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

  • Xiaoxia Zhang
  • Yufeng Mao
  • Baowei Wang
  • Zhenzhen Cui
  • Zhidan Zhang
  • Zhiwen Wang
  • Tao ChenEmail author
Biocatalysis - Original Paper
  • 75 Downloads

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

Lactoyl-CoA CoA-transferases Kinetic characterization Megasphaera elsdenii 

Notes

Acknowledgements

This research was financially supported by the National Natural Science Foundation of China (NSFC-21776208, NSFC-21621004 and NSFC-21776209).

Compliance with ethical standards

Conflict of interest

The authors declare no conflicts of interest.

Supplementary material

10295_2019_2174_MOESM1_ESM.docx (2.9 mb)
Supplementary material 1 (DOCX 3015 kb)

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Copyright information

© Society for Industrial Microbiology and Biotechnology 2019

Authors and Affiliations

  1. 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 TechnologyTianjin UniversityTianjinPeople’s Republic of China
  2. 2.Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial BiotechnologyChinese Academy of SciencesTianjinPeople’s Republic of China

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