Nils-Olof Nilvebrant

Karlstads universitet, Karlstad, Värmland, Sweden

Are you Nils-Olof Nilvebrant?

Claim your profile

Publications (32)65.17 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: Precipitation of sparsely soluble calcium oxalate causes scaling problems in the pulp and paper industry. A potential solution is to degrade the oxalic acid using oxalate-degrading enzymes. Four novel fungal oxalate decarboxylases were evaluated in experiments with 16 pH-adjusted bleaching filtrates collected from mills producing mechanical pulp or kraft pulp. The enzymes were also tested in five of the filtrates from mechanical pulping at authentic pH and elevated temperature (55°C). The enzyme that performed best in the screening was selected for a small-scale experiment performed in a mill producing mechanical pulp. The enzyme degraded 70% of the oxalic acid in the fresh filtrate after one hour, without pH adjustment and at the prevailing process temperature (65°C). The new enzyme performed considerably better than the well-studied oxalate decarboxylase from Aspergillus niger, which only degraded 4% of the oxalic acid under the same conditions.
    Industrial Biotechnology 04/2014; 10(2):126-129. DOI:10.1089/ind.2014.0001
  • Source
    Leif J Jönsson · Björn Alriksson · Nils-Olof Nilvebrant ·
    [Show abstract] [Hide abstract]
    ABSTRACT: Bioconversion of lignocellulose by microbial fermentation is typically preceded by an acidic thermochemical pretreatment step designed to facilitate enzymatic hydrolysis of cellulose. Substances formed during the pretreatment of the lignocellulosic feedstock inhibit enzymatic hydrolysis as well as microbial fermentation steps. This review focuses on inhibitors from lignocellulosic feedstocks and how conditioning of slurries and hydrolysates can be used to alleviate inhibition problems. Novel developments in the area include chemical in-situ detoxification by using reducing agents, and methods that improve the performance of both enzymatic and microbial biocatalysts.
    Biotechnology for Biofuels 01/2013; 6(1):16. DOI:10.1186/1754-6834-6-16 · 6.04 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: The effects of ionic substances in seven industrial filtrates from kraft pulping, mechanical pulping, and sulfite pulping on the activity of oxalate oxidase from barley were investigated by pre-treatment of the filtrates with ion-exchange resins prior to enzymatic degradation of the oxalic acid in the filtrates. The pre-treatment resulted in increased oxalic acid degradation rates in all filtrates, except for one that was obtained from sulfite pulping. The possibility that lignosulfonates, which were present in the filtrate from sulfite pulping, could affect oxalate oxidase was investigated in a separate set of experiments involving four different preparations of lignosulfonates. At a lignosulfonate concentration of 50 mg/mL and a pH of 3.8, only 2–16% of the activity of oxalate oxidase remained. The results show the effects of anionic and cationic substances in bleaching filtrates on oxalate oxidase and indicate that there is an interaction between the enzyme, which has a positive net charge at pH 3.8, and the polymeric anionic lignosulfonates.
    Engineering in Life Sciences 06/2011; 11(3):245 - 252. DOI:10.1002/elsc.201000156 · 2.49 Impact Factor
  • Pierre Cassland · Anders Sjöde · Sandra Winestrand · Leif J Jönsson · Nils-Olof Nilvebrant ·
    [Show abstract] [Hide abstract]
    ABSTRACT: Increased recirculation of process water has given rise to problems with formation of calcium oxalate incrusts (scaling) in the pulp and paper industry and in forest biorefineries. The potential in using oxalate decarboxylase from Aspergillus niger for oxalic acid removal in industrial bleaching plant filtrates containing oxalic acid was examined and compared with barley oxalate oxidase. Ten different filtrates from chemical pulping were selected for the evaluation. Oxalate decarboxylase degraded oxalic acid faster than oxalate oxidase in eight of the filtrates, while oxalate oxidase performed better in one filtrate. One of the filtrates inhibited both enzymes. The potential inhibitory effect of selected compounds on the enzymatic activity was tested. Oxalate decarboxylase was more sensitive than oxalate oxidase to hydrogen peroxide. Oxalate decarboxylase was not as sensitive to chlorate and chlorite as oxalate oxidase. Up to 4 mM chlorate ions, the highest concentration tested, had no inhibitory effect on oxalate decarboxylase. Analysis of the filtrates suggests that high concentrations of chlorate present in some of the filtrates were responsible for the higher sensitivity of oxalate oxidase in these filtrates. Oxalate decarboxylase was thus a better choice than oxalate oxidase for treatment of filtrates from chlorine dioxide bleaching.
