4.7 Article

An ATP-free in vitro synthetic enzymatic biosystem facilitating one-pot stoichiometric conversion of starch to mannitol

Journal

APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Volume 105, Issue 5, Pages 1913-1924

Publisher

SPRINGER
DOI: 10.1007/s00253-021-11154-9

Keywords

D-Mannitol; Maltodextrin; Cascade biocatalysis; In vitro synthetic biology; Reaction modules

Funding

  1. National Natural Science Foundation of China [21778073]
  2. Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project [TSBICIP-KJGG-003]

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A novel approach for stoichiometric synthesis of mannitol through an in vitro synthetic enzymatic biosystem using low-cost starch as substrate is proposed in this study. The overall reaction pathway is divided into three modules which could be executed sequentially in one pot. High product yields of around 95-98% were achieved under optimized conditions.
D-Mannitol (hereinafter as mannitol) is a six-carbon sugar alcohol with diverse applications in food and pharmaceutical industries. To overcome the drawbacks of the chemical hydrogenation method commonly used for mannitol production at present, there is a need to search for novel prospective mannitol production strategies that are of high yield and low cost. In this study, we present a novel approach for the stoichiometric synthesis of mannitol via an in vitro synthetic enzymatic biosystem using the low-cost starch as substrate. By dividing the overall reaction pathway into three modules which could be executed sequentially in one pot, our design aimed at the stoichiometric conversion of starch-based materials into mannitol in an ATP-independent and cofactor-balanced manner. At optimized conditions, high product yields of around 95-98% were achieved using both 10 g/L and 50 g/L maltodextrin as substrate, indicating the potential of our designed system for industrial applications. This study not only provides a high-efficient strategy for the synthesis of mannitol but also expands the product scope of sugar alcohols by the in vitro synthetic enzymatic biosystems using low-cost starch-based materials as the input.

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