Dataset Information

Expression of Aspergillus niger CAZymes is determined by compositional changes in wheat straw generated by hydrothermal or ionic liquid pretreatments.

ABSTRACT: BACKGROUND:The capacity of fungi, such as Aspergillus niger, to degrade lignocellulose is harnessed in biotechnology to generate biofuels and high-value compounds from renewable feedstocks. Most feedstocks are currently pretreated to increase enzymatic digestibility: improving our understanding of the transcriptomic responses of fungi to pretreated lignocellulosic substrates could help to improve the mix of activities and reduce the production costs of commercial lignocellulose saccharifying cocktails. RESULTS:We investigated the responses of A. niger to untreated, ionic liquid and hydrothermally pretreated wheat straw over a 5-day time course using RNA-seq and targeted proteomics. The ionic liquid pretreatment altered the cellulose crystallinity while retaining more of the hemicellulosic sugars than the hydrothermal pretreatment. Ionic liquid pretreatment of straw led to a dynamic induction and repression of genes, which was correlated with the higher levels of pentose sugars saccharified from the ionic liquid-pretreated straw. Hydrothermal pretreatment of straw led to reduced levels of transcripts of genes encoding carbohydrate-active enzymes as well as the derived proteins and enzyme activities. Both pretreatments abolished the expression of a large set of genes encoding pectinolytic enzymes. These reduced levels could be explained by the removal of parts of the lignocellulose by the hydrothermal pretreatment. The time course also facilitated identification of temporally limited gene induction patterns. CONCLUSIONS:The presented transcriptomic and biochemical datasets demonstrate that pretreatments caused modifications of the lignocellulose, to both specific structural features as well as the organisation of the overall lignocellulosic structure, that determined A. niger transcript levels. The experimental setup allowed reliable detection of substrate-specific gene expression patterns as well as hitherto non-expressed genes. Our data suggest beneficial effects of using untreated and IL-pretreated straw, but not HT-pretreated straw, as feedstock for CAZyme production.

SUBMITTER: Daly P

PROVIDER: S-EPMC5294722 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

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Expression of <i>Aspergillus niger</i> CAZymes is determined by compositional changes in wheat straw generated by hydrothermal or ionic liquid pretreatments.

Daly Paul P van Munster Jolanda M JM Blythe Martin J MJ Ibbett Roger R Kokolski Matt M Gaddipati Sanyasi S Lindquist Erika E Singan Vasanth R VR Barry Kerrie W KW Lipzen Anna A Ngan Chew Yee CY Petzold Christopher J CJ Chan Leanne Jade G LJG Pullan Steven T ST Delmas Stéphane S Waldron Paul R PR Grigoriev Igor V IV Tucker Gregory A GA Simmons Blake A BA Archer David B DB

Biotechnology for biofuels 20170207

<h4>Background</h4>The capacity of fungi, such as <i>Aspergillus niger,</i> to degrade lignocellulose is harnessed in biotechnology to generate biofuels and high-value compounds from renewable feedstocks. Most feedstocks are currently pretreated to increase enzymatic digestibility: improving our understanding of the transcriptomic responses of fungi to pretreated lignocellulosic substrates could help to improve the mix of activities and reduce the production costs of commercial lignocellulose sa ...[more]

PMID: 28184248

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Project description:In this study, bagasse was pretreated with ionic liquid (IL) 1-butyl-3-methylimidazolium chloride ([Bmim]Cl) and 1% NaOH solution for initial activation of bagasse. A mixed fermentation of treated bagasse by Aspergillus niger and Candida shehatae showed the optimal conditions with the addition of C. shehatae 12 h later at a 1:1 proportion to A. niger. To further improve the ethanol production and obtain optimal fermentation conditions, a Plackett-Burman design was applied to screen the significant formulation and process variables. The optimal ethanol fermentation conditions with IL pretreated bagasse were determined using response surface methodology by Box-Behnken design. Three variables "initial pH, (NH4)2SO4, fermentation time" were regarded as significant factors in the optimization study. The resulting optimum fermentation conditions for bioethanol was identified as: initial pH of 5.89, (NH4)2SO4 concentration of 0.40 g/50 mL, and fermentation time of 3.60 days. The verification experimental ethanol concentration was 8.14 g/L, which agreed with the predicted value. An enhancement of approximately 153.58% compared with initial fermentation conditions in ethanol production was found using optimized conditions. It demonstrated that optimization methodology had a positive effect on the improvement of ethanol production. Under the optimal fermentation medium and conditions, the ethanol production with IL-pretreated bagasse and untreated bagasse was 8.14 g/L and 5.03 g/L, respectively, which exhibited 62% increase, compared to initial conditions with production of 3.21 g/L and 2.67 g/L, respectively, which displayed 20% increase. Both under optimal and original fermentation conditions, compared to the fermentation medium with untreated bagasse, all the results indicated that IL-pretreated bagasse resulted in higher ethanol production than untreated bagasse, demonstrating that IL-pretreated bagasse successfully increased the ethanol production in the mixed fermentation by A. niger and C. shehatae.

Project description:BackgroundThe use of the enzymatic hydrolysis of lignocellulose with subsequent fermentation to ethanol provides a green alternative for the production of transportation fuels. Because of its recalcitrant nature, the lignocellulosic biomass must be pretreated before enzymatic hydrolysis. However, the pretreatment often results in the formation of compounds that are inhibitory for the enzymes or fermenting organism. Although well recognized, little quantitative information on the inhibition of individual cellulase components by identified inhibitors is available.ResultsStrong cellulase inhibitors were separated from the liquid fraction of the hydrothermal pretreatment of wheat straw. HPLC and mass-spectroscopy analyses confirmed that the inhibitors were oligosaccharides (inhibitory oligosaccharides, IOS) with a degree of polymerization from 7 to 16. The IOS are composed of a mixture of xylo- (XOS) and gluco-oligosaccharides (GOS). We propose that XOS and GOS are the fragments of the xylan backbone and mixed-linkage β-glucans, respectively. The IOS were approximately 100 times stronger inhibitors for Trichoderma reesei cellobiohydrolases (CBHs) than cellobiose, which is one of the strongest inhibitors of these enzymes reported to date. Inhibition of endoglucanases (EGs) by IOS was weaker than that of CBHs. Most of the tested cellulases and hemicellulases were able to slowly degrade IOS and reduce the inhibitory power of the liquid fraction to some extent. The most efficient single enzyme component here was T. reesei EG TrCel7B. Although reduced by the enzyme treatment, the residual inhibitory power of IOS and the liquid fraction was strong enough to silence the major component of the T. reesei cellulase system, CBH TrCel7A.ConclusionsThe cellulase inhibitors described here may be responsible for the poor yields from the enzymatic conversion of the whole slurries from lignocellulose pretreatment under conditions that do not favor complete degradation of hemicellulose. Identification of the inhibitory compounds helps to design better enzyme mixtures for their degradation and to optimize the pretreatment regimes to minimize their formation.

Dataset Information

Expression of Aspergillus niger CAZymes is determined by compositional changes in wheat straw generated by hydrothermal or ionic liquid pretreatments.

Publications

Expression of <i>Aspergillus niger</i> CAZymes is determined by compositional changes in wheat straw generated by hydrothermal or ionic liquid pretreatments.

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