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Ammonium hydroxide detoxification of spruce acid hydrolysates.
Karlstad University, Division for Chemistry.
Karlstad University, Division for Chemistry.
Karlstad University, Division for Chemistry.
Karlstad University, Division for Chemistry.
Show others and affiliations
2005 (English)In: Applied Biochemistry and Biotechnology, ISSN 0273-2289, E-ISSN 1599-0291, Vol. 121-124, 911-22 p.Article in journal (Refereed) Published
Abstract [en]

When dilute-acid hydrolysates from spruce are fermented to produce ethanol, detoxification is required to make the hydrolysates fermentable at reasonable rates. Treatment with alkali, usually by overliming, is one of the most efficient approaches. Several nutrients, such as ammonium and phosphate, are added to the hydrolysates prior to fermentation. We investigated the use of NH4OH for simultaneous detoxification and addition of nitrogen source. Treatment with NH4OH compared favorably with Ca(OH)2, Mg(OH)2, Ba(OH)2, and NaOH to improve fermentability using Saccharomyces cerevisiae. Analysis of monosaccharides, furan aldehydes, phenols, and aliphatic acids was performed after the different treatments. The NH4OH treatments, performed at pH 10.0, resulted in a substantial decrease in the concentrations of furfural and hydroxymethylfurfural. Under the conditions studied, NH4OH treatments gave better results than Ca(OH)2 treatments. The addition of an extra nitrogen source in the form of NH4Cl at pH 5.5 did not result in any improvement in fermentability that was comparable to NH4OH treatments at alkaline conditions. The addition of CaCl2 or NH4Cl at pH 5.5 after treatment with NH4OH or Ca(OH)2 resulted in poorer fermentability, and the negative effects were attributed to salt stress. The results strongly suggest that the highly positive effects of NH4OH treatments are owing to chemical conversions rather than stimulation of the yeast cells by ammonium ions during the fermentation.

Place, publisher, year, edition, pages
2005. Vol. 121-124, 911-22 p.
National Category
Other Chemistry Topics
Research subject
Chemistry
Identifiers
URN: urn:nbn:se:kau:diva-3310DOI: 10.1385/ABAB:124:1-3:0911PubMedID: 15930570OAI: oai:DiVA.org:kau-3310DiVA: diva2:134150
Available from: 2009-01-19 Created: 2009-01-19 Last updated: 2013-10-25Bibliographically approved
In thesis
1. Ethanol from lignocellulose: Management of by-products of hydrolysis
Open this publication in new window or tab >>Ethanol from lignocellulose: Management of by-products of hydrolysis
2009 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Fuel ethanol can be produced from lignocellulosic materials, such as residues from agriculture and forestry. The polysaccharides of lignocellulose are converted to sugars by hydrolysis and the sugars can then be fermented to ethanol using microorganisms. However, during hydrolysis a wide range of by-products are also generated. By-product formation can affect ethanol yield and productivity. Management of by-products of hydrolysis is therefore important in the development of commercially viable production of cellulosic ethanol.

Detoxification of inhibitory dilute-acid lignocellulose hydrolysates by treatment with Ca(OH)2 (overliming) efficiently improves the fermentability, but is associated with drawbacks like sugar degradation and CaSO4 precipitation. In factorial designed experiments, in which pH and temperature were varied, dilute-acid spruce hydrolysates were treated with Ca(OH)2, NH4OH or NaOH. The concentrations of sugars and inhibitory compounds were measured before and after the treatments. The fermentability was examined using the yeast Saccharomyces cerevisiae and compared with reference fermentations of synthetic medium without inhibitors. The treatment conditions were evaluated by comparing the balanced ethanol yield, which takes both the degradation of sugars and the ethanol production into account. Treatment conditions resulting in excellent fermentability and minimal sugar degradation were possible to find regardless of whether Ca(OH)2, NH4OH or NaOH was used. Balanced ethanol yields higher than those of the reference fermentations were achieved for hydrolysates treated with all three types of alkali. As expected, treatment with Ca(OH)2 gave rise to precipitated CaSO4. The NH4OH treatments gave rise to a brownish precipitate but the amounts of precipitate formed were relatively small. No precipitate was observed in treatments with NaOH. The findings presented can be used to improve the effectiveness of alkali detoxification of lignocellulose hydrolysates and to minimize problems with sugar degradation and formation of precipitates.

