The rate of obesity and the incidence of related diseases are rising at the same time. This is a serious threat to human health. Several studies have shown that reducing excessive intake of high-calorie sugars can reduce the risk of obesity, diabetes, hypertension and cancer! In 2015, the World Health Organization made a call to reduce sugar intake in children and adults. Sugar substitutes are a group of substances that have a sweet flavor but few or no calories compared to sucrose and glucose. They can provide sweetness while reducing calorie intake, which is of great significance for diabetics and people who need to lose weight. Among the natural sugar substitutes, erythritol and xylitol are two typical representatives. We constructed xylose reductase-expressing strains with different enzyme sources to catalyze xylose for xylitol production using fermentation with Escherichia coli (E. coli). The strategy of promoter engineering was utilized to improve the efficiency of conversion. We overexpressed the erythritol synthesis pathway enzyme in Y. lipolytica. Based on a high-throughput screening strategy, we constructed fluorescent reportor system for characterizing erythritol production and improving erythritol production in Y. lipolytica.