    Applied biochemistry and biotechnology 09/2009; 161(1-8):255-63. DOI:10.1007/s12010-009-8769-7 · 1.74 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: A recombinant Aspergillus niger strain expressing the Hypocrea jecorina endoglucanase Cel7B was grown on spent hydrolysates (stillage) from sugarcane bagasse and spruce wood. The spent hydrolysates served as excellent growth media for the Cel7B-producing strain, A. niger D15[egI], which displayed higher endoglucanase activities in the spent hydrolysates than in standard medium with a comparable monosaccharide content (e.g., 2,100 nkat/ml in spent bagasse hydrolysate compared to 480 nkat/ml in standard glucose-based medium). In addition, A. niger D15[egI] was also able to consume or convert other lignocellulose-derived compounds, such as acetic acid, furan aldehydes, and phenolic compounds, which are recognized as inhibitors of yeast during ethanolic fermentation. The results indicate that enzymes can be produced from the stillage stream as a high-value coproduct in second-generation bioethanol plants in a way that also facilitates recirculation of process water.
    Applied and Environmental Microbiology 03/2009; 75(8):2366-74. DOI:10.1128/AEM.02479-08 · 3.67 Impact Factor
  • Source
    Sandra Winestrand · Nils-Olof Nilvebrant · Leif J Jönsson ·
    [Show abstract] [Hide abstract]
    ABSTRACT: Oxalate oxidase catalyzes the conversion of oxalic acid and molecular oxygen to carbon dioxide and hydrogen peroxide. Oxalate-degrading enzymes are of interest for various applications including clinical analysis of the levels of oxalic acid in blood and urine and control of oxalic acid in industrial processes, such as pulp and paper manufacture. In these applications, the presence of oxyanions other than oxalate may affect the enzyme activity. The inhibitory effect of selected oxyanions on oxalate oxidase from barley was investigated. Seven out of fourteen of the compounds studied in-hibited oxalate oxidase at a concentration of 1 mM. Perchlorate, chlorate and chlorite were selected for more detailed studies. The results indicate that perchlorate, chlorate and chlorite cause mixed inhibition of oxalate oxidase and that the severity of the inhibition within the series increases with the oxidation state. The apparent K M of the enzyme was 0.28 ± 0.05 mM.
  • Anders Sjöde · Sandra Winestrand · Nils-Olof Nilvebrant · Leif J. Jönsson ·
    [Show abstract] [Hide abstract]
    ABSTRACT: Enzymatically catalyzed decomposition of oxalic acid in bleaching filtrates from the pulp and paper industry offers a possibility to enduringly prevent oxalate scaling problems by specific removal of the oxalic acid in the system rather than by attempting to avoid calcium oxalate precipitation by countermeasures aiming at improved solubility. To achieve a broad evaluation of various oxalate-degrading enzymes and to cover conditions encountered in various types of processes, 16 different bleaching filtrates were collected from pulp mills engaged in mechanical pulping of softwood, mechanical pulping of aspen, and kraft pulping of softwood. A novel oxalate-degrading enzyme provided by Novozymes was compared with commercially available oxalate oxidase from barley and oxalate decarboxylase from Aspergillus niger. The activity of the enzymes in the filtrates was investigated using kinetic analysis and multivariate data analysis. Kinetic analysis indicated that the degradation rates were governed more by inhibitors in the filtrates than by the concentration of oxalic acid. Multivariate data analysis suggested links between high concentrations of certain compounds in the filtrates and high or low enzyme activity, as exemplified by the link between high concentrations of chelators in filtrates from mechanical pulping and low activity of oxalate oxidase from barley. All three enzymes could degrade oxalic acid in all filtrates, despite the fact that very high concentrations of residual hydrogen peroxide were found in several of the filtrates.
