Campus Units

Biochemistry, Biophysics and Molecular Biology, Roy J. Carver Department of

Document Type

Article

Publication Version

Published Version

Publication Date

11-25-2016

Journal or Book Title

Journal of Biological Chemistry

Volume

291

Issue

48

First Page

24951

Last Page

24960

DOI

10.1074/jbc.M116.754705

Abstract

A comprehensive description of starch biosynthesis and granule assembly remains undefined despite the central nature of starch as an energy storage molecule in plants and as a fundamental calorie source for many animals. Multiple theories regarding the starch synthase (SS)-catalyzed assembly of (α1–4)-linked D-glucose molecules into maltodextrins generally agree that elongation occurs at the non-reducing terminus based on the degradation of radiolabeled maltodextrins, although recent reports challenge this hypothesis. Surprisingly, a direct analysis of the SS catalytic product has not been reported, to our knowledge. We expressed and characterized recombinant Zea mays SSIIa and prepared pure ADP-[13CU]glucose in a one-pot enzymatic synthesis to address the polarity of maltodextrin chain elongation. We synthesized maltoheptaose (degree of polymerization 7) using ADP-[13CU]glucose, maltohexaose (degree of polymerization 6), and SSIIa. Product analysis by ESI-MS revealed that the [13CU]glucose unit was added to the non-reducing end of the growing chain, and SSIIa demonstrated a >7,850-fold preference for addition to the non-reducing end versus the reducing end. Independent analysis of [13CU]glucose added to maltohexaose by SSIIa using solution NMR spectroscopy confirmed the polarity of maltodextrin chain elongation.

Comments

This research was originally published in the Journal of Biological Chemistry. Larson, Mark E., Daniel J. Falconer, Alan M. Myers, and Adam W. Barb. "Direct characterization of the maize starch synthase IIa product shows maltodextrin elongation occurs at the non-reducing end." Journal of Biological Chemistry 291, no. 48 (2016): 24951-24960. © the American Society for Biochemistry and Molecular Biology. doi: 10.1074/jbc.M116.754705.

Copyright Owner

The American Society for Biochemistry and Molecular Biology, Inc.

Language

en

File Format

application/pdf

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