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Production of C4 and C5 branched-chain alcohols by engineered Escherichia. coli

  • Bioenergy/Biofuels/Biochemicals
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Higher alcohols, longer chain alcohols, contain more than 3 carbon atoms, showed close energy advantages as gasoline, and were considered as the next generation substitution for chemical fuels. Higher alcohol biosynthesis by native microorganisms mainly needs gene expression of heterologous keto acid decarboxylase and alcohol dehydrogenases. In the present study, branched-chain α-keto acid decarboxylase gene from Lactococcus lactis subsp. lactis CICC 6246 (Kivd) and alcohol dehydrogenases gene from Zymomonas mobilis CICC 41465 (AdhB) were transformed into Escherichia coli for higher alcohol production. SDS-PAGE results showed these two proteins were expressed in the recombinant strains. The resulting strain was incubated in LB medium at 37 °C in Erlenmeyer flasks and much more 3-methyl-1-butanol (104 mg/L) than isobutanol (24 mg/L) was produced. However, in 5 g/L glucose-containing medium, the production of two alcohols was similar, 156 and 161 mg/L for C4 (isobutanol) and C5 (3-methyl-1-butanol) alcohol, respectively. Effects of fermentation factors including temperature, glucose content, and α-keto acid on alcohol production were also investigated. The increase of glucose content and the adding of α-keto acids facilitated the production of C4 and C5 alcohols. The enzyme activities of pure Kivd on α-ketoisovalerate and α-ketoisocaproate were 26.77 and 21.24 μmol min−1 mg−1, respectively. Due to its ability on decarboxylation of α-ketoisovalerate and α-ketoisocaproate, the recombinant E. coli strain showed potential application on isoamyl alcohol and isobutanol production.

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Acknowledgments

This research was financially supported by National Natural Science Foundation of China (No. 2117623 and No. 21211140237), National High Technology Research and Development Program of China (863 Program) (2013AA065803), Guangdong science and technology research program (2013B010403021), Guangzhou science and technology research program (2013J4300026) and Foundation for Innovative Research of Guangzhou Institute of Energy Conversion, CAS (y407p11001).

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    Authors and Affiliations

    1. Key Laboratory of Renewable Energy and Gas Hydrate, Chinese Academy of Sciences, Guangzhou, 510640, China

      Xiaoyan Chen, Jingliang Xu, Liu Yang, Zhenhong Yuan, Shiyuan Xiao, Yu Zhang, Cuiyi Liang, Minchao He & Ying Guo

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    1. Xiaoyan Chen
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    Correspondence to Zhenhong Yuan.

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    X. Chen and J. Xu contribute equally to this work.

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    Chen, X., Xu, J., Yang, L. et al. Production of C4 and C5 branched-chain alcohols by engineered Escherichia. coli . J Ind Microbiol Biotechnol 42, 1473–1479 (2015). https://doi.org/10.1007/s10295-015-1656-z

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    • DOI: https://doi.org/10.1007/s10295-015-1656-z

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