Räty K, Kunnari T, Hakala J, Mäntsälä P, Ylihonko K
Department of Biochemistry and Food Chemistry, Turku University, Finland.
Mol Gen Genet. 2000 Sep;264(1-2):164-72. doi: 10.1007/s004380000306.
We have cloned and characterized a gene cluster for anthracycline biosynthesis from Streptomyces galilaeus. This cluster, 15-kb long, includes eight genes involved in the deoxyhexose biosynthesis pathway, a gene for a glycosyltransferase and one for an activator, as well as two genes involved in aglycone biosynthesis. Gene disruption targeted to the activator gene blocked production of aclacinomycins in S. galilaeus. Plasmid pSgs4, containing genes for a glycosyltransferase (aknS), an aminomethylase (aknX), a glucose-1-phosphate thymidylyltransferase (akn Y) and two genes for unidentified glycosylation functions (aknT and aknV), restored the production of aclacinomycins in the S. galilaeus mutants H063, which accumulates aklavinone, and H054, which produces aklavinone with rhodinose and deoxyfucose residues. Furthermore, pSgs4 directed the production of L-rhamnosyl-epsilon-rhodomycinone and L-daunosaminyl-epsilon-rhodomycinone in S. peucetius strains that produce epsilon-rhodomycinone endogenously. Subcloning of the gene cluster was carried out in order to further define the genes that are responsible for complementation and hybrid anthracycline generation.
我们已经克隆并鉴定了来自加利利链霉菌的蒽环类抗生素生物合成基因簇。这个长15 kb的基因簇包括参与脱氧己糖生物合成途径的8个基因、一个糖基转移酶基因和一个激活剂基因，以及参与糖苷配基生物合成的两个基因。针对激活剂基因的基因破坏阻断了加利利链霉菌中阿克拉霉素的产生。质粒pSgs4含有糖基转移酶（aknS）、氨基甲基化酶（aknX）、葡萄糖-1-磷酸胸苷酰转移酶（aknY）的基因以及两个具有未知糖基化功能的基因（aknT和aknV），恢复了加利利链霉菌突变体H063（积累阿克拉维酮）和H054（产生带有鼠李糖和脱氧岩藻糖残基的阿克拉维酮）中阿克拉霉素的产生。此外，pSgs4指导了在能内生产生ε-红霉酮的产黄青霉菌株中L-鼠李糖基-ε-红霉酮和L-柔红糖胺基-ε-红霉酮的产生。为了进一步确定负责互补作用和杂合蒽环类抗生素产生的基因，进行了基因簇的亚克隆。
