Molecular basis for branched steviol glucoside biosynthesis

Lee, Soon Goo; Salomon, Eitan; Yu, Oliver; Jez, Joseph M.

doi:10.1073/pnas.1902104116

Title: Molecular basis for branched steviol glucoside biosynthesis

Journal Article · 10 June 2019 · Proceedings of the National Academy of Sciences of the United States of America

DOI: https://doi.org/10.1073/pnas.1902104116 · OSTI ID:1545888

Lee, Soon Goo ^[1]; Salomon, Eitan ^[2]; Yu, Oliver ^[3]; Jez, Joseph M. ^[4]

Washington Univ., St. Louis, MO (United States); North Carolina State Univ., Wilmington, NC (United States)
Washington Univ., St. Louis, MO (United States); National Center for Mariculture, Israel Oceanographic and Limnological Research, Eilat (Israel)
Conagen, Inc., Bedford, MA (United States)
Washington Univ., St. Louis, MO (United States)

Steviol glucosides, such as stevioside and rebaudioside A, are natural products roughly 200-fold sweeter than sugar and are used as natural, noncaloric sweeteners. Biosynthesis of rebaudioside A, and other related stevia glucosides, involves formation of the steviol diterpenoid followed by a series of glycosylations catalyzed by uridine diphosphate (UDP)-dependent glucosyltransferases. UGT76G1 from Stevia rebaudiana catalyzes the formation of the branched-chain glucoside that defines the stevia molecule and is critical for its high-intensity sweetness. Here, we report the 3D structure of the UDP-glucosyltransferase UGT76G1, including a complex of the protein with UDP and rebaudioside A bound in the active site. The X-ray crystal structure and biochemical analysis of site-directed mutants identifies a catalytic histidine and how the acceptor site of UGT76G1 achieves regioselectivity for branched-glucoside synthesis. The active site accommodates a two-glucosyl side chain and provides a site for addition of a third sugar molecule to the C3' position of the first C13 sugar group of stevioside. This structure provides insight on the glycosylation of other naturally occurring sweeteners, such as the mogrosides from monk fruit, and a possible template for engineering of steviol biosynthesis.

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Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES)

Grant/Contract Number:: AC02-06CH11357

OSTI ID:: 1545888

Journal Information:: Proceedings of the National Academy of Sciences of the United States of America, Vol. 116, Issue 26; ISSN 0027-8424

Publisher:: National Academy of SciencesCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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Cited by: 35 works

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59 BASIC BIOLOGICAL SCIENCES
glucosyltransferase
noncaloric sweetener
plant biochemistry
stevia
X-ray crystal structure

Title: Molecular basis for branched steviol glucoside biosynthesis

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