Production of steviol glycosides through whole cell biotransformation of steviol glycoside intermediates

A method for producing a steviol glycoside product from steviol glycoside intermediates, comprising:

• providing a bacterial cell expressing one or more UDP-dependent glycosyl transferase enzymes (UGT enzymes) intracellularly, wherein the UGT enzymes glycosylate steviol and steviol glycoside substrates, and wherein the bacterial cell comprises the following genetic modifications:
• deletion, inactivation, or reduced activity or expression of UDP-sugar hydrolase and one or more UDP-galactose biosynthesis enzymes;
• deletion, inactivation, or reduced activity or expression of glucose-6-phosphate isomerase;
• and
• overexpression of phosphoglucomutase and UTP-glucose-1-phosphate uridylyltransferase;
• incubating the bacterial cell with a stevia leaf extract or fraction thereof comprising the steviol glycoside intermediates thereby glycosylating the steviol glycoside intermediates to the steviol glycoside product by enzymatic transfer of one or more glucosyl groups from UDP-glucose cofactor, and
• recovering the steviol glycoside product.