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In partnership work, boundary spanning is crucial for breaking silo thinking and sustaining focus on inter-organizational targets. However, the literature is agency biased and focuses primarily on the role of individual boundary spanners. We present a complementary structural perspective emphasizing resource allocation, formation of coupling competence and institutionalized support from management. The relationship between structures, boundary spanning and partnership performance is examined using a case study of collaboration across Danish adult education, employment and business support services intended to boost training in private firms. Our findings generally affirm the theoretical propositions and illustrate two viable pathways for organizing partnerships.

Sugars are a sustainable platform for producing chemicals and polymers, while cellulose nanofibers (CNF) are sustainable materials with high specific mechanical properties that have an important role to play in many applications. This study highlights the feasibility of producing sugars and CNF using high dry matter (DM) content of 20 %. A commercial enzyme cocktail at low dosage, 5 mg EP/g DM, and a high 15 mg EP/g DM, were used to hydrolyze the pulp cellulose to produce sugars and CNF. HPLC was used to evaluate the cellulose conversion rate and amount of sugar realized. The CNF were studied using TEM, the diameter of the CNF were measured and estimate the crystallinity. The cellulose conversion plateaued at about 48 h for both the low and high dosage, indicating inhibition due to higher sugar concentration and/or increased recalcitrance of the remaining CNF. The CNF obtained at low enzyme indicated the fibrils were insufficiently separated. At the high dosage, less clumps were observed and with smaller diameter than with the low enzyme dosage. The high dosage also produced CNF with significantly higher crystallinity index, confirming that amorphous cellulose portions readily underwent enzymatic hydrolysis and left more recalcitrant CNF.

De planlagte og indledte trepartsforhandlinger i 2012 blev hurtigt afl yst igen. Det blev et forlis. I offentligheden er en pressemeddelelse fra Dansk Metal med erklæret modstand mod at skulle afgive hellig- og feriedage som betaling for forhandlinger gjort til grunden til sammenbruddet. Det har både i offentligheden og i den politiske historieskrivning sat sig som hovedbegrundelsen. Artiklen viser ikke kun gennem rekonstruktion af hændelsesforløbet, at det er en forenklet antagelse, men søger at stykke elementer sammen til en samlet forklaring på dette historisk enestående sammenbrud i trepartsforhandlinger. Forklaringer søges etableret gennem analyser af forhandlingsforløb, institutionelt set-up og aktørstrategier, hvor især de sidste rummer nøgle til at forstå, hvorfor sammenbruddet blev en realitet. Samspillet mellem faglige, politiske og administrative strategier har været udslagsgivende herfor. Undersøgelsen baserer sig på et unikt empirisk undersøgelsesmateriale, der ud over omfattende dokumentstudier omfatter en lang række kvalitative enkeltinterviews i perioden 2013-2017 med involverede ministre, embedsmænd, organisationsledere, cheføkonomer og forhandlere. Interpretationsfolien er en viderebearbejdning af korporatisme-, regulerings- og policy procesteori, og der søges uddraget en bredere erfaringslære af det mislykkede og fejlprogrammerede politisk bytte mellem en regering og arbejdsmarkedets parter i 2012

Reduction of the amount of nitrogen (N) remaining in mature wheat straw is an essential challenge in order to boost grain protein and N use efficiency. However, very limited information is available on the composition of the residual N pools and how they are affected by N fertilization. Winter wheat was grown under field conditions at five different N levels ranging from 60 to 280 kg N ha−1. The fraction of straw N present in amino acids, chlorophyll, DNA, nitrate and lignin-associated compounds were analyzed separately in leaf blades, sheaths and stems. Total straw N concentration increased from 0.29 to 0.69% with increasing level of N application. The largest N pool consisted of amino acids in hydrolysable proteins, which comprised 50 to 70% of the residual N. Lignin-associated, non-hydrolysable N compounds constituted 13 to 16% of the total N, while non-protein N pools (mainly chlorophyll breakdown products and DNA) amounted to 5–14%. Protein-derived amino acids constituted the most abundant N pool in mature wheat straw. Despite a 2.5-fold increase in total straw N content, the relative proportion of the different residual N pools to the total N content in the straw did not change with N fertilization.

Poul Ekelund arbejdede som maler med en symbiotisk forståelse af kultur og natur og med en særskilt måde at disponere billeder og farvestof på, som gør ham aktuel i dag.

