Kelkar H S, Skloss T W, Haw J F, Keller N P, Adams T H
Department of Biology, Texas A&M University, College Station, Texas 77843, USA.
J Biol Chem. 1997 Jan 17;272(3):1589-94. doi: 10.1074/jbc.272.3.1589.
The Aspergillus nidulans stcL gene is predicted to encode a cytochrome P-450 monooxygenase and is located within a cluster of other genes that are required for synthesis of sterigmatocystin. Inactivation of stcL resulted in strains that accumulate dihydrosterigmatocystin, a tetrahydrobisfuran containing molecule that is very similar to the unsaturated product of the wild-type pathway, sterigmatocystin. This observation led us to hypothesize that the A. nidulans sterigmatocystin biosynthetic pathway is branched similarly to the aflatoxin pathway in Aspergillus parasiticus and Aspergillus flavus and that StcL is required for the desaturation of the bisfuran moiety in the sterigmatocystin/aflatoxin precursor versicolorin B. This prediction was confirmed by feeding the stcL mutant with the subsequent pathway intermediate, versicolorin A, which resulted in accumulation of both sterigmatocystin and dihydrosterigmatocystin, indicating that StcL functions before versicolorin A synthesis. A. nidulans stcU was shown previously to encode a ketoreductase required to convert versicolorin A to demethylsterigmatocystin and an stcL, stcU double mutant strain was shown here to accumulate only versicolorin B. These results indicate that both versicolorin A and versicolorin B can serve as substrates for StcU, resulting in a branched pathway. The final product of each branch are sterigmatocystin and dihydrosterigmatocystin, respectively.
构巢曲霉的stcL基因预计编码一种细胞色素P - 450单加氧酶,它位于与柄曲霉素合成所需的其他基因的一个簇内。stcL的失活导致菌株积累二氢柄曲霉素,这是一种含有四氢双呋喃的分子,与野生型途径的不饱和产物柄曲霉素非常相似。这一观察结果使我们推测,构巢曲霉的柄曲霉素生物合成途径与寄生曲霉和黄曲霉中的黄曲霉毒素途径类似地分支,并且StcL是柄曲霉素/黄曲霉毒素前体杂色曲霉素B中双呋喃部分去饱和所必需的。通过给stcL突变体提供后续途径中间体杂色曲霉素A来证实这一预测,这导致了柄曲霉素和二氢柄曲霉素的积累,表明StcL在杂色曲霉素A合成之前起作用。先前已表明构巢曲霉的stcU编码将杂色曲霉素A转化为去甲基柄曲霉素所需的酮还原酶,并且在此显示stcL、stcU双突变体菌株仅积累杂色曲霉素B。这些结果表明杂色曲霉素A和杂色曲霉素B都可以作为StcU的底物,从而导致一条分支途径。每个分支的最终产物分别是柄曲霉素和二氢柄曲霉素。