Analysis of two distinct mycelial populations in liquid-grown Streptomyces cultures using a flow cytometry-based proteomics approach
Analysis of two distinct mycelial populations in liquid-grown Streptomyces cultures using a flow cytometry-based proteomics approach
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DOI：
10.1007/s00253-012-4490-5
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发表时间：
2012-12-01
影响因子：
5
通讯作者：
Claessen, Dennis
中科院分区：
文献类型：
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作者：
van Veluw, G. Jerre;Petrus, Marloes L. C.;Claessen, Dennis
链霉菌是酶和抗生素的熟练生产者。当在生物反应器中生长时，这些丝状微生物形成由相互连接的菌丝组成的菌丝球。我们在这里采用了流式细胞术的方法设计的大颗粒（COPAS），并证明，液体生长的链霉菌文化由两个不同的群体的小球。一个群体由具有约260 μ m的恒定平均直径的菌丝体组成，而另一个群体包含较大的菌丝体，其直径取决于菌株、培养物的年龄和培养基组成。定量蛋白质组学分析表明，37个蛋白质的丰度不同的两个群体的颗粒。压力相关的蛋白质和生物合成蛋白质的钙依赖性抗生素的生产更丰富的人口中的大型菌丝体，而蛋白质参与DNA拓扑结构，修改，或降解的人口中的小菌丝体。纤维素酶样蛋白CslA和卓别林基因的缺失影响了大颗粒群体的平均大小，但不影响小颗粒的平均大小。考虑到酶和代谢产物的生产取决于颗粒大小的事实，这些结果提供了新的线索，走向合理的菌株设计的链霉菌株量身定制的工业发酵。
Streptomycetes are proficient producers of enzymes and antibiotics. When grown in bioreactors, these filamentous microorganisms form mycelial pellets that consist of interconnected hyphae. We here employed a flow cytometry approach designed for large particles (COPAS) and demonstrate that liquid-grown Streptomyces cultures consist of two distinct populations of pellets. One population consists of mycelia with a constant mean diameter of approximately 260 mu m, whereas the other population contains larger mycelia whose diameter depends on the strain, the age of the culture, and medium composition. Quantitative proteomics analysis revealed that 37 proteins differed in abundance between the two populations of pellets. Stress-related proteins and biosynthetic proteins for production of the calcium-dependent antibiotic were more abundant in the population of large mycelia, while proteins involved in DNA topology, modification, or degradation were overrepresented in the population of small mycelia. Deletion of genes for the cellulose synthase-like protein CslA and the chaplins affected the average size of the population of large pellets but not that of small pellets. Considering the fact that the production of enzymes and metabolites depends on pellet size, these results provide new leads toward rational strain design of Streptomyces strains tailored for industrial fermentations.