The role of cytosolic alpha-glucan phosphorylase in maltose metabolism and the comparison of amylomaltase in Arabidopsis and Escherichia coli.
Transitory starch of leaves is broken down hydrolytically, making maltose the predominant form of carbon exported from chloroplasts at night. Maltose metabolism in the cytoplasm of Escherichia coli requires amylomaltase (MalQ) and maltodextrin phosphorylase (MalP). Possible orthologs of MalQ and MalP in the cytosol of Arabidopsis (Arabidopsis thaliana) were proposed as disproportionating enzyme (DPE2, At2g40840) and alpha-glucan phosphorylase (AtPHS2, At3g46970). In this article, we measured the activities of recombinant DPE2 and AtPHS2 proteins with various substrates; we show that maltose and a highly branched, soluble heteroglycan (SHG) are excellent substrates for DPE2 and propose that a SHG is the in vivo substrate for DPE2 and AtPHS2. In E. coli, MalQ and MalP preferentially use smaller maltodextrins (G(3)-G(7)) and we suggest that MalQ and DPE2 have similar, but nonidentical, roles in maltose metabolism. To study this, we complemented a MalQ(-) E. coli strain with DPE2 and found that the rescue was not complete. To investigate the role of AtPHS2 in maltose metabolism, we characterized a T-DNA insertion line of the AtPHS2 gene. The nighttime maltose level increased 4 times in the Atphs2-1 mutant. The comparison of maltose metabolism in Arabidopsis with that in E. coli and the comparison of the maltose level in plants lacking DPE2 or AtPHS2 indicate that an alternative route to metabolize the glucan residues in SHG exists. Other plant species also contain SHG, DPE2, and alpha-glucan phosphorylase, so this pathway for maltose metabolism may be widespread among plants.
Lu
Yan
Y
Department of Botany , University of Wisconsin, Madison, Wisconsin 53706, USA.
Steichen
Jon M
JM
Yao
Jian
J
Sharkey
Thomas D
TD
D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics; D002940:Circadian Rhythm; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D005964:Glucosyltransferases / Q000235:genetics / Q000378:metabolism*; D006004:Glycogen Debranching Enzyme System / Q000378:metabolism*; D008320:Maltose / Q000378:metabolism*; D008969:Molecular Sequence Data; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism; D006005:Phosphorylases / Q000378:metabolism*; D013379:Substrate Specificity
D006004:Glycogen Debranching Enzyme System; D010750:Phosphoproteins; D008320:Maltose; D005964:Glucosyltransferases; D006005:Phosphorylases; C033938:maltodextrin phosphorylase; C020569:4 alpha-glucanotransferase