	US 12,012,626 B2
	Production of steviol glycosides through whole cell biotransformation of steviol glycoside intermediates
Ajikumar Parayil Kumaran, Watertown, MA (US); Christine Nicole S. Santos, Newton, MA (US); Jason Eric Donald, Lexington, MA (US); Mary Elizabeth Fowler, Somerville, MA (US); Ryan N. Philippe, Somerville, MA (US); Christopher Scott Frei, Weymouth, MA (US); and Aaron Love, Boston, MA (US)
Assigned to Manus Bio Inc., Cambridge, MA (US)
Filed by Manus Bio Inc., Cambridge, MA (US)
Filed on Dec. 9, 2021, as Appl. No. 17/546,758.
Application 17/546,758 is a continuation of application No. 16/512,874, filed on Jul. 16, 2019, granted, now 11,230,724.
Claims priority of provisional application 62/698,617, filed on Jul. 16, 2018.
Prior Publication US 2022/0162658 A1, May 26, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12P 19/18 (2006.01)

CPC C12P 19/18 (2013.01)

17 Claims

1. A method for producing a glycosylated product from 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 glycoside substrates, wherein the bacterial cell has an overexpression of phosphoglucomutase (pgm) and UTP-glucose-1-phosphate uridylyltransferase; and a deletion, inactivation, or reduced expression of one or more genes encoding enzymes that consume UDP-glucose selected from a UDP-sugar hydrolase, an enzyme responsible for UDP-galactose biosynthesis from UDP-glucose, a trehalose-6-phosphate synthase, and a UDP-glucose-6-dehydrogenase,

incubating the bacterial strain with the glycoside intermediates, wherein the glycoside intermediates are steviol glycosides from stevia leaf extract, and

recovering the glycosylated product.