Tani Shuji, Yuki Shota, Kunitake Emi, Sumitani Jun-Ichi, Kawaguchi Takashi
a Graduate School of Life and Environmental Sciences , Osaka Prefecture University , Sakai , Japan.
Biosci Biotechnol Biochem. 2017 Jun;81(6):1227-1234. doi: 10.1080/09168451.2017.1295800. Epub 2017 Mar 14.
We screened for factors involved in the cellulose-responsive induction of cellulose biomass-degrading enzyme genes from approximately 12,000 Aspergillus aculeatus T-DNA insertion mutants harboring a transcriptional fusion between the FIII-avicelase gene (cbhI) promoter and the orotidine 5'-monophosphate decarboxylase gene. Analysis of 5-fluoroorodic acid (5-FOA) sensitivity, cellulose utilization, and cbhI expression of the mutants revealed that a mutant harboring T-DNA at the dipeptidyl peptidase IV (dppIV) locus had acquired 5-FOA resistance and was deficient in cellulose utilization and cbhI expression. The deletion of dppIV resulted in a significant reduction in the cellulose-responsive expression of both cbhI as well as genes controlled by XlnR-independent and XlnR-dependent signaling pathways at an early phase in A. aculeatus. In contrast, the dppIV deletion did not affect the xylose-responsive expression of genes under the control of XlnR. These results demonstrate that DppIV participates in cellulose-responsive induction in A. aculeatus.
我们从大约12,000个棘孢曲霉T-DNA插入突变体中筛选了参与纤维素响应诱导纤维素生物质降解酶基因的因子,这些突变体含有III型葡聚糖内切酶基因(cbhI)启动子与乳清苷5'-单磷酸脱羧酶基因之间的转录融合。对突变体的5-氟乳清酸(5-FOA)敏感性、纤维素利用和cbhI表达的分析表明,在二肽基肽酶IV(dppIV)位点携带T-DNA的突变体获得了5-FOA抗性,并且在纤维素利用和cbhI表达方面存在缺陷。dppIV的缺失导致在棘孢曲霉早期阶段,cbhI以及由XlnR非依赖性和XlnR依赖性信号通路控制的基因的纤维素响应表达显著降低。相反,dppIV缺失不影响XlnR控制下的基因的木糖响应表达。这些结果表明DppIV参与了棘孢曲霉中的纤维素响应诱导。