A bio-based route to the carbon-5 chemical glutaric acid and to bionylon-6,5 using metabolically engineered Corynebacterium glutamicum,Green Chemistry

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A bio-based route to the carbon-5 chemical glutaric acid and to bionylon-6,5 using metabolically engineered Corynebacterium glutamicum
Green Chemistry ( IF 9.3 ) Pub Date : 2018-09-24 , DOI: 10.1039/c8gc01901k
Christina Maria Rohles _{1,

2,

3,

4} , Lars Gläser _{1,

2,

3,

4} , Michael Kohlstedt _{1,

2,

3,

4} , Gideon Gießelmann _{1,

2,

3,

4} , Samuel Pearson _{3,

4,

5} , Aránzazu del Campo _{3,

4,

5} , Judith Becker _{1,

2,

3,

4} , Christoph Wittmann _{1,

2,

3,

4}

Affiliation

In the present work, we established the bio-based production of glutarate, a carbon-5 dicarboxylic acid with recognized value for commercial plastics and other applications, using metabolically engineered Corynebacterium glutamicum. The mutant C. glutamicum AVA-2 served as a starting point for strain development, because it secreted small amounts of glutarate as a consequence of its engineered 5-aminovalerate pathway. Starting from AVA-2, we overexpressed 5-aminovalerate transaminase (gabT) and glutarate semialdehyde dehydrogenase (gabD) under the control of the constitutive tuf promoter to convert 5-aminovalerate further to glutarate. The created strain GTA-1 formed glutarate as a major product, but still secreted 5-aminovalerate as well. This bottleneck was tackled at the level of 5-aminovalerate re-import. The advanced strain GTA-4 overexpressed the newly discovered 5-aminovalerate importer NCgl0464 and formed glutarate from glucose in a yield of 0.27 mol mol⁻¹. In a fed-batch process, GTA-4 produced more than 90 g L⁻¹ glutarate from glucose and molasses based sugars in a yield of up to 0.70 mol mol⁻¹ and a maximum productivity of 1.8 g L⁻¹ h⁻¹, while 5-aminovalerate was no longer secreted. The bio-based glutaric acid was purified to >99.9% purity. Interfacial polymerization and melt polymerization with hexamethylenediamine yielded bionylon-6,5, a polyamide with a unique structure.

中文翻译：

使用代谢工程化的谷氨酸棒杆菌，生物合成碳-5化学戊二酸和生物尼龙-6,5的途径

在目前的工作中，我们使用谷氨酸棒状杆菌进行了生物工程生产谷氨酸，这是一种在商业塑料和其他应用中具有公认价值的戊二酸碳五碳。突变体谷氨酸棒杆菌AVA-2用作菌株开发的起点，因为其工程化的5-氨基戊酸途径可分泌少量谷氨酸。从AVA-2开始，我们在组成性tuf的控制下过表达5-氨基戊酸转氨酶（gabT）和谷氨酸半醛脱氢酶（gabD）。启动子将5-氨基戊酸酯进一步转化为谷氨酸。产生的菌株GTA-1形成谷氨酸作为主要产物，但仍分泌5-氨基戊酸酯。解决了5-氨基戊酸酯再进口水平的这一瓶颈。先进菌株GTA-4过表达新发现的5-氨基戊酸酯输入物NCgl0464，并由葡萄糖形成戊二酸酯，收率为0.27mol mol ^-1。在分批补料工艺中，GTA-4从葡萄糖和糖蜜基糖中制得了超过90 g L ^-1的戊二酸酯，产率高达0.70 mol mol ^-1，最大生产率为1.8 g L ^-1 h ^-1，而不再分泌5-氨基戊酸。将生物基戊二酸纯化至> 99.9％的纯度。用六亚甲基二胺进行界面聚合和熔融聚合，得到生物尼龙-6,5，这是一种具有独特结构的聚酰胺。

更新日期：2018-10-15

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