    Enzyme and Microbial Technology 08/2008; 43(2-43):78-83. DOI:10.1016/j.enzmictec.2007.11.014 · 2.32 Impact Factor
  • Source
    Carlos Martin · Björn Alriksson · Anders Sjöde · Nils-Olof Nilvebrant · Leif J Jönsson ·
    [Show abstract] [Hide abstract]
    ABSTRACT: The potential of dilute-acid prehydrolysis as a pretreatment method for sugarcane bagasse, rice hulls, peanut shells, and cassava stalks was investigated. The prehydrolysis was performed at 122 degrees C during 20, 40, or 60 min using 2% H(2)SO(4) at a solid-to-liquid ratio of 1:10. Sugar formation increased with increasing reaction time. Xylose, glucose, arabinose, and galactose were detected in all of the prehydrolysates, whereas mannose was found only in the prehydrolysates of peanut shells and cassava stalks. The hemicelluloses of bagasse were hydrolyzed to a high-extent yielding concentrations of xylose and arabinose of 19.1 and 2.2 g/L, respectively, and a xylan conversion of more than 80%. High-glucose concentrations (26-33.5 g/L) were found in the prehydrolysates of rice hulls, probably because of hydrolysis of starch of grain remains in the hulls. Peanut shells and cassava stalks rendered low amounts of sugars on prehydrolysis, indicating that the conditions were not severe enough to hydrolyze the hemicelluloses in these materials quantitatively. All prehydrolysates were readily fermentable by Saccharomyces cerevisiae. The dilute-acid prehydrolysis resulted in a 2.7- to 3.7-fold increase of the enzymatic convertibility of bagasse, but was not efficient for improving the enzymatic hydrolysis of peanut shells, cassava stalks, or rice hulls.
    Applied biochemistry and biotechnology 05/2007; 137-140(1-12):339-52. DOI:10.1007/s12010-007-9063-1 · 1.74 Impact Factor
  • Anders Sjöde · Björn Alriksson · Leif J Jönsson · Nils-Olof Nilvebrant ·
    [Show abstract] [Hide abstract]
    ABSTRACT: Industrial production of bioethanol from fibers that are unusable for pulp production in pulp mills offers an approach to product diversification and more efficient exploitation of the raw material. In an attempt to utilize fibers flowing to the biological waste treatment, selected fiber sludges from three different pulp mills were collected, chemically analyzed, enzymatically hydrolyzed, and fermented for bioethanol production. Another aim was to produce solid residues with higher heat values than those of the original fiber sludges to gain a better fuel for combustion. The glucan content ranged between 32 and 66% of the dry matter. The lignin content varied considerably (1-25%), as did the content of wood extractives (0.2-5.8%). Hydrolysates obtained using enzymatic hydrolysis were found to be readily fermentable using Saccharomyces cerevisiae. Hydrolysis resulted in improved heat values compared with corresponding untreated fiber sludges. Oligomeric xylan fragments in the solid residue obtained after enzymatic hydrolysis were identified using matrix-assisted laser desorption ionization-time of flight and their potential as a new product of a pulp mill-based biorefinery is discussed.
    Applied biochemistry and biotechnology 05/2007; 137-140(1-12):327-37. DOI:10.1007/s12010-007-9062-2 · 1.74 Impact Factor
  • Björn Alriksson · Anders Sjöde · Nils-Olof Nilvebrant · Leif J Jönsson ·
    [Show abstract] [Hide abstract]
    ABSTRACT: Alkaline detoxification strongly improves the fermentability of dilute-acid hydrolysates in the production of bioethanol from lignocellulose with Saccharomyces cerevisiae. New experiments were performed with NH4OH and NaOH to define optimal conditions for detoxification and make a comparison with Ca(OH)2 treatment feasible. As too harsh conditions lead to sugar degradation, the detoxification treatments were evaluated through the balanced ethanol yield, which takes both the ethanol production and the loss of fermentable sugars into account. The optimization treatments were performed as factorial experiments with 3-h duration and varying pH and temperature. Optimal conditions were found roughly in an area around pH 9.0/60 degrees C for NH4OH treatment and in a narrow area stretching from pH 9.0/80 degrees C to pH 12.0/30 degrees C for NaOH treatment. By optimizing treatment with NH4OH, NaOH, and Ca(OH)2, it was possible to find conditions that resulted in a fermentability that was equal or better than that of a reference fermentation of a synthetic sugar solution without inhibitors, regardless of the type of alkali used. The considerable difference in the amount of precipitate generated after treatment with different types of alkali appears critical for industrial implementation.
