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Production of cellulosic ethanol and enzyme from waste fiber sludge using SSF, recycling of hydrolytic enzymes and yeast, and recombinant cellulase-producing Aspergillus niger
Umeå University, Faculty of Science and Technology, Department of Chemistry.
Processum Biorefinery Initiative AB, 891 22 Örnsköldsvik, Sweden.
Department of Microbiology, Stellenbosch University, Stellenbosch, 7602, South Africa.
Department of Microbiology, Stellenbosch University, Stellenbosch, 7602, South Africa.
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2014 (English)In: Journal of Industrial Microbiology & Biotechnology, ISSN 1367-5435, E-ISSN 1476-5535, Vol. 41, no 8, 1191-1200 p.Article in journal (Refereed) Published
Abstract [en]

Bioethanol and enzymes were produced from fiber sludges through sequential microbial cultivations. After a first simultaneous saccharification and fermentation (SSF) with yeast, the bioethanol concentrations of sulfate and sulfite fiber sludges were 45.6 and 64.7 g/L, respectively. The second SSF, which included fresh fiber sludges and recycled yeast and enzymes from the first SSF, resulted in ethanol concentrations of 38.3 g/L for sulfate fiber sludge and 24.4 g/L for sulfite fiber sludge. Aspergillus niger carrying the endoglucanase-encoding Cel7B gene of Trichoderma reesei was grown in the spent fiber sludge hydrolysates. The cellulase activities obtained with spent hydrolysates of sulfate and sulfite fiber sludges were 2,700 and 2,900 nkat/mL, respectively. The high cellulase activities produced by using stillage and the significant ethanol concentrations produced in the second SSF suggest that onsite enzyme production and recycling of enzyme are realistic concepts that warrant further attention.

Place, publisher, year, edition, pages
Springer Berlin/Heidelberg, 2014. Vol. 41, no 8, 1191-1200 p.
Keyword [en]
biorefinery, fiber sludge, cellulosic ethanol, enzymes, cellulase
National Category
Biocatalysis and Enzyme Technology Microbiology
URN: urn:nbn:se:umu:diva-82484DOI: 10.1007/s10295-014-1457-9ISI: 000339385400002OAI: diva2:661374

Included in thesis in manuscript form.

Available from: 2013-11-03 Created: 2013-11-03 Last updated: 2014-09-01Bibliographically approved
In thesis
1. Biorefining of lignocellulose: Detoxification of inhibitory hydrolysates and potential utilization of residual streams for production of enzymes
Open this publication in new window or tab >>Biorefining of lignocellulose: Detoxification of inhibitory hydrolysates and potential utilization of residual streams for production of enzymes
2013 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Lignocellulosic biomass is a renewable resource that can be utilized for the production of biofuels, chemicals, and bio-based materials. Biochemical conversion of lignocellulose to advanced biofuels, such as cellulosic ethanol, is generally performed through microbial fermentation of sugars generated by thermochemical pretreatment of the biomass followed by an enzymatic hydrolysis of the cellulose. The aims of the research presented in this thesis were to address problems associated with pretreatment by-products that inhibit microbial and enzymatic biocatalysts, and to investigate the potential of utilizing residual streams from pulp mills and biorefineries to produce hydrolytic enzymes.

A novel method to detoxify lignocellulosic hydrolysates to improve the fermentability was investigated in experiments with the yeast Saccharomyces cerevisiae. The method is based on treatment of lignocellulosic slurries and hydrolysates with reducing agents, such as sodium dithionite and sodium sulfite. The effects of treatment with sodium borohydride were also investigated. Treatment of a hydrolysate of Norway spruce by addition of 10 mM dithionite resulted in an increase of the balanced ethanol yield from 0.03 to 0.35 g/g. Similarly, the balanced ethanol yield of a hydrolysate of sugarcane bagasse increased from 0.06 to 0.28 g/g after treatment with 10 mM dithionite. In another study with a hydrolysate of Norway spruce, addition of 34 mM borohydride increased the balanced ethanol yield from 0.02 to 0.30 g/g, while the ethanol productivity increased from 0.05 to 0.57 g/(L×h). While treatment with sulfur oxyanions had a positive effect on microbial fermentation and enzymatic hydrolysis, treatment with borohydride resulted in an improvement only for the microbial fermentation. The chemical effects of treatments of hydrolysates with sodium dithionite, sodium sulfite, and sodium borohydride were investigated using liquid chromatography-mass spectrometry (LC-MS). Treatments with dithionite and sulfite were found to rapidly sulfonate inhibitors already at room temperature and at a pH that is compatible with enzymatic hydrolysis and microbial fermentation. Treatment with borohydride reduced inhibitory compounds, but the products were less hydrophilic than the products obtained in the reactions with the sulfur oxyanions.

The potential of on-site enzyme production using low-value residual streams, such as stillage, was investigated utilizing recombinant Aspergillus niger producing xylanase and cellulase. A xylanase activity of 8,400 nkat/ml and a cellulase activity of 2,700 nkat/ml were reached using stillages from processes based on waste fiber sludge. The fungus consumed a large part of the xylose, the acetic acid, and the oligosaccharides that were left in the stillages after fermentation with S. cerevisiae. In another study, the capability of two filamentous fungi (A. niger and Trichoderma reesei) and three yeasts (S. cerevisiae, Pichia pastoris, and Yarrowia lipolytica) to grow on inhibitory lignocellulosic media were compared. The results indicate that the two filamentous fungi had the best capability to utilize different nutrients in the media, while the S. cerevisiae strain exhibited the best tolerance against the inhibitors. Utilization of different nutrients would be especially important in enzyme production using residual streams, while tolerance against inhibitors is desirable in a consolidated bio-process in which the fermenting microorganism also contributes by producing enzymes.

Place, publisher, year, edition, pages
Umeå: Umeå universitet, 2013. 80 p.
National Category
Biocatalysis and Enzyme Technology
urn:nbn:se:umu:diva-82486 (URN)978-91-7459-759-2 (ISBN)
Public defence
2013-11-29, KBC-huset, KB3A9, Umeå universitet, Umeå, 10:15 (English)
Available from: 2013-11-08 Created: 2013-11-03 Last updated: 2013-11-08Bibliographically approved

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Cavka, AdnanJönsson, Leif J.
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