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Succinate Production with Metabolically Engineered Escherichia coli Using Elephant Grass Stalk (Pennisetum purpureum) Hydrolysate as Carbon Source

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Abstract

Succinic acid is a spectacular chemical that can be used as the precursor of various industrial products including pharmaceuticals and biochemicals. The improvement of the succinic acid market depends on strains engineering that is capable of producing succinic acid at high yield and excellent growth rate which could utilize the wide range of carbon sources such as renewable biomass. Here we use counter selection using catAsacB for pathway design and strains developments. In this investigation, metabolically engineered Escherichia coli M6PM strain was constructed for the synthesis of succinic acid using elephant grass stalk (Pennisetum purpureum) as a carbon source. Elephant grass stalk hydrolysate was prepared which comprised of 11.60 ± 0.04 g/L glucose, 27.22 ± 0.04 g/L xylose and 0.65 ± 0.04 g/L arabinose. Metabolically engineered E. coli M6PM was constructed and fermentation with pure sugars revealed that it could utilize xylose and glucose efficiently. E. coli M6PM produced a final succinate concentration of 30.03 ± 0.02 g/L and a yield of 1.09 mol/mol during 72 h dual-phase fermentation using elephant grass stalk hydrolysate, which resulted in 64% maximum theoretical yield of succinic acid. The high succinate yield from elephant grass stalk demonstrated possible application of renewable biomass as feedstock for the synthesis of succinic acid using recombinant E. coli.

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Abbreviations

HPLC:

High performance liquid chromatography

O.D:

Optical density

rpm:

Revolution per minutes

IPTG:

l isopropyl-β-d-thiogalactopyranoside

ldhA:

Lactate dehydrogenase A

pta-ackA:

Phosphotranacetylase acetate kinase A

pflB:

Pyruvate formate lyase B

poxB:

Pyruvate oxidase B

pgi :

Phosphoglucose isomerase

mreC:

Murein cluster C

pyc :

Pyruvate carboxylase

ppc :

Phosphoenol pyruvate carboxylase

zwf :

Glucose 6-phosphate dehydrogenase

pgl :

6-Phosphogluconolactonase

gnd :

6-Phosphogluconate dehydrogenase

tkt :

Transketolase

tal :

Transaldolase

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Acknowledgements

This work was supported by National High Technology Research and Development Program of China (863 Project, No 2014AA021905).

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Authors and Affiliations

  1. Key Laboratory of Green Process and Engineering, State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, 100190, People’s Republic of China

    Ayobami Matthew Olajuyin, Maohua Yang, Tingzhen Mu, Moustafa Mohamed Sharshar & Jianmin Xing

  2. University of Chinese Academy of Sciences, Beijing, 100049, People’s Republic of China

    Ayobami Matthew Olajuyin, Maohua Yang, Tingzhen Mu, Moustafa Mohamed Sharshar & Jianmin Xing

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  1. Ayobami Matthew Olajuyin
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  2. Maohua Yang
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Correspondence to Jianmin Xing.

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Olajuyin, A.M., Yang, M., Mu, T. et al. Succinate Production with Metabolically Engineered Escherichia coli Using Elephant Grass Stalk (Pennisetum purpureum) Hydrolysate as Carbon Source. Waste Biomass Valor 11, 1717–1725 (2020). https://doi.org/10.1007/s12649-018-0524-2

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  • DOI: https://doi.org/10.1007/s12649-018-0524-2

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