Deeble V J, Lindley H K, Fazeli M R, Cove J H, Baumberg S
Department of Genetics, University of Leeds, UK.
Microbiology (Reading). 1995 Oct;141 ( Pt 10):2511-8. doi: 10.1099/13500872-141-10-2511.
Streptomyces griseus ATCC 12475 fails to produce streptomycin when grown at 34 degrees C or above, although growth is appreciable up to at least 37 degrees C. This depression of streptomycin production at elevated growth temperature is manifest equally in liquid and on solid, and with complex and minimal, media. We report studies with gene fusions of the reporter genes aph or xyIE to restriction fragments containing the streptomycin biosynthesis promoter PstrB1. aph constructs were in high, and xyIE constructs in low, copy number vectors. Two strB1 promoter fragments were used, one requiring activation by the pathway-specific activator StrR of S. griseus, the other reportedly activator independent. PstrB1 expression in the aph constructs in S. griseus and in S. lividans was significantly reduced at 37 degrees C compared to 30 degrees C. Some of this reduction could be explained by lower plasmid copy number at the higher temperature, but strR-dependent expression was clearly temperature controlled. Using the xyIE reporter system, the temperature dependence of PstrB1 expression was confirmed but, surprisingly, the strR dependence of the two promoter fragments differed from that observed in the multicopy aph constructs. These data identify a temperature-dependent promoter which may contribute to the depressive effect of elevated growth temperature on streptomycin production.
灰色链霉菌ATCC 12475在34摄氏度或以上温度下生长时不产生链霉素,尽管在至少37摄氏度时生长明显。生长温度升高时链霉素产量的这种降低在液体和固体培养基以及复杂和基本培养基中均有体现。我们报告了将报告基因aph或xyIE与包含链霉素生物合成启动子PstrB1的限制片段进行基因融合的研究。aph构建体存在于高拷贝数载体中,xyIE构建体存在于低拷贝数载体中。使用了两个strB1启动子片段,一个需要灰色链霉菌的途径特异性激活剂StrR激活,另一个据报道不依赖激活剂。与30摄氏度相比,灰色链霉菌和变铅青链霉菌中aph构建体的PstrB1表达在37摄氏度时显著降低。这种降低部分可归因于较高温度下较低的质粒拷贝数,但StrR依赖性表达显然受温度控制。使用xyIE报告系统,证实了PstrB1表达的温度依赖性,但令人惊讶的是,两个启动子片段的StrR依赖性与在多拷贝aph构建体中观察到的不同。这些数据确定了一个温度依赖性启动子,它可能导致生长温度升高对链霉素生产的抑制作用。