Highly Efficient Fructooligosaccharides Production by an Erythritol-Producing Yeast <i>Yarrowia lipolytica</i> Displaying Fructosyltransferase

Zhang, Lebin; An, Jiae; Li, Lijuan; Wang, Hengwei; Liu, Dawen; Li, Ning; Cheng, Hairong; Deng, Zixin

doi:10.1021/acs.jafc.6b00115

Cited by 35 publications

(21 citation statements)

References 42 publications

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“…To construct the ER10 disruption cassette, 2 kb DNA fragment located upstream (P) and downstream (T) of gene g141.t1 were PCR amplified using genomic DNA of strain CGMCC7326 as a template, and primers pairs ER10-F1/ER10-R1 and ER10-F2/ER10-R2, respectively. After purification, P fragment was digested with Not I and Sal I and cloned at corresponding sites of plasmid pINA-Pir1-A.oryFTase [ 38 ], to yield plasmid pINA-UP ER10 (Additional file 1 : Table S1). The purified T fragment was then digested with Eco RI and Not I, and cloned at corresponding sites of plasmid pINA-UP ER10 , to yield plasmid pINA-UP ER10 -DW ER10 .…”

Section: Methodsmentioning

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: Methodsmentioning

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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“…Under the optimal conditions, the wholecell catalyst produced 480 g/L FOS from 800 g/L sucrose, with a yield of 60% and productivity of 160 g/(L•h). The engineered yeast pastes from erythritol industry was stable in FOS production, with only 10% FTase activity lost after 10 recycling number (Zhang et al, 2016). FOSs can also be produced by directly hydrolyzing inulin by the endo-inulinase (EC 3.2.1.7) (Singh et al, 2016).…”

Section: Fructo-oligosaccharidesmentioning

confidence: 99%

Recent Advances in Producing Sugar Alcohols and Functional Sugars by Engineering Yarrowia lipolytica

Abbasi

Liu

Zhi

et al. 2021

Front. Bioeng. Biotechnol.

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The sugar alcohols and functional sugars have wide applications in food, pharmaceutical, and chemical industries. However, the smaller quantities of natural occurring sugar alcohols and functional sugars restricted their applications. The enzymatic and whole-cell catalyst production is emerging as the predominant alternatives. The properties of Yarrowia lipolytica make it a promising sugar alcohol and functional sugar producer. However, there are still some issues to be resolved. As there exist reviews about the chemical structures, physicochemical properties, biological functions, applications, and biosynthesis of sugar alcohols and/or functional sugars in Y. lipolytica, this mini review will not only update the recent advances in enzymatic and microbial production of sugar alcohols (erythritol, D-threitol, and xylitol) and functional sugars (isomaltulose, trehalose, fructo-oligosaccharides, and galacto-oligosaccharides) by using recombinant Y. lipolytica but also focus on the studies of gene discovery, pathway engineering, expanding substrate scope, bioprocess engineering, and novel breeding methods to resolve the aforementioned issues.

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“…Zhang et al. 53 displayed fructosyltransferase from A. oryzae on the cell surface of Yarrowia lipolytica , and the FOS yield reached 480 g L −1 .…”

Section: Production Of Fructosyltransferasesmentioning

confidence: 99%

Research Progress and Prospects for Fructosyltransferases

et al. 2021

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Fructosyltransferases can be used to produce fructooligosaccharides from sucrose and are obtained from different sources, mostly microorganisms and plants. Heterologous expression and several mutagenesis methods were applied to improve the production of fructosyltransferases. The structures and characteristics of fructosyltransferases were investigated. Immobilization and protein engineering can improve their characteristics. In this review, screening and mutagenesis of microorganisms, heterologous expression, purification, characterization, structural analysis, immobilization, and protein engineering of fructosyltransferases are discussed. Future prospects in this field are also considered.

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Highly Efficient Fructooligosaccharides Production by an Erythritol-Producing Yeast Yarrowia lipolytica Displaying Fructosyltransferase

Cited by 35 publications

References 42 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

Recent Advances in Producing Sugar Alcohols and Functional Sugars by Engineering Yarrowia lipolytica

Research Progress and Prospects for Fructosyltransferases

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