Abstract

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.

Key points

• We described a design-build-test-learn pipeline to construct in vitro biosystems.

• The designed system comprised six key enzymes and another three enzymes.

• The system converted maltodextrin stoichiometrically to mannitol in one pot.

中文翻译：

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不含ATP的体外合成酶生物系统，促进淀粉一锅化学计量转化为甘露醇

摘要

D-甘露醇（以下称为甘露醇）是六碳糖醇，在食品和制药工业中具有多种应用。为了克服目前通常用于甘露醇生产的化学氢化方法的缺点，需要寻找高收率和低成本的新型预期的甘露醇生产策略。在这项研究中，我们提出了一种新颖的方法，用于通过使用低成本淀粉作为底物的体外合成酶生物系统化学计量合成甘露醇。通过将整个反应路径分为三个模块，这些模块可以在一个罐中依次执行，我们的设计旨在以不依赖ATP和辅因子的方式将淀粉基材料化学计量转化为甘露醇。在最佳条件下，使用10 g / L和50 g / L的麦芽糖糊精作为底物，可实现约95-98％的高产品收率，这表明我们设计的系统具有工业应用潜力。这项研究不仅为合成甘露醇提供了一种高效的策略，而且通过使用低成本淀粉基材料作为输入的体外合成酶生物系统扩大了糖醇的产品范围。

关键点

•我们描述了构建体外生物系统的设计-构建-测试-学习管道。

•设计的系统包含六个关键酶和另外三个酶。

•该系统在一锅中将麦芽糖糊精化学计量地转化为甘露醇。