Perennial grasses can provide very high biomass yields and a better environmental profile compared to most annual cereal crops. The grass biomass can be processed into a protein-rich fraction for monogastric feed and a fibrous pulp fraction for ruminant feed or energy production. We studied the effect of fertilization of tall fescue grown on two sandy sites in Denmark. Dry matter (DM) and crude protein (CP) yield of first cut was measured at four nitrogen (N) fertilization levels (0, 70, 140 or 210 kg N ha−1). Samples were processed in a screw press, achieving a pulp fraction and a juice fraction from which proteins were precipitated into a protein concentrate fraction. CP allocation between the fractions of protein concentrate and pulp was analysed as well as the amino acid (AA) profile. CP in the brown juice fraction was not analysed. CP concentration and CP yield of unprocessed tall fescue increased linearly with N fertilization, attaining concentrations up to 126 g kg−1 DM and yields up to 0.76-0.86 Mg ha−1. N fertilization did not affect CP allocation between concentrate and pulp in a consistent way. As a mean of all treatments and both sites, 17% of CP was recovered in the concentrate and 74% remained in the pulp. CP concentration increased linearly with fertilization in both concentrate and pulp with coefficients of 0.50 and 0.21 g kg−1 DM kg−1 N ha−1, i.e. with a higher increase in the concentrate fraction. The AA profile only differed slightly between the two fractions and between the fertilization levels, however, the total sulphur to N ratio in both fractions decreased with increased fertilization, and a similar trend was observed for the relative content of the sulphur-containing amino acids. The AA profile deviated considerably from soybean meal, particularly with higher leucine and lower glutamic acid content. In conclusion, fertilization can greatly increase CP concentration and CP yield. However, the separation process needs improvements to increase the protein in the concentrate fraction.

Sammenligning af aktiveringspolitikker i Tyskland, Danmark og UK, hvor der sker paradigmeskift på forskellige tidspunkter og med forskellige institutionelle nedslag

This is the first study investigating the effects of lignosulfonate (LS) properties and reaction conditions on the precipitation of proteins from green biomass juice for producing monogastric animal feed. Both the improvement in the protein precipitation yield and the resulting impact on the quality of the protein pellets were investigated with new methods and approaches. High molecular weight (MW) LS gave the highest precipitation yield, 21–33% higher than traditional heat and isoelectric methods. Electrostatic interaction between LS and soluble proteins was observed to be higher in high MW LS than others, and likely improved the precipitation. A response surface design experiment showed that pH was more important than LS concentration for precipitation. The optimum conditions predicted by the model improved precipitation by up to 61% compared to traditional methods. Using high MW LS at near optimum conditions resulted in more LS being retained in the pellet, with a final LS content of up to 60 g/kg dry matter (DM). The improved precipitation yield came at the expense of lower crude protein (CP) content of the pellet. Despite reduced pellet protein purity, the total CP recovery from the juice was improved by up to 800 g/kg DM. The in vitro protein digestibility (IVPD) and amino acid composition were not significantly affected by the presence of LS. Using high MW LS at optimum reaction conditions can significantly improve protein precipitation yield from green biomass juice without compromising digestibility. However, finding the compromise between yield and protein purity is necessary to improve feasibility.

Agricultural residues, such as straw, offer an opportunity to produce biofuels and chemicals in biorefineries without compromising food production. The ideal “dual‐purpose cultivar” would have high yield of grain and straw. In addition, the straw should be easy to process in a biorefinery: It should have good degradability, high concentration of carbohydrates, and low concentration of ash. Nitrogen (N) is an essential nutrient important for plant growth, crop yield and grain quality. However, N production and application comes with a high cost and high environmental footprint. The N application should consequently be based on an economical optimum. Limited knowledge exists on how N application affects the potential of straw for biorefining, for example, straw yield and quality. This study, conducted over three cropping seasons, investigated the effect of N supply on the biorefining potential and included 14 wheat cultivars and one triticale cultivar. The N supply directly affected the yield of straw and grain. In addition, the protein concentration in grain and straw increased, but the composition of the straw with respect to carbohydrates and lignin was largely unaffected by N supply. The only significant change was a lower silicon concentration at increasing N application rate, which could be beneficial for lignin valorization in biorefineries. Likely due to the negligible changes in cell wall composition, the effect of N application rate on straw degradability was not significant. N application should therefore primarily be optimized with respect to grain quality and overall yield of grain and straw. Differences between cultivars were also minor with respect to their performance in a biorefinery process. From a breeding and agronomic perspective, focus should therefore be put on maximizing the biomass output from the field, that is, selecting the cultivar with highest grain and straw yield and optimizing the application of fertilizer to get optimum N use efficiency.