    Applied Biochemistry and Biotechnology 02/2006; 129-132(1-3):599-611. DOI:10.1007/978-1-59745-268-7_48 · 1.74 Impact Factor
  • Source
    Feng HONG · Anders SJÖDE · Nils-Olof NILVEBRANT · Leif J. JÖNSSON ·
    [Show abstract] [Hide abstract]
    ABSTRACT: Bleaching with oxygen-containing agents and recirculation of process streams in the pulp and paper industry has increased the accumulation of oxalic acid and danger for precipitation of calcium oxalate encrusts, scaling. Analysis and control of oxalic acid in bleaching filtrates is therefore becoming increasingly important in the pulp and paper industry. Chromatographic methods, such as IC and HPLC, are generally more time-consuming but are valuable as standard methods for determination of oxalic acid. However, the instrumentation needed is expensive and stationary. In this study, an enzymatic method based on oxalate oxidase and peroxidase was developed to determine oxalic acid in authentic bleaching filtrates using a spectrophotometer. The results showed that bleaching filtrates contain some compounds interfering with the enzymatic method. Pretreatment of the samples with activated charcoal was a successful approach for decreasing problems with interference. By using dilution followed by charcoal treatment, the results obtained from five bleaching filtrates with the colorimetric method correlated very well with those obtained using IC. This study offers a selective, fast and mobile analysis method to determine oxalic acid in bleaching filtrates from the pulp and paper industry. The convenient enzyme-based method improves the possibilities for control of critical oxalic acid concentrations in closed-loop bleaching streams.
  • Source
    Feng HONG · Anders REIMANN · Leif J. JÖNSSON · Nils Olof NILVEBRANT ·
    [Show abstract] [Hide abstract]
    ABSTRACT: Analysis and control of oxalic acid in bleaching filtrates has recently gained considerable attention in the pulp and paper industry due to problems with the formation of calcium oxalate, scaling. Chromatographic methods, such as HPLC and ion chromatography (IC), are generally valuable as standard methods for determination of oxalic acid. In this study, an HPLC system equipped with an Aminex HPX-87H column was applied to determine oxalic acid in authentic bleaching filtrates. An established IC method based on separation with an anion-exchange column was used as reference method. The results showed that bleaching filtrates contain compounds interfering with the HPLC method. A strategy, dilution of the samples followed by treatment with activated carbon, was needed to obtain similar oxalic acid concentrations as provided by the reference method. The correlation (R=0.994) between the HPLC method and the IC method is described by the equation y=1.294 7x. The method improves the possibilities for control of critical oxalic acid concentrations in closed-loop bleaching streams. 近年来草酸钙(草酸垢)的形成在制浆造纸工业中造成诸多问题,因此如何分析及控制漂白废液中的草酸浓度就显得尤为重要。高效液相色谱和离子交换色谱等色谱法一般可以作为测定草酸的标准方法。本研究以一个基于阴离子交换柱的离子交换色谱法作为对照方法,利用一套配备了Aminex HPX-87H液相色谱柱的高效液相色谱系统测定漂白废液中的草酸浓度。研究结果显示漂白废液中含有一些干扰高效液相色谱法测定的化合物。通过采用稀释样品后再经活性炭吸附的处理方法,可以得到较为满意的结果。分析发现高效液相色谱法与离子交换色谱(对照法)之间的相关性较好,相关系数为0.994。该方法的建立将有利于监控制浆造纸企业中闭路循环漂液中形成草酸钙时的临界草酸浓度。
  • [Show abstract] [Hide abstract]
    ABSTRACT: Bioethanol can be produced from wood via acid hydrolysis, but detoxification is needed to achieve good fermentability. Overliming was investigated in a factorial designed experiment, in which pH and temperature were varied. Degradation of inhibitory furan aldehydes was more extensive compared to monosaccharides. Too harsh conditions led to massive degradation of sugars and formation of inhibiting acids and phenols. The ethanol productivity and yield after optimal overliming reached levels exceeding reference fermentations of pure glucose. A novel metric, the balanced ethanol yield, which takes both ethanol production and losses of fermentable sugars into account, was introduced and showed the optimal conditions within the investigated range. The findings allow process technical and economical considerations to govern the choice of conditions for overliming.
