Highly Efficient Erythritol Recovery from Waste Erythritol Mother Liquor by a Yeast-Mediated Biorefinery Process

Wang, Siqi; Wang, Hengwei; Lv, Jiyang; Deng, Zixin; Cheng, Hairong

doi:10.1021/acs.jafc.7b04112

Cited by 16 publications

(8 citation statements)

References 37 publications

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“…Although Y. lipolytica is one of the best erythritol producers, it also presents some drawbacks such as the synthesis of byproducts including mannitol and d -arabitol. This negatively impacts the yield from glucose and complexifies the purification steps [ 9 , 13 , 19 – 22 ]. Therefore, we thought of constructing a strain unable to produce these byproducts.…”

Section: Resultsmentioning

confidence: 99%

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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: Resultsmentioning

confidence: 99%

“…Erythritol is predominantly produced from glucose by osmophilic yeast such as Candida magnoliae, Y. lipolytica, Torula sp., Moniliella megachiliensis, or Trichosporonoides oedocephalis [7][8][9][10]. Among these, Y. lipolytica is the most widely used for industrial erythritol production.…”

Section: Introductionmentioning

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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“…Morioka et al used this stream as eluent in chromatography separation (Morioka et al 2000 ). Wang et al proposed an approach consisting of the preliminary characterization of by-products in the residual mother liquor (by HPLC, TLC and GC-MS analytics), followed by the screening for a strain able to consume the residual polyols in the mother liquor except erythritol (Wang et al 2017b ). Thus, the yeast Candida maltose SJTU82 was found to consume glycerol, ribitol, arabitol, mannitol and residual glucose were in a fed-batch culture.…”

Section: Opportunities For the Production Of Erythritol In A Circular Economy Contextmentioning

confidence: 99%

From the culture broth to the erythritol crystals: an opportunity for circular economy

Daza-Serna

Serna-Loaiza

Masi

et al. 2021

Appl Microbiol Biotechnol

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The reduction of sugar intake by adults has been stated by the World Health Organization as an important strategy to reduce the risk of non-communicable diseases. Erythritol is a four-carbon sugar alcohol that is considered as a highly suitable substitution for sucrose. This review article covers approaches for the separate stages of the biotechnological production of erythritol from cultivation to the downstream section. The first part focuses on the cultivation stage and compares the yields of erythritol and arising by-products achieved with different types of substrates (commercial versus alternative ones). The reported numbers obtained with the most prominently used microorganisms in different cultivation methods (batch, fed-batch or continuous) are presented. The second part focuses on the downstream section and covers the applied technologies for cell removal, recovery, purification and concentration of erythritol crystals, namely centrifugation, membrane separation, ion and preparative chromatography, crystallization and drying. The final composition of the culture broth and the preparative chromatography separation performance were identified as critical points in the production of a high-purity erythritol fraction with a minimum amount of losses. During the review, the challenges for a biotechnological production of erythritol in a circular economy context are discussed, in particular regarding the usage of sustainable resources and minimizing waste streams. Key points • Substitution of sucrose by erythritol can be a step towards a healthier society • Biotechnological production of erythritol should follow a circular economy concept • Culture broth composition and preparative chromatography are keys for downstreaming • Substrate, mother liquor and nutrients are challenges for circular economy

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“…Recently, the yeast Yarrowia lipolytica has also been found to be an efficient erythritol producer [ 2 ]. Several processes based on wild-type strains have been developed [ 12 – 14 ], but the most promising processes are based on metabolically engineered strains. Overexpression of genes involved in PPP, namely transketolase ( TKL1 , YALI0E06479g), transaldolase ( TAL1 , YALI0F15587g) and erythrose reductase ( ylER , YALI0F18590g ), allowed the erythritol productivity to increase with various magnitudes (from 16 to 200%) [ 11 , 15 , 16 ].…”

Section: Introductionmentioning

confidence: 99%

“…In order to get more insights on ER in Y. lipolytica and with the aim to further increase the erythritol production, we identified and characterized two additional ER (namely ER10 and ER27) in Y. lipolytica strain CGMCC7326, a strain able to produce erythritol with very high titer (more than 150 g/L, [ 14 , 20 ]). Overexpression of those ER encoding genes together with ZWF1 ( YALI0E22649g ) and GND1 ( YALI0B15598g ) that allow NADPH to be replenished, yielded significantly improved erythritol productivity.…”

Section: Introductionmentioning

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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Highly Efficient Erythritol Recovery from Waste Erythritol Mother Liquor by a Yeast-Mediated Biorefinery Process

Cited by 16 publications

References 37 publications

Metabolic engineering of Yarrowia lipolytica for thermoresistance and enhanced erythritol productivity

Metabolic engineering of Yarrowia lipolytica for thermoresistance and enhanced erythritol productivity

From the culture broth to the erythritol crystals: an opportunity for circular economy

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

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