Hughes Lee E, Beck Debrah A, O'Donovan Gerard A
Department of Biological Sciences, University of North Texas, PO 305220, Denton, TX 76203, USA.
Curr Microbiol. 2005 Jan;50(1):8-10. doi: 10.1007/s00284-004-4386-4. Epub 2004 Dec 8.
Using 5-fluoropyrimidine analogues, high-performance liquid chromatography (HPLC), and the feeding of pyrimidine compounds to pyrimidine auxotrophs, the pathways for salvage of exogenous pyrimidine nucleosides and bases in Streptomyces were established. Selection for resistance to the analogues resulted in the isolation of strains of S. griseus lacking the following enzyme activities: uracil phosphoribosyltransferase (upp) and cytidine deaminase (cdd). The conversion of substrates in the pathway was followed using reverse-phase HPLC. The strains deficient in salvage enzymes were also verified by this method. In addition, feeding of exogenous pyrimidines to strains lacking the biosynthetic pathway confirmed the salvage pathway. Data from the analogue, HPLC, and feeding experiments showed that Streptomyces recycles the pyrimidine base uracil, as well as the nucleosides uridine and cytidine. Cytosine is not recycled due to a lack of cytosine deaminase.
利用5-氟嘧啶类似物、高效液相色谱法(HPLC)以及向嘧啶营养缺陷型菌株投喂嘧啶化合物,确定了链霉菌中外源嘧啶核苷和碱基的补救途径。通过选择对类似物的抗性,分离出了缺乏以下酶活性的灰色链霉菌菌株:尿嘧啶磷酸核糖基转移酶(upp)和胞苷脱氨酶(cdd)。使用反相HPLC追踪该途径中底物的转化。缺乏补救酶的菌株也通过该方法得到了验证。此外,向缺乏生物合成途径的菌株投喂外源嘧啶证实了补救途径。来自类似物、HPLC和投喂实验的数据表明,链霉菌可循环利用嘧啶碱基尿嘧啶以及核苷尿苷和胞苷。由于缺乏胞嘧啶脱氨酶,胞嘧啶无法循环利用。