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

Anna Ylinen; Hannu Maaheimo; Adina Anghelescu-Hakala; Merja Penttilä; Laura Salusjärvi; Mervi Toivari

doi:10.1093/jimb/kuab028

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

^*Corresponding author for this work

Aalto University

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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 PhaC1437Ps6-19, propionyl-CoA transferase Pct540Cp, acetyl-CoA acetyltransferase 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.

Original language	English
Article number	kuab028
Journal	Journal of industrial microbiology and biotechnology
Volume	48
Issue number	5-6
DOIs	https://doi.org/10.1093/jimb/kuab028
Publication status	Published - 1 Jun 2021
MoE publication type	A1 Journal article-refereed

Funding

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

Biopolymer
Copolymer
PHB
Poly(D-lactic acid)
Saccharomyces cerevisiae

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10.1093/jimb/kuab028Licence: CC BY

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title = "Production of D-lactic acid containing polyhydroxyalkanoate polymers in yeast Saccharomyces cerevisiae",

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 PhaC1437Ps6-19, propionyl-CoA transferase Pct540Cp, acetyl-CoA acetyltransferase 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.",

keywords = "Biopolymer, Copolymer, PHB, Poly(D-lactic acid), Saccharomyces cerevisiae",

author = "Anna Ylinen and Hannu Maaheimo and Adina Anghelescu-Hakala and Merja Penttil{\"a} and Laura Salusj{\"a}rvi and Mervi Toivari",

note = "Publisher Copyright: {\textcopyright} The Author(s) 2021. Published by Oxford University Press on behalf of Society of Industrial Microbiology and Biotechnology.",

year = "2021",

month = jun,

day = "1",

doi = "10.1093/jimb/kuab028",

language = "English",

volume = "48",

journal = "Journal of industrial microbiology and biotechnology",

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TY - JOUR

T1 - Production of D-lactic acid containing polyhydroxyalkanoate polymers in yeast Saccharomyces cerevisiae

AU - Ylinen, Anna

AU - Maaheimo, Hannu

AU - Anghelescu-Hakala, Adina

AU - Penttilä, Merja

AU - Salusjärvi, Laura

AU - Toivari, Mervi

N1 - Publisher Copyright: © The Author(s) 2021. Published by Oxford University Press on behalf of Society of Industrial Microbiology and Biotechnology.

PY - 2021/6/1

Y1 - 2021/6/1

N2 - 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 PhaC1437Ps6-19, propionyl-CoA transferase Pct540Cp, acetyl-CoA acetyltransferase 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.

AB - 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 PhaC1437Ps6-19, propionyl-CoA transferase Pct540Cp, acetyl-CoA acetyltransferase 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.

KW - Biopolymer

KW - Copolymer

KW - PHB

KW - Poly(D-lactic acid)

KW - Saccharomyces cerevisiae

UR - https://www.scopus.com/pages/publications/85111790197

U2 - 10.1093/jimb/kuab028

DO - 10.1093/jimb/kuab028

M3 - Article

C2 - 33899921

SN - 1367-5435

VL - 48

JO - Journal of industrial microbiology and biotechnology

JF - Journal of industrial microbiology and biotechnology

IS - 5-6

M1 - kuab028

ER -

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

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