Abstract
It has been reported that trehalose plays an important role in stress tolerance in yeasts. Therefore, in order to construct a stably recombinant Saccharomyces sp. W0 with higher ethanol tolerance, the TPS1 gene encoding 6-phosphate-trehalose synthase cloned from Saccharomycopsis fibuligera A11 was ligated into the 18S rDNA integration vector pMIRSC11 and integrated into chromosomal DNA of Saccharomyces sp. W0. The transformant Z8 obtained had the content of 6.23g of trehalose/100g of cell dry weight, while Saccharomyces sp. W0 only contained 4.05g of trehalose/100g of cell dry weight. The transformant Z8 also had higher ethanol tolerance (cell survival was 25.1% at 18ml of ethanol/100ml of solution) and trehalose-6-phosphate synthase (Tps1) activity (1.3U/mg) and produced more ethanol (16.4ml of ethanol/100ml of medium) than Saccharomyces sp. W0 (cell survival was 12.1% at 18ml of ethanol/100ml of solution, Tps1 activity was 0.8U/mg and the produced ethanol concentration was 14.2ml of ethanol/100ml of medium) under the same conditions. The results show that trehalose indeed can play an important role in ethanol tolerance and ethanol production by Saccharomyces sp. W0.
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