Production of D-lactic acid containing polyhydroxyalkanoate polymers in yeast Saccharomyces cerevisiae

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Volume Title

A1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä

Date

2021-06

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en

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12

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Journal of Industrial Microbiology and Biotechnology, Volume 48, issue 5-6

Abstract

Polyhydroxyalkanoates (PHAs) provide biodegradable and bio-based alternatives to conventional plastics. Incorporation of 2-hydroxy acid monomers into polymer, in addition to 3-hydroxy acids, offers possibility to tailor the polymer properties. In this study, poly(D-lactic acid) (PDLA) and copolymer P(LA-3HB) were produced and characterized for the first time in the yeast Saccharomyces cerevisiae. Expression of engineered PHA synthase PhaC1437(Ps6-19), propionyl-CoA transferase Pct540Cp, acetyl-CoA acetyl-transferase PhaA, and acetoacetyl-CoA reductase PhaB1 resulted in accumulation of 3.6% P(LA-3HB) and expression of engineered enzymes PhaC1Pre and PctMe resulted in accumulation of 0.73% PDLA of the cell dry weight (CDW). According to NMR, P(LA-3HB) contained D-lactic acid repeating sequences. For reference, expression of PhaA, PhaB1, and PHA synthase PhaC1 resulted in accumulation 11% poly(hydroxybutyrate) (PHB) of the CDW. Weight average molecular weights of these polymers were comparable to similar polymers produced by bacterial strains, 24.6, 6.3, and 1 130 kDa for P(LA-3HB), PDLA, and PHB, respectively. The results suggest that yeast, as a robust and acid tolerant industrial production organism, could be suitable for production of 2-hydroxy acid containing PHAs from sugars or from 2-hydroxy acid containing raw materials. Moreover, the wide substrate specificity of PHA synthase enzymes employed increases the possibilities for modifying copolymer properties in yeast in the future.

Description

This work was supported by the Maj Ja Tor Nessling säätiö and Tor Nessling Foundation (grant numbers 201700259, 201800005) to A.Y., Academy of Finland (project 310191) to M.T., H.M., A.A.-H.; VTT Technical Research Centre of Finland, and Centre for Young Synbio Scientists (CYSS) funded by Jenny ja Antti Wihurin rahasto and Antti Wihuri Foundation to A.Y.

Keywords

Poly(D-lactic acid), PHB, Copolymer, Biopolymer, Saccharomyces cerevisiae, PROPIONATE COA-TRANSFERASE, POLYLACTIC ACID, LACTATE, BIOSYNTHESIS

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Citation

Ylinen, A, Maaheimo, H, Anghelescu-Hakala, A, Penttilä, M, Salusjarvi, L & Toivari, M 2021, ' Production of D-lactic acid containing polyhydroxyalkanoate polymers in yeast Saccharomyces cerevisiae ', Journal of Industrial Microbiology and Biotechnology, vol. 48, no. 5-6, kuab028 . https://doi.org/10.1093/jimb/kuab028