18S rDNA integration of the exo-inulinase gene into chromosomes of the high ethanol producing yeast Saccharomyces sp. W0 for direct conversion of inulin to bioethanol

Inulin and inulin-containing materials are good substrates for bioethanol production. In order to construct genetically stable recombinant yeast cells carrying the inulinase gene for bioethanol production from inulin, a new 18S rDNA integration vector was constructed in this study. After the INU1 gene encoding exo-inulinase was ligated into the 18S rDNA integration vector, transformed into uracil mutant of Saccharomyces sp. W0 and integrated into its chromosomes, the transformant MguINU1-04 obtained could produce 34.8 U cm−3 of inulinase activity within 72 h. The transformant could stably produce high activity of inulinase after five sequential batch cultivations. During 5-l fermentation, ethanol concentration in the fermentation medium containing 30.0% inulin was 14.7% (v/v) and the ethanol productivity was over 0.386 g of ethanol per g of inulin. When the tuber meal of Jerusalem artichoke (50.0%) was fermented into ethanol by the transformant MguINU1-04, 12.6% (v/v) ethanol was produced within 120 h and the ethanol productivity was over 0.331 g of ethanol per g of sugar. Therefore, inulin and inulin in the tuber meal of Jerusalem artichoke could be directly converted into high concentrations of ethanol by the engineered Saccharomyces sp. W0 carrying the inulinase gene.

► It was found that after the INU1 gene was inserted into chromosomes of Saccharomyces sp. W0 via the 18S rDNA integration vector, the transformant MguINU1-04 obtained could stably produce around 34.8 U/ml of inulinase activity within 72 h.
► During 5-l fermentation, ethanol concentration in the fermented medium containing 30% (w/v) inulin was 14.7% (v/v).
► When the tuber meal of Jerusalem artichoke (50.0% w/v) was fermented into ethanol by the transformant MguINU1-04, 12.6% (v/v).