Abstract
Equol (EQ) and its analogous derivatives 5-hydroxy-equol (5-OH-EQ) and 5-hydroxy-dehydroequol (5-OH-D-EQ) are isoflavones which benefit human health. They are produced from daidzein and genistein, respectively, in the gut by microorganisms harboring the genes daidzein reductase (dzr), dihydrodaidzein racemase (ifcA), dihydrodaidzein reductase (ddr) and tetrahydrodaidzein reductase (tdr). Since the production of these isoflavones is of interest due to their great-health benefits for humans, the heterologous expression of dzr, ddr, tdr and ifcA from Slackia isoflavoniconvertenes DSM 22006T in lactic acid bacteria (LAB) was used as a strategy to produce EQ, 5-OH-EQ and 5-OH-D-EQ in soy beverages. However, efficient production of these compounds was only demonstrated in two engineered Limosilactobacillus fermentum strains, and it is dependent on dihydrodaidzein racemase (DDRC). In order to increase the production of EQ and its analogous derivatives in different LAB species and genera, different strategies were performed with the ifcA gene. Translational coupling of ifcA and dzr genes (pNZ:TuR.dzr.ifcA) under the influence of a constitutive promoter improved the efficiency of production of EQ, 5-OH-EQ and 5-OH-D-EQ in the engineered LAB strains. The translational coupling of ifcA and dzr genes allowed the production of high concentrations of eq (111.15 ± 9.20-410.56 ± 24.15 μM), 5-OH-eq (71.00 ± 4.25 μM-148.22 ± 9.15 μM) and 5-OH-D-eq (111.15 ± 9.20-201.09 ± 7.65 μM) in soy beverages by different engineered LAB genera, such as L. fermentum INIA 584L, Lactilactobacillus plantarum WCFS1, and Lactocaseibacillus paracasei BL23. Translational coupling has allowed engineered Laboratories strains belonging to different genera, such as L. fermentum, L. plantarum, and L. paracasei, to produce high concentrations of EQ, 5-OH-EQ and 5-OH-D-EQ. Translational coupling could be exploited as a strategy for the efficient production of bioactive compounds.