Molecular cloning and expression of fungal cellobiose transporters and β-glucosidases conferring efficient cellobiose fermentation in Saccharomyces cerevisiae

Bae, Yi-Hyun; Kang, Kyeong Hyeon; Jin, Yong Su; Seo, Jin Ho

doi:10.1016/j.jbiotec.2013.10.030

Cited by 30 publications

(23 citation statements)

References 22 publications

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“…The addition of a functional xylose metabolic pathway resulted in a strain that was able to co-ferment xylose and cellobiose [8]. Further efforts to increase the rate of cellobiose fermentation have included combinatorial transcriptional engineering [9], experimental evolution [10][11][12][13][14], exploration and optimization of cellodextrin transporters [15,16], and the manipulation of transcription factors [7]. Despite these engineering efforts, cellobiose fermentation by S. cerevisiae is still very limited.…”

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confidence: 99%

Metabolic engineering of the cellulolytic thermophilic fungus Myceliophthora thermophila to produce ethanol from cellobiose

Zhang

et al. 2020

Biotechnol Biofuels

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Background: Cellulosic biomass is a promising resource for bioethanol production. However, various sugars in plant biomass hydrolysates including cellodextrins, cellobiose, glucose, xylose, and arabinose, are poorly fermented by microbes. The commonly used ethanol-producing microbe Saccharomyces cerevisiae can usually only utilize glucose, although metabolically engineered strains that utilize xylose have been developed. Direct fermentation of cellobiose could avoid glucose repression during biomass fermentation, but applications of an engineered cellobiose-utilizing S. cerevisiae are still limited because of its long lag phase. Bioethanol production from biomass-derived sugars by a cellulolytic filamentous fungus would have many advantages for the biorefinery industry. Results: We selected Myceliophthora thermophila, a cellulolytic thermophilic filamentous fungus for metabolic engineering to produce ethanol from glucose and cellobiose. Ethanol production was increased by 57% from glucose but not cellobiose after introduction of ScADH1 into the wild-type (WT) strain. Further overexpression of a glucose transporter GLT-1 or the cellodextrin transport system (CDT-1/CDT-2) from N. crassa increased ethanol production by 131% from glucose or by 200% from cellobiose, respectively. Transcriptomic analysis of the engineered cellobioseutilizing strain and WT when grown on cellobiose showed that genes involved in oxidation-reduction reactions and the stress response were downregulated, whereas those involved in protein biosynthesis were upregulated in this effective ethanol production strain. Turning down the expression of pyc gene results the final engineered strain with the ethanol production was further increased by 23%, reaching up to 11.3 g/L on cellobiose. Conclusions: This is the first attempt to engineer the cellulolytic fungus M. thermophila to produce bioethanol from biomass-derived sugars such as glucose and cellobiose. The ethanol production can be improved about 4 times up to 11 grams per liter on cellobiose after a couple of genetic engineering. These results show that M. thermophila is a promising platform for bioethanol production from cellulosic materials in the future.

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confidence: 99%

Metabolic engineering of the cellulolytic thermophilic fungus Myceliophthora thermophila to produce ethanol from cellobiose

Zhang

et al. 2020

Biotechnol Biofuels

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“…Cellodextrin transporters enable the non-cellobiose fermenting S. cerevisiae and E. coli to uptake cellodextrins, facilitating the engineered microorganisms to produce biofuels (Ha et al, 2011;Vinuselvi and Lee, 2011;Bae et al, 2014;Fan et al, 2016). To further confirm the function of CtA, the fermentation of YPβG-CtA on cellobiose (20 g/L) was conducted anaerobically, and the cellobiose consumption and ethanol production were analyzed ( Figure 5).…”

Section: Ethanol Production Of Ypβg-cta On Cellobiosementioning

confidence: 99%

Identification and Characterization of a Cellodextrin Transporter in Aspergillus niger

et al. 2020

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Aspergillus niger produces a wide spectrum of extracellular polysaccharide hydrolases that hydrolyze cellulose into soluble glucose and cellodextrins. Transporters are essential for sugar uptake, yet it is not clear whether cellodextrin transporters exist in A. niger. Here, one cellulose inducible cellodextrin transporter CtA was identified in A. niger B2. It was found that CtA not only could transport cellobiose, but also cellotriose, cellotetraose, and cellopentaose. The yeast strain YPβG-CtA, expressing cellodextrin transporter CtA and an intracellular β-glucosidase, grew on cellobiose with the cell growth rate of 0.0830 ± 0.0113 h −1 under aerobic condition. Furthermore, the engineered yeast could produce 1.1 g/L ethanol anaerobically on cellobiose in 2 days. The identification of CtA provides evidence that the cellodextrin uptake is a complementary strategy of cellulose utilization in A. niger, and the CtA could be a strong transporter candidate for constructing engineered cellodextrin-utilizing microorganisms.

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“…However, S. cerevisiae contains at least eight hexose transporters (Hxt) that mediate D-glucose uptake [66], and which can also take up D-xylose albeit with lower affinity [67]. Shin et al [68] into S. cerevisiae [73][74] and Y. lipolytica [75] for which the dependence on ß-glucosidase for lignocellulose utilization was drastically reduced or eliminated.…”

Section: Industrial Uses Of Lignocellulose Hydrolysis Productsmentioning

confidence: 99%

Fungal Enzymes for Bio-Products from Sustainable and Waste Biomass

Gupta

Kubicek

Berrin

et al. 2016

Trends in Biochemical Sciences

243

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Lignocellulose, the most abundant renewable carbon source on earth, is the logical candidate to replace fossil carbon as the major biofuel raw material. Nevertheless, the technologies needed to convert lignocellulose into soluble products that can then be utilized by the chemical or fuel industries face several challenges. Enzymatic hydrolysis is of major importance, and we review the progress made in fungal enzyme technology over the past few years with major emphasis on (i) the enzymes needed for the conversion of polysaccharides (cellulose and hemicellulose) into soluble products, (ii) the potential uses of lignin degradation products, and (iii) current progress and bottlenecks for the use of the soluble lignocellulose derivatives in emerging biorefineries.

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Molecular cloning and expression of fungal cellobiose transporters and β-glucosidases conferring efficient cellobiose fermentation in Saccharomyces cerevisiae

Cited by 30 publications

References 22 publications

Metabolic engineering of the cellulolytic thermophilic fungus Myceliophthora thermophila to produce ethanol from cellobiose

Metabolic engineering of the cellulolytic thermophilic fungus Myceliophthora thermophila to produce ethanol from cellobiose

Identification and Characterization of a Cellodextrin Transporter in Aspergillus niger

Fungal Enzymes for Bio-Products from Sustainable and Waste Biomass

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