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Engineering of Saccharomyces cerevisiae for the production of poly-3-d-hydroxybutyrate from xylose.

Sandström AG, Muñoz de Las Heras A, Portugal-Nunes D, Gorwa-Grauslund MF - AMB Express (2015)

Bottom Line: In the present study, the robust baker's yeast Saccharomyces cerevisiae was engineered to produce PHB from xylose, the main pentose found in lignocellulosic biomass.The PHB pathway genes from the well-characterized PHB producer Cupriavidus necator were introduced in recombinant S. cerevisiae strains already capable of pentose utilization by introduction of the fungal genes for xylose utilization from the yeast Scheffersomyces stipitis.PHB production from xylose was successfully demonstrated in shake-flasks experiments, with PHB yield of 1.17 ± 0.18 mg PHB g(-1) xylose.

View Article: PubMed Central - PubMed

Affiliation: Division of Applied Microbiology, Department of Chemistry, Lund University, PO Box 124, Lund, SE-221 00 Sweden.

ABSTRACT
Poly-3-d-hydroxybutyrate (PHB) is a promising biopolymer naturally produced by several bacterial species. In the present study, the robust baker's yeast Saccharomyces cerevisiae was engineered to produce PHB from xylose, the main pentose found in lignocellulosic biomass. The PHB pathway genes from the well-characterized PHB producer Cupriavidus necator were introduced in recombinant S. cerevisiae strains already capable of pentose utilization by introduction of the fungal genes for xylose utilization from the yeast Scheffersomyces stipitis. PHB production from xylose was successfully demonstrated in shake-flasks experiments, with PHB yield of 1.17 ± 0.18 mg PHB g(-1) xylose. Under well-controlled fully aerobic conditions, a titer of 101.7 mg PHB L(-1) was reached within 48 hours, with a PHB yield of 1.99 ± 0.15 mg PHB g(-1) xylose, thereby demonstrating the potential of this host for PHB production from lignocellulose.

No MeSH data available.


Related in: MedlinePlus

The PHB metabolic pathway, consisting of A) metabolite structures, involved enzymes, cofactors and B) the genes and regulatory sequences in the integrative vector YIpAGS2 in a schematic representation.
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Fig1: The PHB metabolic pathway, consisting of A) metabolite structures, involved enzymes, cofactors and B) the genes and regulatory sequences in the integrative vector YIpAGS2 in a schematic representation.

Mentions: The C. necator PHB pathway, consisting of a β-ketothiolase (acetyl-CoA acetyltransferase) (encoded by PhaA), an acetoacetyl-CoA reductase (encoded by PhaB1), and a PHB synthase (encoded by PhaC1) (Figure 1A), was chosen for PHB production in S. cerevisiae as it has been successfully used in the past (see e.g. (Carlson and Srienc 2006; Kocharin et al. 2012; Breuer et al. 2002)). The genes, ordered from MWG Eurofins, were codon optimized for S. cerevisiae, which notably led to lower GC-content (47–52% GC) than the native bacterial genes (63–68% GC). All three genes were cloned behind a strong constitutive promoter (pTEF1, pTPI1 and pGPM1 for PhaA, PhaB1 and PhaC1, respectively) into a single large vector, named YIpAGS2 (Figure 1B), based on YIplac128 which has a LEU2 gene for complementation of auxotrophic strains. The entire pathway was sequenced and validated.Figure 1


Engineering of Saccharomyces cerevisiae for the production of poly-3-d-hydroxybutyrate from xylose.

Sandström AG, Muñoz de Las Heras A, Portugal-Nunes D, Gorwa-Grauslund MF - AMB Express (2015)

The PHB metabolic pathway, consisting of A) metabolite structures, involved enzymes, cofactors and B) the genes and regulatory sequences in the integrative vector YIpAGS2 in a schematic representation.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4385036&req=5

Fig1: The PHB metabolic pathway, consisting of A) metabolite structures, involved enzymes, cofactors and B) the genes and regulatory sequences in the integrative vector YIpAGS2 in a schematic representation.
Mentions: The C. necator PHB pathway, consisting of a β-ketothiolase (acetyl-CoA acetyltransferase) (encoded by PhaA), an acetoacetyl-CoA reductase (encoded by PhaB1), and a PHB synthase (encoded by PhaC1) (Figure 1A), was chosen for PHB production in S. cerevisiae as it has been successfully used in the past (see e.g. (Carlson and Srienc 2006; Kocharin et al. 2012; Breuer et al. 2002)). The genes, ordered from MWG Eurofins, were codon optimized for S. cerevisiae, which notably led to lower GC-content (47–52% GC) than the native bacterial genes (63–68% GC). All three genes were cloned behind a strong constitutive promoter (pTEF1, pTPI1 and pGPM1 for PhaA, PhaB1 and PhaC1, respectively) into a single large vector, named YIpAGS2 (Figure 1B), based on YIplac128 which has a LEU2 gene for complementation of auxotrophic strains. The entire pathway was sequenced and validated.Figure 1

Bottom Line: In the present study, the robust baker's yeast Saccharomyces cerevisiae was engineered to produce PHB from xylose, the main pentose found in lignocellulosic biomass.The PHB pathway genes from the well-characterized PHB producer Cupriavidus necator were introduced in recombinant S. cerevisiae strains already capable of pentose utilization by introduction of the fungal genes for xylose utilization from the yeast Scheffersomyces stipitis.PHB production from xylose was successfully demonstrated in shake-flasks experiments, with PHB yield of 1.17 ± 0.18 mg PHB g(-1) xylose.

View Article: PubMed Central - PubMed

Affiliation: Division of Applied Microbiology, Department of Chemistry, Lund University, PO Box 124, Lund, SE-221 00 Sweden.

ABSTRACT
Poly-3-d-hydroxybutyrate (PHB) is a promising biopolymer naturally produced by several bacterial species. In the present study, the robust baker's yeast Saccharomyces cerevisiae was engineered to produce PHB from xylose, the main pentose found in lignocellulosic biomass. The PHB pathway genes from the well-characterized PHB producer Cupriavidus necator were introduced in recombinant S. cerevisiae strains already capable of pentose utilization by introduction of the fungal genes for xylose utilization from the yeast Scheffersomyces stipitis. PHB production from xylose was successfully demonstrated in shake-flasks experiments, with PHB yield of 1.17 ± 0.18 mg PHB g(-1) xylose. Under well-controlled fully aerobic conditions, a titer of 101.7 mg PHB L(-1) was reached within 48 hours, with a PHB yield of 1.99 ± 0.15 mg PHB g(-1) xylose, thereby demonstrating the potential of this host for PHB production from lignocellulose.

No MeSH data available.


Related in: MedlinePlus