Genetic engineering of Synechocystis PCC6803 for the photoautotrophic production of the sweetener erythritol

Woude, Aniek D. van der; Gallego, Ruth Pérez; Vreugdenhil, Angie; Veetil, Vinod Puthan; Chroumpi, Tania; Hellingwerf, Klaas J.

doi:10.1186/s12934-016-0458-y

Cited by 33 publications

(17 citation statements)

References 36 publications

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“…Recently, Janek et al [ 15 ] identified an ylER enzyme ( YALI0F18590p ) in Y. lipolytica based on sequence similarity with erythrose reductase from Candida magnoliae [ 7 ]. These ER enzymes, belonging to the aldose reductase family (ALR), have also been reported mainly dependent on NADPH as a redox co-factor [ 5 , 18 , 21 , 22 ]. Different ER isozymes have been reported in T. megachiliensis [ 18 ] and Moniliella sp.…”

Section: Resultsmentioning

confidence: 99%

Identification, characterization of two NADPH-dependent erythrose reductases in the yeast Yarrowia lipolytica and improvement of erythritol productivity using metabolic engineering

et al. 2018

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BackgroundErythritol is a four-carbon sugar alcohol with sweetening properties that is used by the agro-food industry as a food additive. In the yeast Yarrowia lipolytica, the last step of erythritol synthesis involves the reduction of erythrose by specific erythrose reductase(s). In the earlier report, an erythrose reductase gene (YALI0F18590g) from erythritol-producing yeast Y. lipolytica MK1 was identified (Janek et al. in Microb Cell Fact 16:118, 2017). However, deletion of the gene in Y. lipolytica MK1 only resulted in some lower erythritol production but the erythritol synthesis process was still maintained, indicating that other erythrose reductase gene(s) might exist in the genome of Y. lipolytica.ResultsIn this study, we have isolated genes g141.t1 (YALI0D07634g) and g3023.t1 (YALI0C13508g) encoding two novel erythrose reductases (ER). The biochemical characterization of the purified enzymes showed that they have a strong affinity for erythrose. Deletion of the two ER genes plus g801.t1 (YALI0F18590g) did not prevent erythritol synthesis, suggesting that other ER or ER-like enzymes remain to be discovered in this yeast. Overexpression of the newly isolated two genes (ER10 or ER25) led to an average 14.7% higher erythritol yield and 31.2% higher productivity compared to the wild-type strain. Finally, engineering NADPH cofactor metabolism by overexpression of genes ZWF1 and GND1 encoding glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase, respectively, allowed a 23.5% higher erythritol yield and 50% higher productivity compared to the wild-type strain. The best of our constructed strains produced an erythritol titer of 190 g/L in baffled flasks using glucose as main carbon source.ConclusionsOur results highlight that in the Y. lipolytica genome several genes encode enzymes able to reduce erythrose into erythritol. The catalytic properties of these enzymes and their cofactor dependency are different from that of already known erythrose reductase of Y. lipolytica. Constitutive expression of the newly isolated genes and engineering of NADPH cofactor metabolism led to an increase in erythritol titer. Development of fermentation strategies will allow further improvement of this productivity in the future.Electronic supplementary materialThe online version of this article (10.1186/s12934-018-0982-z) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Identification, characterization of two NADPH-dependent erythrose reductases in the yeast Yarrowia lipolytica and improvement of erythritol productivity using metabolic engineering

et al. 2018

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“…van der Woude et al. aimed to improve erythritol yield by overexpressing phosphoketolase or TK gene and consequently increase erythrose‐4‐P availability in the metabolic pathway of erythritol in Synechocystis PCC6803, while minimal effects were observed. Bellou et al.…”

Section: Resultsmentioning

confidence: 99%

“…According to the study of Ghezelbash et al [35], no remarkable difference was observed in ER gene expression levels between the wild-type Y. lipolytica and mutant strains, and the specific activity of ER was probably due to the modification of the active center of the enzyme. van der Woude et al [36] aimed to improve erythritol yield by overexpressing phosphoketolase or TK gene and consequently increase erythrose-4-P availability in the metabolic pathway of erythritol in Synechocystis PCC6803, while minimal effects were observed. Bellou et al [37] reported that the activities of the key enzymes involved in lipid synthesis by Y. lipolytica were influenced at post-transcriptional level rather than mRNA expression level when cultivated under nitrogen-and magnesium-limited conditions.…”

Section: Expression Levels Of Relative Enzymes In M53mentioning

confidence: 99%

Effects of osmotic pressure and pH on citric acid and erythritol production from waste cooking oil by Yarrowia lipolytica

Liu

et al. 2018

Engineering in Life Sciences

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Erythritol and citric acid could be produced from waste cooking oil (WCO) by Yarrowia lipolytica under different medium conditions, and osmotic pressure together with pH were considered to be the critical factors in this process. High osmotic pressure (2.76 osmol/L) combined with low pH (pH 3.0) promoted the highest yield of erythritol (21.8 g/L) accompanied by low‐producing citric acid (2.5 g/L). By contrast, the highest citric acid biosynthesis (12.6 g/L) was detected under a pH of 6.0 and an osmotic pressure of 0.75 osmol/L, when only 4.0 g/L of erythritol was yielded. Moreover, lipase activities in these two media were also detected, and pH 3.0–OP 2.76 was supposed to be more beneficial to lipase activity. Biochemical pathways involved in the biosynthesis of erythritol and citric acid were subsequently investigated, and the products yielded from WCO were assumed to be correlated with the activities of transketolase, erythrose reductase, citrate synthase, and glycerol kinase. However, RT‐PCR analysis revealed that mRNA levels of these enzymes did not significantly differ, confirming that metabolic flux regulations of erythritol and citric acid mostly took place at the post‐transcriptional level.

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“…The expressed enzymes can then convert erythrose-4-phosphate into erythritol. Erythrose-4-phosphate is inherent and is built as a key intermediate of the CO 2 -fixing Calvin Benson Bassham cycle (van der Woude et al 2016 ).…”

Section: Production—history and Developmentmentioning

confidence: 99%

Erythritol as sweetener—wherefrom and whereto?

Regnat

Mach

Mach-Aigner

2017

Appl Microbiol Biotechnol

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Erythritol is a naturally abundant sweetener gaining more and more importance especially within the food industry. It is widely used as sweetener in calorie-reduced food, candies, or bakery products. In research focusing on sugar alternatives, erythritol is a key issue due to its, compared to other polyols, challenging production. It cannot be chemically synthesized in a commercially worthwhile way resulting in a switch to biotechnological production. In this area, research efforts have been made to improve concentration, productivity, and yield. This mini review will give an overview on the attempts to improve erythritol production as well as their development over time.

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Genetic engineering of Synechocystis PCC6803 for the photoautotrophic production of the sweetener erythritol

Cited by 33 publications

References 36 publications

Identification, characterization of two NADPH-dependent erythrose reductases in the yeast Yarrowia lipolytica and improvement of erythritol productivity using metabolic engineering

Identification, characterization of two NADPH-dependent erythrose reductases in the yeast Yarrowia lipolytica and improvement of erythritol productivity using metabolic engineering

Effects of osmotic pressure and pH on citric acid and erythritol production from waste cooking oil by Yarrowia lipolytica

Erythritol as sweetener—wherefrom and whereto?

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