An in vitro synthetic biosystem based on acetate for production of phloroglucinol.
An in vitro synthetic biosystem based on acetate for production of phloroglucinol.
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用于生产间苯三酚的基于乙酸盐的体外合成生物系统
DOI：
10.1186/s12896-017-0376-z
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发表时间：
2017-08-08
影响因子：
3.5
通讯作者：
Liu H
中科院分区：
文献类型：
--
作者：
Zhang R;Liu W;Cao Y;Xu X;Xian M;Liu H
间苯三酚是一种重要的化学物质，目前已经建立了将葡萄糖转化为间苯三酚的生物合成工艺。然而，由于大约80%的葡萄糖被转化为不需要的副产物和生物质，这种生物合成过程仅显示出较低的产量，最高约为0.20 g/g。其工业应用通常受到低电流生产率和成品率以及高成本的阻碍。一般来说，几个不同的方面限制了间苯三酚生物合成的发展。间苯三酚的产率是其生物转化最重要的参数之一，特别是从经济和生态的角度来看。生物基化学物质的体外生物合成是一种灵活的、具有潜在高产率的替代体内生物合成技术。结果通过比较大肠杆菌和巴氏醋酸杆菌乙酰辅酶a合成酶（ACS）的活性，鉴定出高活性的ACS2。pasteurianus。表达并纯化了产自钙动杆菌的乙酰辅酶a羧化酶（ACC）和产自荧光假单胞菌的间苯三酚合成酶（PhlD）。通过上述酶和所需辅因子的联合作用，乙酸酯成功转化为间苯三酚。对体外反应体系进行优化后，间苯三酚的产率接近0.64 g /g乙酸，为理论最大值的91.43%。结论建立了一种以乙酸酯为原料高效制备间苯三酚的体外合成体系。该体系的性能表明，体外合成间苯三酚具有一定的优势，有可能成为一种可行的工业替代品。综上所述，体外生物系统将为醋酸酯生产重要工业化学品提供一个可行的选择，它可以作为一个多功能的生物合成平台。
BackgroundPhloroglucinol is an important chemical, and the biosynthesis processes which can convert glucose to phloroglucinol have been established. However, due to approximate 80% of the glucose being transformed into undesirable by-products and biomass, this biosynthesis process only shows a low yield with the highest value of about 0.20 g/g. The industrial applications are usually hindered by the low current productivity and yield and also by the high costs. Generally, several different aspects limit the development of phloroglucinol biosynthesis. The yield of phloroglucinol is one of the most important parameters for its bioconversion especially from economic and ecological points of view. The in vitro biosynthesis of bio-based chemicals, is a flexible alternative with potentially high-yield to in vivo biosynthetic technology.ResultsBy comparing the activity of acetyl-CoA synthetase (ACS) fromEscherichia coliandAcetobacter pasteurianus, the highly active ACS2 was identified inA. pasteurianus. Acetyl-CoA carboxylase (ACC) fromAcinetobacter calcoaceticusand phloroglucinol synthase (PhlD) fromPseudomonas fluorescenspf-5 were expressed and purified. Acetate was successfully transformed into phloroglucinol by the combined activity of above-mentioned enzymes and required cofactor. After optimization of the in vitro reaction system, phloroglucinol was then produced with a yield of nearly 0.64 g phloroglucinol/g acetic acid, which was equal to 91.43% of the theoretically possible maximum.ConclusionsIn this work, a novel in vitro synthetic system for a highly efficient production of phloroglucinol from acetate was demonstrated. The system’s performance suggests that in vitro synthesis of phloroglucinol has some advantages and is potential to become a feasible industrial alternative. Based on the results presented herewith, it is believed that in vitro biosystem will provide a feasible option for production of important industrial chemicals from acetate, which could work as a versatile biosynthetic platform.
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影响因子：
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作者：
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通讯作者：
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