Measurement of lactose concentration in milk by using engineered bacteria producing lycopene

Lycopene is an orange-red carotenoid, which confers a visual phenotype to assess genetic transformation of microorganisms. In this study, the lycopene synthesis pathway was constructed in engineered Escherichia coli BL21 (DE3) by transforming plasmid pET-15b-crtBEI, wherein crtB, crtE, and crtI could be expressed under the control of the T7 promoter and lacO operator and lycopene could be accumulated in the engineered bacteria upon induction by lactose. A good linear relationship was observed between the lycopene content in engineered bacterial culture and lactose concentration in the range of 4-52 g/L; using this relation, the lactose concentration in milk could be determined. This method could be used to overcome several limitations of the high-performance liquid chromatography (HPLC) method for lactose detection, such as cumbersome sample preparation and expensive detection equipment. Moreover, this method required only a clean bench, shaker, and spectrophotometer for lactose analysis. Additionally, no significant difference was observed between this method and HPLC in terms of lactose measurement in milk, indicating that this method is reasonable and simple.