部环境。