Restoring of Glucose Metabolism of Engineered <i>Yarrowia lipolytica</i> for Succinic Acid Production via a Simple and Efficient Adaptive Evolution Strategy

Yang, Xiaofeng; Wang, Huaimin; Li, Chong; Lin, Carol Sze Ki

doi:10.1021/acs.jafc.7b00519

Cited by 60 publications

(34 citation statements)

References 41 publications

(96 reference statements)

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“…Gao et al inactivated sdh5 encoding succinate dehydrogenase assembly factor 2 (YALI0F11957g) in Po1f strain (derived from W29 strain) and obtained a mutant PGC01003 [23], and this strain showed impaired growth on glucose. The PGC01003 strain was subjected to adaptive evolution using glucose-based medium for 21 days and the evolved strain was designated as Y. lipolytica PSA02004 [18]. This strain was cultured on glucose, glycerol, xylose, glucose/xylose and glycerol/xylose (Figure 2).…”

Section: Resultsmentioning

confidence: 99%

Bioproduction of succinic acid from xylose by engineered Yarrowia lipolytica without pH control

Prabhu

Amaro

Lin

et al. 2020

Preprint

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Background : Xylose is a most prevalent sugar available in hemicellulose fraction of lignocellulosic biomass (LCB) and of great interest for the green economy. Unfortunately, most of the cell factories cannot inherently metabolize xylose as sole carbon source. Yarrowia lipolytica is a non-conventional yeast to produce industrially important metabolites. The yeast is able to metabolize a large variety of substrates including both hydrophilic and hydrophobic carbon sources. However, Y. lipolytica lacks effective metabolic pathway for xylose uptake and only scarce information is available on utilization of xylose. For the economically feasibility of LCB-based biorefineries, effective utilization of both pentose and hexose sugars is obligatory. Results : In the present study, succinic acid (SA) production from xylose by Y. lipolytica was examined. To this end, Y. lipolytica PSA02004 strain was engineered by overexpressing pentose pathway cassette comprising of xylose reductase ( XR ), xylitol dehydrogenase ( XDH ) and xylulose kinase ( XK ) gene. The recombinant strain exhibited a robust growth on xylose as sole carbon source and produced substantial amount of SA. The inhibition of cell growth and SA formation was observed above 60 g/L xylose concentration. The batch cultivation of recombinant strain in bioreactor resulted in a maximum biomass concentration of 7.3 g/L and SA titer of 11.2 g/L with the yield of 0.19 g/g. Similar results in term of cell growth and SA production were obtained with xylose-rich hydrolysate derived from sugarcane bagasse. The fed-batch fermentation yielded biomass concentration of 11.8 g/L (OD 600 : 56.1) and SA titer of 22.3 g/L with a gradual decrease in pH below 4.0. Acetic acid was obtained as a main byproduct in all the fermentations. Conclusion : The recombinant strain displayed potential for bioconversion of xylose to SA. Further, this study provided a new insight on conversion of lignocellulosic biomass into value-added products. To the best of our knowledge, this is the first study on SA production by Y. lipolytica using xylose as a sole carbon source.

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Section: Resultsmentioning

confidence: 99%

Bioproduction of succinic acid from xylose by engineered Yarrowia lipolytica without pH control

Prabhu

Amaro

Lin

et al. 2020

Preprint

Self Cite

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“…Similar to the Y. lipolytica engineered strain Y-3314, strain PGC01003 has an impaired glucose metabolism. The strain underwent an adaptive evolution strategy at pH 6 with the aim to restore glucose metabolism [74]. After 21 days of evolution, growth ability on glucose was significantly increased, with a specific growth rate of 0.72 h −1 for the evolved strain PSA02004.…”

Section: Succinic Acidmentioning

confidence: 99%

Sugar Alcohols and Organic Acids Synthesis in Yarrowia lipolytica: Where Are We?

2020

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Sugar alcohols and organic acids that derive from the metabolism of certain microorganisms have a panoply of applications in agro-food, chemical and pharmaceutical industries. The main challenge in their production is to reach a productivity threshold that allow the process to be profitable. This relies on the construction of efficient cell factories by metabolic engineering and on the development of low-cost production processes by using industrial wastes or cheap and widely available raw materials as feedstock. The non-conventional yeast Yarrowia lipolytica has emerged recently as a potential producer of such metabolites owing its low nutritive requirements, its ability to grow at high cell densities in a bioreactor and ease of genome edition. This review will focus on current knowledge on the synthesis of the most important sugar alcohols and organic acids in Y. lipolytica.

