Engineering the Yeast Yarrowia lipolytica for Production of Polylactic Acid Homopolymer

Frontiers in Bioengineering and Biotechnology
Sophie LajusFlorence Bordes

Abstract

Polylactic acid is a plastic polymer widely used in different applications from printing filaments for 3D printer to mulching films in agriculture, packaging materials, etc. Here, we report the production of poly-D-lactic acid (PDLA) in an engineered yeast strain of Yarrowia lipolytica. Firstly, the pathway for lactic acid consumption in this yeast was identified and interrupted. Then, the heterologous pathway for PDLA production, which contains a propionyl-CoA transferase (PCT) converting lactic acid into lactyl-CoA, and an evolved polyhydroxyalkanoic acid (PHA) synthase polymerizing lactyl-CoA, was introduced into the engineered strain. Among the different PCT proteins that were expressed in Y. lipolytica, the Clostridium propionicum PCT exhibited the highest efficiency in conversion of D-lactic acid to D-lactyl-CoA. We further evaluated the lactyl-CoA and PDLA productions by expressing this PCT and a variant of Pseudomonas aeruginosa PHA synthase at different subcellular localizations. The best PDLA production was obtained by expressing the PCT in the cytosol and the variant of PHA synthase in peroxisome. PDLA homopolymer accumulation in the cell reached 26 mg/g-DCW, and the molecular weights of the polymer (Mw = 50.5 × 103 ...Continue Reading

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Citations

Dec 4, 2020·Proceedings of the National Academy of Sciences of the United States of America·Simon DusséauxSotirios C Kampranis
Apr 27, 2021·Journal of Industrial Microbiology & Biotechnology·Anna YlinenMervi Toivari
Aug 5, 2021·Applied Biochemistry and Biotechnology·Shivam BholaPradeep Kumar
Aug 29, 2021·FEMS Yeast Research·Young-Kyoung ParkPatrick Fickers
Oct 26, 2021·Bioresource Technology·Hideo KawaguchiAkihiko Kondo

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Datasets Mentioned

BETA
AFJ29290.1
EAA58342.1
CAJ93797.1
CAB77207.1

Methods Mentioned

BETA
PCR
NMR
PCT
fluorescence microscopy
nuclear magnetic resonance

Software Mentioned

Interproscan
Trace Finder
Clustal Omega
Tracefinder
Mitoprot
Pfam
GenScript
IsoCor

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