    Applied Biochemistry and Biotechnology 02/2005; 121-124(1):1031-44. DOI:10.1385/ABAB:124:1-3:1031 · 1.74 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: 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.
    Applied Biochemistry and Biotechnology 02/2005; 121-124(1-3):911-22. DOI:10.1007/978-1-59259-991-2_78 · 1.74 Impact Factor
  • A. Sjöde · L.J. Jönsson · N.-O. Nilvebrant ·
    [Show abstract] [Hide abstract]
    ABSTRACT: Formation of oxalic acid during bleaching of pulp generates problems with calcium oxalate scaling when process water is recirculated. It is therefore important to identify conditions that minimise the formation of oxalic acid. Peracetic acid formed during a P-stage can be important for the oxalic acid formation. Thus the influence of alkali dosage, hydrogen peroxide and peracetic acid on the generation of oxalic, acetic and formic acid during peroxide bleaching of grinded softwood TMP was evaluated. Corresponding studies were also done with model substrates for several pulp components -i.e., cellulose, spruce lignin, spruce glucomannan and xylan with low acetyl content. In the pulp experiments, hydrogen peroxide in combination with high alkalinity gave the highest concentrations of oxalic acid, approx. 1 g / kg pulp. Peracetic acid contributed less to the formation of oxalic acid. Among the model substrates, xylan generated the highest amounts of oxalic acid. Peracetic acid was important for the formation of oxalic acid from cellulose, whereas hydrogen peroxide was more important in the lignin samples. In conclusion, the results suggested that hemicelluloses, particularly xylan, serve as principal precursors to oxalic acid in P-stage bleaching.
    Appita Annual Conference 01/2005; 2:303-309.
  • Pierre Cassland · Simona Larsson · Nils-Olof Nilvebrant · Leif J Jönsson ·
    [Show abstract] [Hide abstract]
    ABSTRACT: Oxalate oxidase catalyses the degradation of oxalic acid to carbon dioxide and hydrogen peroxide and is of commercial importance for clinical analyses of oxalate in biological samples. Novel potential applications for oxalate oxidase include the prevention of the formation of calcium oxalate incrusts in pulp and paper manufacture and rapid determination of oxalic acid in process waters. The potential in using oxalate-degrading enzymes in industrial processes increases the interest in finding systems for heterologous expression. Oxalate oxidase from barley is a secreted multimeric glycosylated manganese-containing enzyme with several disulfide bridges, which have been found to be essential for the catalytic activity. Attempts to achieve expression of active heterologous oxalate oxidase in bacteria have up to now met little success. In this study, one oxalate-oxidase-encoding cDNA from barley and two from wheat were cloned and tested with regard to expression in Escherichia coli. The results suggest that the selection of a novel commercially available E. coli host strain, which has the ability to form disulfide bridges in heterologous proteins expressed in its cytoplasm, was important for successful expression. Although a considerable part of the heterologous protein was produced in an insoluble and inactive form, this strain, E. coli Origami B(DE3), in addition yielded soluble and active barley and wheat oxalate oxidase. One of the wheat cDNAs, Ta(M)OXO1, gave three-fold higher activity than the barley cDNA, Hv(H)OXO1, while the other wheat cDNA, Ta(M)OXO2, gave no detectable activity. This indicates that the choice of cDNA was also critical despite the high identity between the cDNAs and the encoded polypeptides (88-89% on the nucleotide level and 88-92% on the amino-acid level). Gel filtration of cell extracts containing heterologous barley and wheat oxalate oxidase resulted in an increase in the activity. This indicates that low molecular weight inhibitory compounds were present in the E. coli lysates but could be removed by the introduction of a purification step.