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“…Gao et al inactivated sdh5 encoding succinate dehydrogenase assembly factor2 (YALI0F11957g) in Po1f strain (derived from W29 strain) and obtained a mutant PGC01003 [32], and this strain showed impaired growth on glucose. The PGC01003 strain was subjected to adaptive evolution using glucose-based medium for 21 days and the evolved strain was designated as Y. lipolytica PSA02004 [26]. This strain was cultured on glucose, glycerol, xylose, glucose/xylose and glycerol/xylose ( Figure 2).…”

Section: Shake Flask Cultivation Of Y Lipolytica Psa02004mentioning

confidence: 99%

“…This renders this yeast species as an attractive host to produce another TCA intermediate which is known as succinic acid. Previously, Y. lipolytica has been engineered for production succinic acid using glucose and glycerol as carbon sources [25,26]. Y. lipolytica strains can utilize a wide range of carbon source including glucose, glycerol, alkanes and lipids [27,28].…”

Section: Introductionmentioning

confidence: 99%

Bioproduction of succinic acid from xylose by engineered Yarrowia lipolytica without pH control

Prabhu

Amaro

Lin

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

Background Xylose is a most prevalent sugar available in hemicellulose fraction of lignocellulosic biomass (LCB) and of great interest for the green economy. Unfortunately, most of the cell factories cannot inherently metabolize xylose as sole carbon source. Yarrowia lipolytica is a non-conventional yeast to produce industrially important metabolites, and it is able to metabolize a large variety of substrates including both hydrophilic and hydrophobic carbon sources. However, Y. lipolytica lacks effective metabolic pathway for xylose uptake and only scarce information is available on utilization of xylose. For the economically feasible of LCB-based biorefineries, effective utilization of both pentose and hexose sugars is obligatory. Results In the present study, succinic acid (SA) production from xylose by Y. lipolytica was examined. To this end, Y. lipolytica PSA02004 strain was engineered by overexpressing pentose pathway cassette comprising of xylose reductase ( XR ), xylitol dehydrogenase ( XDH ) and xylulose kinase ( XK ) gene. The recombinant strain exhibited a robust growth on xylose as sole carbon source and accumulated SA (3.8 g/L) with a yield of 0.19 g/g in shake flask studies. Substrate inhibition studies revealed a marked negative impact on cell growth and product formation above 60 g/L xylose concentration. The modelling based on inhibition kinetics revealed that Aiba model showed better fit with experimental data, which resulted the correlation coefficient (R 2 ) of 0.82 and inhibition constant (K I ) 88.9 g/L. The batch cultivation of recombinant strain in bioreactor resulted in a maximum biomass concentration of 7.3 g/L and SA titer of 11.2 g/L with the yield of 0.18 g/g. Similar results in term of cell growth and SA production were obtained with xylose-rich hydrolysate derived from sugarcane bagasse. The fed-batch fermentation yielded biomass concentration of 11.8 g/L (OD 600 : 56.1) and SA titer of 22.3 g/L with a gradual decrease in pH below 4.0. Acetic acid was obtained as a main byproduct in all the fermentations. Conclusion The recombinant strain displayed potential bioconversion of xylose to succinic acid. Further this study provided a new insight on conversion of LCB into value-added products. To the best of our knowledge, this is the first study on SA production by Y. lipolytica using xylose as a sole carbon source.

show abstract

Restoring of Glucose Metabolism of Engineered Yarrowia lipolytica for Succinic Acid Production via a Simple and Efficient Adaptive Evolution Strategy

Cited by 60 publications

References 41 publications

Bioproduction of succinic acid from xylose by engineered Yarrowia lipolytica without pH control

Bioproduction of succinic acid from xylose by engineered Yarrowia lipolytica without pH control

Sugar Alcohols and Organic Acids Synthesis in Yarrowia lipolytica: Where Are We?

Bioproduction of succinic acid from xylose by engineered Yarrowia lipolytica without pH control

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