Molecular Biotechnology

, Volume 45, Issue 2, pp 121–128 | Cite as

Molecular Cloning and Characterization of a Malic Enzyme Gene from the Oleaginous Yeast Lipomyces starkeyi

  • Wei Tang
  • Sufang Zhang
  • Haidong Tan
  • Zongbao K. Zhao


The malic enzyme-encoding cDNA (GQ372891) from the oleaginous yeast Lipomyces starkeyi AS 2.1560 was isolated, which has an 1719-bp open reading frame flanked by a 290-bp 5′ untranslated sequence and a 92-bp 3′ untranslated sequence. The proposed gene, LsME1, encoded a protein with 572 amino acid residues. The protein presented 58% sequence identity with the malic enzymes from Yarrowia lipolytica CLIB122 and Aspergillus fumigatus Af293. The LsME1 gene was cloned into the vector pMAL-p4x to express a fusion protein (MBP-LsME1) in Escherichia coli TB1. The fusion protein was purified and then cleaved by Factor Xa to give the recombinant LsME1. This purified enzyme took either NAD+ or NADP+ as the coenzyme but preferred NAD+. The Km values for malic acid, NAD+ and NADP+ were 0.85 ± 0.05 mM, 0.34 ± 0.08 mM, and 7.4 ± 0.32 mM, respectively, at pH 7.3.


Lipomyces starkeyi Malic enzyme RACE Protein expression Enzyme kinetics 


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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Wei Tang
    • 1
    • 2
  • Sufang Zhang
    • 1
  • Haidong Tan
    • 1
  • Zongbao K. Zhao
    • 1
  1. 1.Dalian Institute of Chemical Physics, CASDalianPeople’s Republic of China
  2. 2.Graduate School of the Chinese Academy of SciencesBeijingPeople’s Republic of China

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