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. 2015 Oct;119(4):1057-63.
doi: 10.1111/jam.12905. Epub 2015 Aug 21.

Production of adipic acid by the native-occurring pathway in Thermobifida fusca B6

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Production of adipic acid by the native-occurring pathway in Thermobifida fusca B6

Y Deng et al. J Appl Microbiol. 2015 Oct.

Abstract

Aims: To identify the native-occurring adipate pathway in a previously engineered Thermobifida fusca B6 strain and optimize the adipic acid production of this strain on glucose and corncob.

Methods and results: The native-occurring adipate pathway in T. fusca B6 was identified to be the reverse adipate degradation pathway, including five steps: β-ketothiolase (Tfu_0875), 3-hydroxyacyl-CoA dehydrogenase (Tfu_2399), 3-hydroxyadipyl-CoA dehydrogenase (Tfu_0067), 5-Carboxy-2-pentenoyl-CoA reductase (Tfu_1647) and succinyl-CoA synthetase (Tfu_2576, Tfu_2577). The cell lysates of T. fusca wild-type strain with the addition of the Tfu_1647 protein produced trace adipic acid, while no adipic acid was produced in the absence of this protein. The above results prove that the low expression of Tfu_1647 in the wild-type strain was the reason why it did not produce any adipic acid. We then demonstrated that in T. fusca B6, the maximal titre of adipic acid on 50 g l(-1) glucose was 2·23 g l(-1) with 0·045 g g(-1) -glucose yield and 0·22 g l(-1) adipic acid was produced from 19·38 g l(-1) milled corncob.

Conclusions: The reverse adipate degradation pathway was found to be responsible for the adipate synthesis in T. fusca B6 and Tfu_1647 was the regulatory node on this pathway.

Significance and impact of the study: Thermobifida fusca B6 was the first reported micro-organism using its native-occurring pathway to accumulate adipic acid. It had the highest reported yield and titre of adipic acid so far.

Keywords: 5-Carboxy-2-pentenoyl-CoA reductase; T. fusca B6; adipic acid; corncob; reverse adipate degradation pathway.

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