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Proteome-based physiological analysis of the metabolically engineered succinic acid producer Mannheimia succiniciproducens LPK7

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Abstract

Using two-dimensional gel electrophoresis (2-DE) and mass spectrometry, the proteome of a metabolically engineered succinic acid-overproducing bacterium, Mannheimia succiniciproducens LPK7, was examined and compared with that of its wild type strain, MBEL55E, to elucidate the physiological and metabolic changes responsible for succinic acid overproduction and cell growth. Comparative proteomic studies clearly showed that the expression levels of enzymes involved in the ATP formation and consumption (AtpD, Ppa, SerS, ProS, Pnp, PotD, MalK, RbsB, and TbpA), pyruvate metabolism (AceF and Lpd), glycolysis (GapA, Pgk, Fba, and TpiA), and amino acid biosynthesis (Asd, DapA, DapD, Gdh, ArgD, and ArgG) varied significantly in the LPK7 strain compared with those in the MBEL55E strain. Based on the comparative proteome profiling, the formation of pyruvic acid, a newly formed byproduct in the engineered LPK7 strain, could be reduced by adding into the culture medium pantothenate and l-cysteine, which serve as precursors of CoA biosynthesis.

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Acknowledgments

The authors thank Hyohak Song at KAIST and Jong Shin Yoo at Korea Basic Science Institute (KBSI) for their helps on manuscript writing and protein identification, respectively. This work was supported by the Genome-based Integrated Bioprocess Development Project of the Ministry of Education, Science and Technology. Further supports by the LG Chem Chair Professorship, Brain Korea 21 project, and by the KOSEF through the Center for Ultramicrochemical Process Systems are appreciated.

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Authors and Affiliations

  1. Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering (BK21 Program), BioProcess Engineering Research Center, KAIST, 335 Gwahangno, Yuseong-gu, Daejeon, 305-701, Republic of Korea

    Jeong Wook Lee & Sang Yup Lee

  2. Center for Systems and Synthetic Biotechnology, Institute for the BioCentury, KAIST, 335 Gwahangno, Yuseong-gu, Daejeon, 305-701, Republic of Korea

    Jeong Wook Lee & Sang Yup Lee

  3. Department of Bio and Brain Engineering and Bioinformatics Research Center, KAIST, 335 Gwahangno, Yuseong-gu, Daejeon, 305-701, Republic of Korea

    Sang Yup Lee

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  1. Jeong Wook Lee
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Correspondence to Sang Yup Lee.

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Lee, J.W., Lee, S.Y. Proteome-based physiological analysis of the metabolically engineered succinic acid producer Mannheimia succiniciproducens LPK7. Bioprocess Biosyst Eng 33, 97–107 (2010). https://doi.org/10.1007/s00449-009-0339-4

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