2. Academic Validation
  3. Screening, expression, purification and characterization of CoA-transferases for lactoyl-CoA generation

Screening, expression, purification and characterization of CoA-transferases for lactoyl-CoA generation

  • J Ind Microbiol Biotechnol. 2019 Jul;46(7):899-909. doi: 10.1007/s10295-019-02174-6.
Xiaoxia Zhang 1,  Yufeng Mao 2,  Baowei Wang 1,  Zhenzhen Cui 1,  Zhidan Zhang 2,  Zhiwen Wang 1,  Tao Chen 3
Affiliations

Affiliations

  • 1 Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, People's Republic of China.
  • 2 Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, People's Republic of China.
  • 3 Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, People's Republic of China. [email protected].
PMID: 30963328 DOI: 10.1007/s10295-019-02174-6
Abstract

Lactoyl-CoA is critical for the biosynthesis of biodegradable and biocompatible lactate-based copolymers, which have wide applications. However, reports on acetyl-CoA: lactate CoA-transferases (ALCTs) are rare. To exploit novel ALCTs, amino acid sequence similarity searches based on the CoA-transferases from Clostridium propionicum and Megasphaera elsdenii were conducted. Two known and three novel enzymes were expressed, purified and characterized. Three novel ALCTs were identified, one each from Megasphaera sp. DISK 18, Clostridium lactatifermentans An75 and Firmicutes bacterium CAG: 466. ME-PCT from Megasphaera elsdenii had the highest catalytic efficiency for both acetyl-CoA (264.22 s-1 mM-1) and D-lactate (84.18 s-1 mM-1) with a broad temperature range for activity and good stability. This study, therefore, offers novel and efficient enzymes for lactoyl-CoA generation. To our best knowledge, this is the first report on the systematic mining of ALCTs, which offers valuable new tools for the engineering of pathways that rely on these enzymes.

Keywords

CoA-transferases; Kinetic characterization; Lactoyl-CoA; Megasphaera elsdenii.

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