Exploring functionality of the reverse β-oxidation pathway in Corynebacterium glutamicum for production of adipic acid
Journal article, 2021

Background: Adipic acid, a six-carbon platform chemical mainly used in nylon production, can be produced via reverse β-oxidation in microbial systems. The advantages posed by Corynebacterium glutamicum as a model cell factory for implementing the pathway include: (1) availability of genetic tools, (2) excretion of succinate and acetate when the TCA cycle becomes overflown, (3) initiation of biosynthesis with succinyl-CoA and acetyl-CoA, and (4) established succinic acid production. Here, we implemented the reverse β-oxidation pathway in C. glutamicum and assessed its functionality for adipic acid biosynthesis. Results: To obtain a non-decarboxylative condensation product of acetyl-CoA and succinyl-CoA, and to subsequently remove CoA from the condensation product, we introduced heterologous 3-oxoadipyl-CoA thiolase and acyl-CoA thioesterase into C. glutamicum. No 3-oxoadipic acid could be detected in the cultivation broth, possibly due to its endogenous catabolism. To successfully biosynthesize and secrete 3-hydroxyadipic acid, 3-hydroxyadipyl-CoA dehydrogenase was introduced. Addition of 2,3-dehydroadipyl-CoA hydratase led to biosynthesis and excretion of trans-2-hexenedioic acid. Finally, trans-2-enoyl-CoA reductase was inserted to yield 37 µg/L of adipic acid. Conclusions: In the present study, we engineered the reverse β-oxidation pathway in C. glutamicum and assessed its potential for producing adipic acid from glucose as starting material. The presence of adipic acid, albeit small amount, in the cultivation broth indicated that the synthetic genes were expressed and functional. Moreover, 2,3-dehydroadipyl-CoA hydratase and β-ketoadipyl-CoA thiolase were determined as potential target for further improvement of the pathway.

Author

Jae Ho Shin

Chalmers, Biology and Biological Engineering, Industrial Biotechnology

Aaron John Christian Andersen

Technical University of Denmark (DTU)

Puck Achterberg

Student at Chalmers

Delft University of Technology

Lisbeth Olsson

Chalmers, Biology and Biological Engineering, Industrial Biotechnology

Microbial Cell Factories

1475-2859 (ISSN)

Vol. 20 1 155

A novel strategy for cell factory design applied to adipic acid production - combining synthetic pathway and electrofermentation

Swedish Research Council (VR) (2016-03344), 2017-01-01 -- 2021-12-31.

Subject Categories

Microbiology

Organic Chemistry

Other Industrial Biotechnology

DOI

10.1186/s12934-021-01647-7

PubMed

34348702

More information

Latest update

8/19/2021