Overexpression of different S. cerevisiae genes was investigated with the aim to engineer a biocatalyst with increased inhibitor tolerance. Overexpression of YAP1, a gene encoding a transcription factor, conveyed increased resistance to lignocellulose-derived inhibitors as well as to a dilute-acid hydrolysate of spruce.

Recombinant Aspergillus niger expressing the Hypocrea jecorina endoglucanase Cel7B was cultivated on spent lignocellulose hydrolysate (stillage). The fungus simultaneously removed inhibitors present in the stillage and produced higher amounts of endoglucanase than when it was grown in a standard medium with comparable monosaccharide content. The concept can be applied for on-site production of enzymes in a cellulose-to-ethanol process and facilitate recycling of the stillage stream.

Place, publisher, year, edition, pages
Karlstad: Karlstad University, 2009. 71 p.
Series
Karlstad University Studies, ISSN 1403-8099 ; 2009:7
Keyword
Ethanol, Lignocellulose, Detoxification, Genetic engineering, Enzyme production
National Category
Other Chemistry Topics
Research subject
Chemistry
Identifiers
urn:nbn:se:kau:diva-3314 (URN)978-91-7063-228-0 (ISBN)
Public defence
2009-02-27, Ericssonsalen, 9C 204, Karlstad Universitet, Karlstad, 13:15 (English)
Opponent
Supervisors
Available from: 2009-02-09 Created: 2009-01-19 Last updated: 2011-10-05Bibliographically approved
2. Ethanol from lignocellulose: Alkali detoxification of dilute-acid spruce hydrolysates
Open this publication in new window or tab >>Ethanol from lignocellulose: Alkali detoxification of dilute-acid spruce hydrolysates
2006 (English)Licentiate thesis, comprehensive summary (Other scientific)
Abstract [en]

Detoxification of dilute-acid lignocellulose hydrolysates by treatment with Ca(OH)2 (overliming) efficiently improves the production of fuel ethanol, but is associated with drawbacks like sugar degradation and CaSO4 precipitation. In factorial designed experiments, in which pH and temperature were varied, dilute-acid spruce hydrolysates were treated with Ca(OH)2, NH4OH or NaOH. The concentrations of sugars and inhibitory compounds were measured before and after the treatments. The fermentability was examined using the yeast Saccharomyces cerevisiae and compared with reference fermentations of synthetic medium without inhibitors. The treatment conditions were evaluated by comparing the balanced ethanol yield, which takes both the degradation of sugars and the ethanol production into account. Treatment conditions resulting in excellent fermentability and minimal sugar degradation were possible to find regardless of whether Ca(OH)2, NH4OH or NaOH was used. Balanced ethanol yields higher than those of the reference fermentations were achieved for hydrolysates treated with all three types of alkali. As expected, treatment with Ca(OH)2 gave rise to precipitated CaSO4. The NH4OH treatments gave rise to a brownish precipitate but the amounts of precipitate formed were relatively small. No precipitate was observed in treatments with NaOH. The possibility that the ammonium ions from the NH4OH treatments gave a positive effect as an extra source of nitrogen during the fermentations was excluded after experiments in which NH4Cl was added to the medium. The findings presented can be used to improve the effectiveness of alkali detoxification of lignocellulose hydrolysates and to minimize problems with sugar degradation and formation of precipitates.

Place, publisher, year, edition, pages
Karlstad: Karlstad University, 2006. 38 p.
Series
Karlstad University Studies, ISSN 1403-8099 ; 2006:30
National Category
Other Chemistry Topics
Research subject
Chemistry
Identifiers
urn:nbn:se:kau:diva-2632 (URN)91-7063-061-5 (ISBN)
Presentation
(English)
Available from: 2010-03-29 Created: 2010-03-29 Last updated: 2011-10-05Bibliographically approved

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