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 - Funding Information:
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.
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 - http://www.scopus.com/inward/record.url?scp=85111790197&partnerID=8YFLogxK
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 -