Multiple gene integration to promote erythritol production on glycerol in Yarrowia lipolytica

Zhang, Ling; Nie, Ming-Yue; Liu, Feng; Chen, Jun; Wei, Liujing; Hua, Qiang

doi:10.1007/s10529-021-03113-1

Cited by 19 publications

(7 citation statements)

References 35 publications

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“…lipolytica is conducive to the accumulation of lipids, erythritol, and lycopene, we overexpressed AMP deaminase (AMPD) and fructose-6-phosphate phosphoketolase, from Bifidobacterium breve (BbFPK), simultaneously in the strain Y. lipolytica ylmctg to accelerate the production of erythritol and increase the β-carotene production. , Overexpression of fructose-6-phosphate phosphoketolase (FPK) and phosphotransacetylase (PTA) pathways was beneficial to improve the production of acetyl-CoA and erythrose-4-phosphate in yeast. , However, there were no appreciable changes in erythritol or β-carotene production parameters (production, yield, and productivity) between strains ylmctg and ylmctgaf (Figure E, Table ). This is likely due to acetyl-CoA sufficiency for the downstream MVA pathway, and the contribution of AMPD is limited under the condition of coproduction of erythritol and β-carotene by the strain Y.…”

Section: Resultsmentioning

confidence: 99%

Concomitant Production of Erythritol and β-Carotene by Engineered Yarrowia lipolytica

Zhang

et al. 2023

J. Agric. Food Chem.

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While the expansion of the erythritol production industry has resulted in unprecedented production of yeast cells, it also suffers from a lack of effective utilization. β-Carotene is a value-added compound that can be synthesized by engineered Yarrowia lipolytica. Here, we first evaluated the production performance of erythritol-producing yeast strains under two different morphologies and then successfully constructed a chassis with yeast-like morphology by deleting Mhy1 and Cla4 genes. Subsequently, β-carotene synthesis pathway genes, CarRA and CarB from Blakeslea trispora, were introduced to construct the βcarotene and erythritol coproducing Y. lipolytica strain ylmcc. The rate-limiting genes GGS1 and tHMG1 were overexpressed to increase the β-carotene yield by 45.32-fold compared with the strain ylmcc. However, increased β-carotene accumulation led to prolonged fermentation time; therefore, transporter engineering through overexpression of YTH1 and YTH3 genes was used to alleviate fermentation delays. Using batch fermentation in a 3 L bioreactor, this engineered Y. lipolytica strain produced erythritol with production, yield, and productivity values of 171 g/L, 0.56 g/g glucose, and 2.38 g/(L•h), respectively, with a concomitant βcarotene yield of 47.36 ± 0.06 mg/g DCW. The approach presented here improves the value of erythritol-producing cells and offers a low-cost technique to obtain hydrophobic terpenoids.

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Section: Resultsmentioning

confidence: 99%

Concomitant Production of Erythritol and β-Carotene by Engineered Yarrowia lipolytica

Zhang

et al. 2023

J. Agric. Food Chem.

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“…This method enabled the integration of the β-carotene biosynthesis pathway into the rDNA locus of the Y. lipolytica chromosome with a reported maximum efficiency of 21%. Recently, Golden Gate assembly has significantly improved pathway assembly and construction efficiency, considered one of the most robust techniques for multi-gene assembly [ 38 , 39 ]. Therefore, Celinska et al [ 40 ] developed a versatile and robust DNA assembly platform for Y. lipolytica using the Golden Gate modular cloning.…”

Section: Gene Expression and Multi-gene Assemblymentioning

confidence: 99%

Advances and perspectives in genetic expression and operation for the oleaginous yeast Yarrowia lipolytica

Hu,

Ge,

Jiang

et al. 2024

Synthetic and Systems Biotechnology

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“…Recently, new studies have been published dealing with the optimization of processes for the production of citrate and erythritol from glycerol and the search for new strains with higher product titers [39]. As is well known, a good strategy for improving the properties of microorganisms is mutagenesis of pre-selected strains; nevertheless, this is a time-consuming and labor-intensive solution [40]. Given the available completely sequenced genome of Y. lipolytica, it became possible to identify and analyze genes at the molecular level [41].…”

Section: Introductionmentioning

confidence: 99%

Application of New Yarrowia lipolytica Transformants in Production of Citrates and Erythritol from Glycerol

Rywińska,

Tomaszewska-Hetman,

Lazar

et al. 2024

IJMS

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Citric acid and erythritol are obtained on an industrial scale using biotechnological methods. Due to the growing market demand for these products, research is underway to improve the process economics by introducing new microorganisms, in particular of the species Yarrowia lipolytica. The aim of this study was to evaluate transformants of Y. lipolytica for growth and ability to overproduce citric acids and erythritol from glycerol. The transformants were constructed by overexpressing glycerol kinase, methylcitrate synthase and mitochondrial succinate-fumarate transporter in the mutant Wratislavia 1.31. Next, strains were assessed for biosynthesis of citrate (pH 5.5; nitrogen limitation) and erythritol (pH 3.0; high osmotic pressure) from glycerol. Regardless of culture conditions strains, 1.31.GUT1/6 and 1.31.GUT1/6.CIT1/3 exhibited high rates of substrate utilization. Under conditions favoring citrate biosynthesis, both strains produced several percent more citrates, accompanied by higher erythritol production compared to the parental strain. During erythritol biosynthesis, the strain 1.31.GUT1/6.CIT1/3.E34672g obtained as a result of co-expression of all three genes stood out, producing 84.0 g/L of erythritol with yield and productivity of 0.54 g/g and 0.72 g/Lh, respectively, which places it in the group of the highest-ranked producers of erythritol among Y. lipolytica species.

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Multiple gene integration to promote erythritol production on glycerol in Yarrowia lipolytica

Cited by 19 publications

References 35 publications

Concomitant Production of Erythritol and β-Carotene by Engineered Yarrowia lipolytica

Concomitant Production of Erythritol and β-Carotene by Engineered Yarrowia lipolytica

Advances and perspectives in genetic expression and operation for the oleaginous yeast Yarrowia lipolytica

Application of New Yarrowia lipolytica Transformants in Production of Citrates and Erythritol from Glycerol

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