    Journal of Biotechnology 05/2004; 109(1-2):53-62. DOI:10.1016/j.jbiotec.2003.10.026 · 2.87 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Six anion-exchange resins with different properties were compared with respect to detoxification of a dilute-acid hydrolysate of spruce prior to ethanolic fermentation with Saccharomyces cerevisiae. The six resins encompassed strong and weak functional groups as well as styrene-, phenol-, and acrylic-based matrices. In an analytical experimental series, fractions from columns packed with the different resins were analyzed regarding pH, glucose, furfural, hydroxymethylfurfural, phenolic compounds, levulinic acid, acetic acid, formic acid, and sulfate. An initial adsorption of glucose occurred in the strong alkaline environment and led to glucose accumulation at a later stage. Acetic and levulinic acid passed through the column before formic acid, whereas sulfate had the strongest affinity. In a preparative experimental series, one fraction from each of six columns packed with the different resins was collected for assay of the fermentability and analysis of glucose, mannose, and fermentation inhibitors. The fractions collected from strong anion-exchange resins with styrene-based matrices displayed the best fermentability: a sevenfold enhancement of ethanol productivity compared with untreated hydrolysate. Fractions from a strong anion exchanger with acrylic-based matrix and a weak exchanger with phenol-based resin displayed an intermediate improvement in fermentability, a four- to fivefold increase in ethanol productivity. The fractions from two weak exchangers with styrene- and acrylic-based matrices displayed a twofold increase in ethanol productivity. Phenolic compounds were more efficiently removed by resins with styrene- and phenol-based matrices than by resins with acrylic-based matrices.
    Applied Biochemistry and Biotechnology 02/2004; 113-116(1-3):525-38. DOI:10.1385/ABAB:114:1-3:525 · 1.74 Impact Factor
  • Source
    Feng Hong · Nils Olof Nilvebrant · Leif J Jönsson ·
    [Show abstract] [Hide abstract]
    ABSTRACT: A new method for rapid determination of oxalic acid was developed using oxalate oxidase and a biosensor based on SIRE (sensors based on injection of the recognition element) technology. The method was selective, simple, fast, and cheap compared with other present detection systems for oxalate. The total analysis time for each assay was 2-9 min. A linear range was observed between 0 and 5 mM when the reaction conditions were 30 degrees C and 60 s. The linear range and upper limit for concentration determination could be increased to 25 mM by shortening the reaction time. The lower limit of detection in standard solutions, 20 microM, could be achieved by means of modification of the reaction conditions, namely increasing the temperature and the reaction time. The biosensor method was compared with a conventional commercially available colorimetric method with respect to the determination of oxalic acid in urine samples. The urine oxalic acid concentrations determined with the biosensor method correlated well (R=0.952) with the colorimetric method.
