In <i>Yarrowia lipolytica</i> erythritol catabolism ends with erythrose phosphate

Niang, Papa Makhona; Arias, Anthony Argüelles; Steels, Sébastien; Denies, Olivia; Nicaud, Jean‐Marc; Fickers, Patrick

doi:10.1002/cbin.11265

Cited by 17 publications

(7 citation statements)

References 24 publications

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“…Although Y. lipolytica can synthesize erythritol in response to osmotic stress, it also has the ability to reconsume it in isotonic conditions. In this catabolic pathway, erythritol is first converted into erythrulose by an erythritol dehydrogenase encoded by gene EYD1 [26]. Therefore, the gene was disrupted in strains HCY109-2 (△ku70△ura3) and HCY113 (△ku70△ura3△mdh2).…”

Section: Disruption Of Yleyd1mentioning

confidence: 99%

Metabolic engineering of Yarrowia lipolytica for thermoresistance and enhanced erythritol productivity

Wang

Chi

Zou

et al. 2020

Biotechnol Biofuels

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Background Functional sugar alcohols have been widely used in the food, medicine, and pharmaceutical industries for their unique properties. Among these, erythritol is a zero calories sweetener produced by the yeast Yarrowia lipolytica. However, in wild-type strains, erythritol is produced with low productivity and yield and only under high osmotic pressure together with other undesired polyols, such as mannitol or d-arabitol. The yeast is also able to catabolize erythritol in non-stressing conditions. Results Herein, Y. lipolytica has been metabolically engineered to increase erythritol production titer, yield, and productivity from glucose. This consisted of the disruption of anabolic pathways for mannitol and d-arabitol together with the erythritol catabolic pathway. Genes ZWF1 and GND encoding, respectively, glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase were also constitutively expressed in regenerating the NADPH2 consumed during erythritol synthesis. Finally, the gene RSP5 gene from Saccharomyces cerevisiae encoding ubiquitin ligase was overexpressed to improve cell thermoresistance. The resulting strain HCY118 is impaired in mannitol or d-arabitol production and erythritol consumption. It can grow well up to 35 °C and retain an efficient erythritol production capacity at 33 °C. The yield, production, and productivity reached 0.63 g/g, 190 g/L, and 1.97 g/L·h in 2-L flasks, and increased to 0.65 g/g, 196 g/L, and 2.51 g/L·h in 30-m3 fermentor, respectively, which has economical practical importance. Conclusion The strategy developed herein yielded an engineered Y. lipolytica strain with enhanced thermoresistance and NADPH supply, resulting in a higher ability to produce erythritol, but not mannitol or d-arabitol from glucose. This is of interest for process development since it will reduce the cost of bioreactor cooling and erythritol purification.

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Section: Disruption Of Yleyd1mentioning

confidence: 99%

Metabolic engineering of Yarrowia lipolytica for thermoresistance and enhanced erythritol productivity

Wang

Chi

Zou

et al. 2020

Biotechnol Biofuels

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“…In Y. lipolytica, three genes encoding erythrose reductase have been characterized so far (namely, YALI0F18590g, YALI0D07634g and YALI0C13508g) [99,100]. We also recently elucidated the full EOL catabolic pathway [101]. It is first converted into erythrulose by an erythritol dehydrogenase (EYD1, YALI0F01650g) and subsequently phosphorylated into L-erythrulose-1P by an erythrulose kinase (EYK1, YALI0F01606g).…”

Section: Erythritolmentioning

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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“…It has been suggested that EUF1 (YALI0F01562g) would code for a transcription factor involved in erythritol utilization [ 28 ]. All five genes mentioned above are located in Y. lipolytica ’s chromosome F in a region called the erythritol utilization cluster [ 29 , 30 ] (Fig. 1 a), and their expression can be induced by erythritol as well as by erythrulose.…”

Section: Introductionmentioning

confidence: 99%

Bidirectional hybrid erythritol-inducible promoter for synthetic biology in Yarrowia lipolytica

et al. 2023

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Background The oleaginous yeast Yarrowia lipolytica is increasingly used as a chassis strain for generating bioproducts. Several hybrid promoters with different strengths have been developed by combining multiple copies of an upstream activating sequence (UAS) associated with a TATA box and a core promoter. These promoters display either constitutive, phase-dependent, or inducible strong expression. However, there remains a lack of bidirectional inducible promoters for co-expressing genes in Y. lipolytica. Results This study built on our previous work isolating and characterizing the UAS of the erythritol-induced genes EYK1 and EYD1 (UAS-eyk1). We found an erythritol-inducible bidirectional promoter (BDP) located in the EYK1-EYL1 intergenic region. We used the BDP to co-produce YFP and RedStarII fluorescent proteins and demonstrated that the promoter’s strength was 2.7 to 3.5-fold stronger in the EYL1 orientation compared to the EYK1 orientation. We developed a hybrid erythritol-inducible bidirectional promoter (HBDP) containing five copies of UAS-eyk1 in both orientations. It led to expression levels 8.6 to 19.2-fold higher than the native bidirectional promoter. While the BDP had a twofold-lower expression level than the strong constitutive TEF promoter, the HBDP had a 5.0-fold higher expression level when oriented toward EYL1 and a 2.4-fold higher expression level when oriented toward EYK1. We identified the optimal media for BDP usage by exploring yeast growth under microbioreactor conditions. Additionally, we constructed novel Golden Gate biobricks and a destination vector for general use. Conclusions In this research, we developed novel bidirectional and hybrid bidirectional promoters of which expression can be fine-tuned, responding to the need for versatile promoters in the yeast Y. lipolytica. This study provides effective tools that can be employed to smoothly adjust the erythritol-inducible co-expression of two target genes in biotechnology applications. BDPs developed in this study have potential applications in the fields of heterologous protein production, metabolic engineering, and synthetic biology.

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In Yarrowia lipolytica erythritol catabolism ends with erythrose phosphate

Cited by 17 publications

References 24 publications

Metabolic engineering of Yarrowia lipolytica for thermoresistance and enhanced erythritol productivity

Metabolic engineering of Yarrowia lipolytica for thermoresistance and enhanced erythritol productivity

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

Bidirectional hybrid erythritol-inducible promoter for synthetic biology in Yarrowia lipolytica

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