Low-alcohol Huangjiu (LAH), which contains reduced contents of ethanol and higher alcohols, is prepared by diluting original Huangjiu that has a high ethanol content, which leads to a weakened flavor (i.e., acidity). To increase acidity and reduce higher alcohols level in LAH, the gene ALD6 encoding aldehyde dehydrogenase was expressed in yeast HJ-1 under the control of the pPGK1 promoter and terminators with varying activities (tGIC1, tPGK1 and tCPS1) by scarless replacement at BAT2 locus, yielding the engineered strains HJΔB-AG, HJΔB-AP, and HJΔB-AC. The acetate concentration produced by HJΔB-AG, HJΔB-AP, and HJΔB-AC was 1.26-, 1.84-, and 2.51-fold of that of HJ-1, respectively. Furthermore, the concentration of higher alcohols produced by HJΔB-AG, HJΔB-AP, and HJΔB-AC decreased by 39.91%, 45.55%, and 52.80%, respectively. This study resulted in the creation of promising recombinant yeast strains and introduced a method that can be used for the high-quality production of LAH by acid-producing Saccharomyces cerevisiae.
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