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Enhanced butanol production in Clostridium acetobutylicum ATCC 824 by double overexpression of 6-phosphofructokinase and pyruvate kinase genes

  • Bioenergy and biofuels
  • Published:
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Abstract

This study elucidated the importance of two critical enzymes in the regulation of butanol production in Clostridium acetobutylicum ATCC 824. Overexpression of both the 6-phosphofructokinase (pfkA) and pyruvate kinase (pykA) genes increased intracellular concentrations of ATP and NADH and also resistance to butanol toxicity. Marked increases of butanol and ethanol production, but not acetone, were also observed in batch fermentation. The butanol and ethanol concentrations were 29.4 and 85.5 % higher, respectively, in the fermentation by double-overexpressed C. acetobutylicum ATCC 824/pfkA+pykA than the wild-type strain. Furthermore, when fed-batch fermentation using glucose was carried out, the butanol and total solvent (acetone, butanol, and ethanol) concentrations reached as high as 19.12 and 28.02 g/L, respectively. The reason for improved butanol formation was attributed to the enhanced NADH and ATP concentrations and increased tolerance to butanol in the double-overexpressed strain.

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Abbreviations

hsdR :

Host-specific restriction deficient

mcrA :

Methylcytosine-specific restriction deleted

recA1 :

Homologous recombination deleted

endA1 :

Endonuclease deleted

p15A ori :

p15 origin of replication

Cmr :

Chloramphenicol resistance

Φ3TI:

B. subtilis phage Φ3TI methyltransferase gene

ColE1:

ColE1 origin of replication

pIM13:

pIM13 gram-positive origin of replication

Apr :

Ampicillin resistance

MLSr :

Macrolide-lincosamide-streptogramin B resistance

P-ptb :

Clostridial phosphate butyryltransferase promoter

ctfA/B :

Acetoacetyl-CoA transferase genes

adc :

AADC gene

pfkA :

6-phosphofuctokinase gene

pykA :

Pyruvate kinase gene

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Acknowledgment

The authors acknowledge the financial support provided by the Korean Ministry of Environment (405-112-039).

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Authors and Affiliations

  1. Department of Environmental Engineering and Energy, Myongji University, 116 Myongji-Ro, Cheoin-Gu, Yongin, Gyeonggi-Do, 449-728, Republic of Korea

    Jey-R S. Ventura & Deokjin Jahng

  2. Shanghai Acebright Phamaceuticals Co. Ltd., Department 9, No. 2 Zhangheng Road, Pudong New Area, Shanghai, 201203, China

    Hui Hu

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  1. Jey-R S. Ventura
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  2. Hui Hu
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Correspondence to Deokjin Jahng.

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Ventura, JR.S., Hu, H. & Jahng, D. Enhanced butanol production in Clostridium acetobutylicum ATCC 824 by double overexpression of 6-phosphofructokinase and pyruvate kinase genes. Appl Microbiol Biotechnol 97, 7505–7516 (2013). https://doi.org/10.1007/s00253-013-5075-7

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  • DOI: https://doi.org/10.1007/s00253-013-5075-7

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