TY - JOUR
T1 - Structural insights into the Smirnoff-Wheeler pathway for vitamin C production in the Amazon fruit camu-camu
AU - Vargas, Jhon A.
AU - Sculaccio, Susana A.
AU - Pinto, Andressa P.A.
AU - Pereira, Humberto D.Muniz
AU - Mendes, Luis F.S.
AU - Flores, Jhoao F.
AU - Cobos, Marianela
AU - Castro, Juan C.
AU - Garratt, Richard C.
AU - Leonardo, Diego A.
N1 - Publisher Copyright:
© 2024 The Author(s).
PY - 2024/5/3
Y1 - 2024/5/3
N2 - l-Ascorbic acid (AsA, vitamin C) is a pivotal dietary nutrient with multifaceted importance in living organisms. In plants, the Smirnoff-Wheeler pathway is the primary route for AsA biosynthesis, and understanding the mechanistic details behind its component enzymes has implications for plant biology, nutritional science, and biotechnology. As part of an initiative to determine the structures of all six core enzymes of the pathway, the present study focuses on three of them in the model species Myrciaria dubia (camu-camu): GDP-d-mannose 3',5'-epimerase (GME), l-galactose dehydrogenase (l-GalDH), and l-galactono-1,4-lactone dehydrogenase (l-GalLDH). We provide insights into substrate and cofactor binding and the conformational changes they induce. The MdGME structure reveals a distorted substrate in the active site, pertinent to the catalytic mechanism. Mdl-GalDH shows that the way in which NAD+ association affects loop structure over the active site is not conserved when compared with its homologue in spinach. Finally, the structure of Mdl-GalLDH is described for the first time. This allows for the rationalization of previously identified residues which play important roles in the active site or in the formation of the covalent bond with FAD. In conclusion, this study enhances our understanding of AsA biosynthesis in plants, and the information provided should prove useful for biotechnological applications.
AB - l-Ascorbic acid (AsA, vitamin C) is a pivotal dietary nutrient with multifaceted importance in living organisms. In plants, the Smirnoff-Wheeler pathway is the primary route for AsA biosynthesis, and understanding the mechanistic details behind its component enzymes has implications for plant biology, nutritional science, and biotechnology. As part of an initiative to determine the structures of all six core enzymes of the pathway, the present study focuses on three of them in the model species Myrciaria dubia (camu-camu): GDP-d-mannose 3',5'-epimerase (GME), l-galactose dehydrogenase (l-GalDH), and l-galactono-1,4-lactone dehydrogenase (l-GalLDH). We provide insights into substrate and cofactor binding and the conformational changes they induce. The MdGME structure reveals a distorted substrate in the active site, pertinent to the catalytic mechanism. Mdl-GalDH shows that the way in which NAD+ association affects loop structure over the active site is not conserved when compared with its homologue in spinach. Finally, the structure of Mdl-GalLDH is described for the first time. This allows for the rationalization of previously identified residues which play important roles in the active site or in the formation of the covalent bond with FAD. In conclusion, this study enhances our understanding of AsA biosynthesis in plants, and the information provided should prove useful for biotechnological applications.
KW - Camu-camu
KW - eEnzyme structure
KW - GDP-d-mannose 3',5'-epimerase
KW - L-Ascorbic acid
KW - L-galactono-1,4-lactone dehydrogenase Myrciaria dubia
KW - L-galactose dehydrogenase
KW - Smirnoff-Wheeler pathway
KW - Vitamin C
UR - http://www.scopus.com/inward/record.url?scp=85192849948&partnerID=8YFLogxK
U2 - 10.1093/jxb/erae016
DO - 10.1093/jxb/erae016
M3 - Artículo
AN - SCOPUS:85192849948
SN - 0022-0957
VL - 75
SP - 2754
EP - 2771
JO - Journal of Experimental Botany
JF - Journal of Experimental Botany
IS - 9
ER -