Polyhydroxybutyrate (PHB) Biosynthesis by an Engineered <i>Yarrowia lipolytica</i> Strain Using Co-Substrate Strategy
High production cost is one of the major factors that limit the market growth of <i>polyhydroxyalkanoates</i> (PHAs) as a biopolymer. Improving PHA synthesis performance and utilizing low-grade feedstocks are two logical strategies for reducing costs. As an oleaginous yeast, <i>Y....
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-11-01
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| Series: | Fermentation |
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| Online Access: | https://www.mdpi.com/2311-5637/9/12/1003 |
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| author | Masoud Tourang Xiaochao Xiong Sara Sarkhosh Shulin Chen |
| author_facet | Masoud Tourang Xiaochao Xiong Sara Sarkhosh Shulin Chen |
| author_sort | Masoud Tourang |
| collection | DOAJ |
| description | High production cost is one of the major factors that limit the market growth of <i>polyhydroxyalkanoates</i> (PHAs) as a biopolymer. Improving PHA synthesis performance and utilizing low-grade feedstocks are two logical strategies for reducing costs. As an oleaginous yeast, <i>Y. lipolytica</i> has a high carbon flux through acetyl-CoA (the main PHB precursor), which makes it a desired cell factory for PHB biosynthesis. In the current study, two different metabolic pathways (<i>NBC</i> and <i>ABC</i>) were introduced into <i>Y. lipolytica PO1f</i> for synthesizing PHB. Compared to the <i>ABC</i> pathway, the <i>NBC</i> pathway, which includes <i>NphT7</i> to redirect the lipogenesis pathway and catalyze acetoacetyl-CoA synthesis in a more energy-favored reaction, led to PHB accumulation of up to 11% of cell dry weight (CDW), whereas the <i>ABC</i> pathway resulted in non-detectable accumulations of PHB. Further modifications of the strain with the <i>NBC</i> pathway through peroxisomal compartmentalization and gene dose overexpression reached 41% PHB of CDW and a growth rate of 0.227 h<sup>−1</sup>. A low growth rate was observed with acetate as the sole source of carbon and energy or glucose as the sole substrate at high concentrations. Using a co-substrate strategy helped overcoming the inhibitory and toxic effects of both substrates. Cultivating the engineered strain in the optimal co-substrate condition predicted by response surface methodology (<i>RSM</i>) led to 83.4 g/L of biomass concentration and 31.7 g/L of PHB. These results offer insight into a more cost-effective production of PHB with engineered <i>Y. lipolytica</i>. |
| first_indexed | 2024-03-08T20:46:50Z |
| format | Article |
| id | doaj.art-7396d9f73a4e4043899916a31f4e7692 |
| institution | Directory Open Access Journal |
| issn | 2311-5637 |
| language | English |
| last_indexed | 2024-03-08T20:46:50Z |
| publishDate | 2023-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Fermentation |
| spelling | doaj.art-7396d9f73a4e4043899916a31f4e76922023-12-22T14:07:51ZengMDPI AGFermentation2311-56372023-11-01912100310.3390/fermentation9121003Polyhydroxybutyrate (PHB) Biosynthesis by an Engineered <i>Yarrowia lipolytica</i> Strain Using Co-Substrate StrategyMasoud Tourang0Xiaochao Xiong1Sara Sarkhosh2Shulin Chen3Department of Biological Systems Engineering, Washington State University, L. J. Smith Hall, Pullman, WA 99164, USADepartment of Biological Systems Engineering, Washington State University, L. J. Smith Hall, Pullman, WA 99164, USADepartment of Biological Systems Engineering, Washington State University, L. J. Smith Hall, Pullman, WA 99164, USADepartment of Biological Systems Engineering, Washington State University, L. J. Smith Hall, Pullman, WA 99164, USAHigh production cost is one of the major factors that limit the market growth of <i>polyhydroxyalkanoates</i> (PHAs) as a biopolymer. Improving PHA synthesis performance and utilizing low-grade feedstocks are two logical strategies for reducing costs. As an oleaginous yeast, <i>Y. lipolytica</i> has a high carbon flux through acetyl-CoA (the main PHB precursor), which makes it a desired cell factory for PHB biosynthesis. In the current study, two different metabolic pathways (<i>NBC</i> and <i>ABC</i>) were introduced into <i>Y. lipolytica PO1f</i> for synthesizing PHB. Compared to the <i>ABC</i> pathway, the <i>NBC</i> pathway, which includes <i>NphT7</i> to redirect the lipogenesis pathway and catalyze acetoacetyl-CoA synthesis in a more energy-favored reaction, led to PHB accumulation of up to 11% of cell dry weight (CDW), whereas the <i>ABC</i> pathway resulted in non-detectable accumulations of PHB. Further modifications of the strain with the <i>NBC</i> pathway through peroxisomal compartmentalization and gene dose overexpression reached 41% PHB of CDW and a growth rate of 0.227 h<sup>−1</sup>. A low growth rate was observed with acetate as the sole source of carbon and energy or glucose as the sole substrate at high concentrations. Using a co-substrate strategy helped overcoming the inhibitory and toxic effects of both substrates. Cultivating the engineered strain in the optimal co-substrate condition predicted by response surface methodology (<i>RSM</i>) led to 83.4 g/L of biomass concentration and 31.7 g/L of PHB. These results offer insight into a more cost-effective production of PHB with engineered <i>Y. lipolytica</i>.https://www.mdpi.com/2311-5637/9/12/1003acetategene dosage effectacetoacetyl-CoA synthasecompartmentalizationsubstrate inhibition |
| spellingShingle | Masoud Tourang Xiaochao Xiong Sara Sarkhosh Shulin Chen Polyhydroxybutyrate (PHB) Biosynthesis by an Engineered <i>Yarrowia lipolytica</i> Strain Using Co-Substrate Strategy Fermentation acetate gene dosage effect acetoacetyl-CoA synthase compartmentalization substrate inhibition |
| title | Polyhydroxybutyrate (PHB) Biosynthesis by an Engineered <i>Yarrowia lipolytica</i> Strain Using Co-Substrate Strategy |
| title_full | Polyhydroxybutyrate (PHB) Biosynthesis by an Engineered <i>Yarrowia lipolytica</i> Strain Using Co-Substrate Strategy |
| title_fullStr | Polyhydroxybutyrate (PHB) Biosynthesis by an Engineered <i>Yarrowia lipolytica</i> Strain Using Co-Substrate Strategy |
| title_full_unstemmed | Polyhydroxybutyrate (PHB) Biosynthesis by an Engineered <i>Yarrowia lipolytica</i> Strain Using Co-Substrate Strategy |
| title_short | Polyhydroxybutyrate (PHB) Biosynthesis by an Engineered <i>Yarrowia lipolytica</i> Strain Using Co-Substrate Strategy |
| title_sort | polyhydroxybutyrate phb biosynthesis by an engineered i yarrowia lipolytica i strain using co substrate strategy |
| topic | acetate gene dosage effect acetoacetyl-CoA synthase compartmentalization substrate inhibition |
| url | https://www.mdpi.com/2311-5637/9/12/1003 |
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