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Biochemical properties and physiological roles of NADP-dependent malic enzyme in Escherichia coli

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Abstract

Malic enzymes catalyze the reversible oxidative decarboxylation of L-malate using NAD(P)+ as a cofactor. NADP-dependent malic enzyme (MaeB) from Escherichia coli MG1655 was expressed and purified as a fusion protein. The molecular weight of MaeB was about 83 kDa, as determined by SDS-PAGE. The recombinant MaeB showed a maximum activity at pH 7.8 and 46°C. MaeB activity was dependent on the presence of Mn2+ but was strongly inhibited by Zn2+. In order to understand the physiological roles, recombinant E. coli strains (icd NADP/ΔmaeB and icd NAD/ΔmaeB) containing NADP-dependent isocitrate dehydrogenase (IDH), or engineered NAD-dependent IDH with the deletion of the maeB gene, were constructed using homologous recombination. During growth on acetate, icd NAD/ΔmaeB grew poorly, having a growth rate only 60% that of the wild-type strain (icd NADP). Furthermore, icd NADP/ΔmaeB exhibited a 2-fold greater adaptability to acetate than icd NAD/ΔmaeB, which may be explained by more NADPH production for biosynthesis in icd NADP/ΔmaeB due to its NADP-dependent IDH. These results indicated that MaeB was important for NADPH production for bacterial growth on acetate. We also observed that MaeB activity was significantly enhanced (7.83-fold) in icd NAD, which was about 3-fold higher than that in icd NADP, when switching from glucose to acetate. The marked increase of MaeB activity was probably induced by the shortage of NADPH in icd NAD. Evidently, MaeB contributed to the NADPH generation needed for bacterial growth on two carbon compounds.

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Authors and Affiliations

  1. Key Laboratory of Molecular Evolution and Biodiversity and Institute of Molecular Biology and Biotechnology, Anhui Normal University, Wuhu, 241000, P. R. China

    Baojuan Wang, Peng Wang, Enxia Zheng, Xiangxian Chen, Ping Song, Ruirui Su & Guoping Zhu

  2. The Clinical Laboratory of Wuhu Second People’s Hospital, Wuhu, 241000, P. R. China

    Hanjun Zhao

  3. Yijishan Hospital of Wannan Medical College, Wuhu, 241000, P. R. China

    Xiaoning Li

Authors
  1. Baojuan Wang
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  2. Peng Wang
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  3. Enxia Zheng
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  4. Xiangxian Chen
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  5. Hanjun Zhao
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  6. Ping Song
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  7. Ruirui Su
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  8. Xiaoning Li
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  9. Guoping Zhu
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Correspondence to Guoping Zhu.

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These authors contributed equally to this work.

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Wang, B., Wang, P., Zheng, E. et al. Biochemical properties and physiological roles of NADP-dependent malic enzyme in Escherichia coli . J Microbiol. 49, 797–802 (2011). https://doi.org/10.1007/s12275-011-0487-5

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  • DOI: https://doi.org/10.1007/s12275-011-0487-5

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