    Biosensors & Bioelectronics 09/2003; 18(9):1173-81. DOI:10.1016/S0956-5663(02)00250-6 · 6.41 Impact Factor
  • Source
    Nils-Olof Nilvebrant · Per Persson · Anders Reimann · Filipe De Sousa · Lo Gorton · Leif J Jönsson ·
    [Show abstract] [Hide abstract]
    ABSTRACT: In addition to fermentable sugars, dilute-acid hydrolysates of lignocellulose contain compounds that inhibit fermenting microorganisms, such as Saccharomyces cerevisiae. Previous results show that phenolic compounds and furan aldehydes, and to some extent aliphatic acids, act as inhibitors during fermentation of dilute-acid hydrolysates of spruce. Treatment of lignocellulose hydrolysates with alkali, usually in the form of overliming to pH 10.0, has been frequently employed as a detoxification method to improve fermentability. A spruce dilute-acid hydrolysate was treated with NaOH in a factorial design experiment, in which the pH was varied between 9.0 and 12.0, the temperature between 5 and 80 degrees C, and the time between 1 and 7 h. Already at pH 9.0, >25% of the glucose was lost when the hydrolysate was treated at 80 degrees C for 1 h. Among the monosaccharides, xylose was degraded faster under alkaline conditions than the hexoses (glucose, mannose, and galactose), which, in turn, were degraded faster than arabinose. The results suggest that alkali treatment of hydrolysates can be performed at temperatures below 30 degrees C at any pH between 9.0 and 12.0 without problems with sugar degradation or formation of inhibiting aliphatic acids. Treatment with Ca(OH)2 instead of NaOH resulted in more substantial degradation of sugars. Under the harsher conditions of the factorial design experiment, the concentrations of furfural and 5-hydroxymethylfurfural decreased while the total phenolic content increased. The latter phenomenon was tentatively attributed to fragmentation of soluble aromatic oligomers in the hydrolysate. Separate phenolic compounds were affected in different ways by the alkaline conditions with some compounds showing an increase in concentration while others decreased. In conclusion, the conditions used for detoxification with alkali should be carefully controlled to optimize the positive effects and minimize the degradation of fermentable sugars.
    Applied Biochemistry and Biotechnology 02/2003; 105 -108(1):615-28. DOI:10.1385/ABAB:107:1-3:615 · 1.74 Impact Factor
  • Source
    Per Persson · Jessica Andersson · Lo Gorton · Simona Larsson · Nils-Olof Nilvebrant · Leif J Jönsson ·
    [Show abstract] [Hide abstract]
    ABSTRACT: Treatment with alkali, particularly overliming, has been widely used as a method for the detoxification of lignocellulose hydrolysates prior to ethanolic fermentation. However, the mechanisms behind the detoxification effect and the influence of the choice of cation have not been well understood. In this study, a dilute acid hydrolysate of spruce and an inhibitor cocktail consisting of six known inhibitors were used to investigate different alkali detoxification methods. The various treatments included the addition of calcium hydroxide, sodium hydroxide, potassium hydroxide, and ammonia to pH 10.0 and subsequent adjustment of the pH to 5.5 with either sulfuric or hydrochloric acid as well as treatment with the corresponding amounts of calcium, sodium, and potassium as sulfate or chloride salts at pH 5.5. An RP-HPLC method was developed for the separation of 18 different inhibitors in the hydrolysate, including furaldehydes and phenolics. Detection and quantification were carried out by means of UV, DAD, and ESI-MS in negative mode. Treatment of the spruce hydrolysate with alkali resulted in up to approximately 40% decrease in the concentration of furaldehydes. The effects on the aromatic compounds were complex. Furthermore, SFE was performed on the precipitate formed during alkali treatment to evaluate the inhibitor content of the precipitate, and the following RP-HPLC analysis implied that potential inhibitors were removed mainly through conversion rather than through filtration of precipitate. Parallel experiments in which sulfuric acid or hydrochloric acid was used for acidification to pH 5.5 after alkali treatment indicated that the choice of anion did not affect the removal of inhibitors. Detoxification with calcium hydroxide and ammonia resulted in better fermentability using Saccharomyces cerevisiae than detoxification with sodium hydroxide. The results from the experiments with the inhibitor cocktail indicated that the positive effects of alkali treatment are difficult to explain by removal of the inhibitors only and that possible stimulatory effects on the fermenting organism warrant further attention.
    Journal of Agricultural and Food Chemistry 10/2002; 50(19):5318-25. DOI:10.1021/jf025565o · 2.91 Impact Factor

Publication Stats

2k Citations
65.17 Total Impact Points


  • 2002-2014
    • Karlstads universitet
      • Department of Engineering and Chemical Sciences
      Karlstad, Värmland, Sweden
  • 2013
    • Borregaard
      Sarpsborg, Østfold, Norway
  • 2009
    • Umeå University
      • Department of Chemistry
      Umeå, Västerbotten, Sweden
  • 2001-2002
    • Lund University
      • Division of Applied Microbiology
      Lund, Skane, Sweden