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Elimination of carbon catabolite repression in Klebsiella oxytoca for efficient 2,3-butanediol production from glucose–xylose mixtures

  • Applied Genetics and Molecular Biotechnology
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Abstract

Microbial preference for glucose implies incomplete and/or slow utilization of lignocellulose hydrolysates, which is caused by the regulatory mechanism named carbon catabolite repression (CCR). In this study, a 2,3-butanediol (2,3-BD) producing Klebsiella oxytoca strain was engineered to eliminate glucose repression of xylose utilization. The crp(in) gene, encoding the mutant cyclic adenosine monophosphate (cAMP) receptor protein CRP(in), which does not require cAMP for functioning, was characterized and overexpressed in K. oxytoca. The engineered recombinant could utilize a mixture of glucose and xylose simultaneously, without CCR. The profiles of sugar consumption and 2,3-BD production by the engineered recombinant, in glucose and xylose mixtures, were examined and showed that glucose and xylose could be consumed simultaneously to produce 2,3-BD. This study offers a metabolic engineering strategy to achieve highly efficient utilization of sugar mixtures derived from the lignocellulosic biomass for the production of bio-based chemicals using enteric bacteria.

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Acknowledgements

We are grateful to Prof. Mike J. Merrick, John Innes Centre, UK, and Prof. Lonnie O. Ingram, University of Florida, USA, for providing the pDK7 and pLOI 3244 plasmids. This work was financially supported by the National Natural Science Foundation of China (Nos. 20606018 and 21006049), the Key Program of National Natural Science Foundation of China (No. 20936002), the National Basic Research Program of China (Nos. 2007CB707805, 2009CB724700 and 2011CB200906), and the Program for New Century Excellent Talents in University from the Ministry of Education of China (No. NCET-09-0157). X.-J. Ji was supported by the Innovation Fund for Doctoral Dissertation of Nanjing University of Technology (No. BSCX200808), the China Postdoctoral Science Foundation Funded Project (No. 20100471328) and the Jiangsu Planned Projects for Postdoctoral Research Funds of China (No. 1001015 C).

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Authors and Affiliations

  1. State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, No. 5 Xinmofan Road, Nanjing, 210009, People’s Republic of China

    Xiao-Jun Ji, Zhi-Kui Nie, He Huang, Lu-Jing Ren, Chao Peng & Ping-Kai Ouyang

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  1. Xiao-Jun Ji
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  2. Zhi-Kui Nie
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Correspondence to He Huang.

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Supplementary Fig. S1

Alignments of cloned nucleotide sequence containing coding regions for crp(in) from Escherichia coli ET25 (GenBank accession no. HM595439) and crp from E. coli K12 (NC000913), Klebsiella pneumoniae MGH 78578 (CP000647), Klebsiella oxytoca M5al (AJ278967), and Klebsiella aerogenes KC1043 (M68973) searched from GenBank data. Nucleotides differing in E. coli ET25 are surrounded with rectangles (DOC 1,259 kb)

Supplementary Fig. S2

Alignments of amino acid sequences of CRP(in) of Escherichia coli ET25 (ADK89557.1) and CRP of E. coli K12 (AP_004432), Klebsiella pneumoniae MGH 78578 (YP 001337397), Klebsiella oxytoca M5al (CAC07215) and Klebsiella aerogenes KC1043 (AAA25058). Amino acids differing in E. coli ET25 are surrounded with rectangles (DOC 1,140 kb)

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Ji, XJ., Nie, ZK., Huang, H. et al. Elimination of carbon catabolite repression in Klebsiella oxytoca for efficient 2,3-butanediol production from glucose–xylose mixtures. Appl Microbiol Biotechnol 89, 1119–1125 (2011). https://doi.org/10.1007/s00253-010-2940-5

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