The purpose of the present study was to assess the efficiency of enzyme-assisted nanofiltration for separation of xylose from glucose present in genuine biorefinery liquors obtained from hydrothermal pretreatment of wheat straw, corn stover and Miscanthus stalks. Glucose oxidase and catalase were used to convert the glucose contained in the liquors into gluconic acid, so xylose could be more easily recovered in the subsequent nanofiltration. Subjecting the biomass liquors to dilute acid treatment and centrifugation before the enzymatic reaction and filtration led to maximum biocatalytic performance of the membrane bioreactor (neglectable fouling and no enzyme activity loss) during five consecutive reaction-filtration cycles. The best separation factor of gluconic acid over xylose in the subsequent nanofiltration was 2.7, 2.5 and 2.2 for wheat straw, corn stover and Miscanthus stalks, respectively. All represented a significant improvement compared to the benchmark separation of xylose and glucose, in which case the separation factor was only 1.4. However, the higher ionic strength of the biomass liquors compared to the pure model solution probably led to a less negative zeta potential of the nanofiltration membrane, which significantly reduced the xylose purification performance as compared to the model system, for which the separation factor was 34.

Prekarisering er blevet en fællesbetegnelse for forskellige nye former for usikkerheder og utrygheder, som lønmodtagere bliver sat i på arbejdsmarkederne. Der synes imidlertid at være flere slags diagnoser indbygget i de nyere teoretiseringsforsøg, og der argumenteres i artiklen for, at det er påkrævet at bryde prekarisering op i specifikke årsag-virkningssammenhænge, som kan underkastes historisk-empirisk analyse. Social dumping og indsats herimod er en sådan sammenhæng, der dog ikke er ny, men som har fået nye udtryk og stærkere udbredelse siden Østudvidelsen i 2004. En evaluering af Københavns Kommunes indsats mod social dumping danner den empiriske basis for refleksioner over, hvor væsentligt det er, at også kommuner bliver fast institutionel del af indsatsen mod social dumping, og hvordan kommunerne udvikler deres konkrete praksisser heroverfor. Evalueringen har været med til at resultere i, at Københavns Kommune omlægger deres indsats fra at have været eksternt organiseret til at blive en intern og styrket organisering med andre operationsmåder og nye tiltag indbygget for at finde, fange og forhindre social dumping.

In order to maximize enzymatic xylan depolymerization while simultaneously purifying the resulting monosaccharide (xylose), different ultrafiltration (UF) membrane reactor configurations were evaluated. Initial results showed that the two hydrolytic enzymes required for complete depolymerization of xylan, endo-1,4-β-xylanase and β-xylosidase, promoted different types of fouling, which had a direct impact on the extent of xylan hydrolysis achieved during reaction. Endo-1,4-β-xylanase generated DP 1–6 xylo-oligomers. These products contributed to partial pore blocking of the 1 kDa polysulfone membrane and caused irreversible flux loss (∼20%). The presence of β-xylosidase could not prevent deposition of xylan and xylooligomers on the UF membrane surface. Mulder’s modelling of the filtration parameters affirmed that this xylan and xylooligosaccharide deposition formed a cake layer on the membrane which hindered enzymatic attack in addition to fouling. Reaction with both enzymes followed by UF was found to be the optimal configuration, providing at least 40% higher xylan hydrolysis than the cascade configuration (involving sequential reaction with each of the enzymes separately) and the simultaneous reaction-filtration with both enzymes, respectively. This study thus confirmed that the reactor configuration has a crucial impact on the performance of both the reaction and the separation process of xylose during enzymatic xylan degradation, and that the type of fouling mechanism varies in response to the type of enzyme treatment.

Two model solutions and a real stream from the hydrothermal pretreatment of wheat straw were subjected to nanofiltration, and permeate flux, retention and resistance to fouling were evaluated. Three commercial NF membranes were tested, and a pressure of 4 bars (range: 1–20 bars) and a temperature of 20oC (range: 20–50oC) were found to provide the best results in terms of retention. A subsequent nanodiafiltration consisting of five cycles enabled one to recover 90% of the monosaccharides (purity >99%). This result showed that diananofiltration could be a promising strategy for the recovery of high-purity streams of monosaccharides from pretreatment liquids.

Activation has no real answer to the new challenges in the labour market after covid-19 and new efforts are to be foreseen.

Key message: This study demonstrates that an active breeding nursery with rotation can be used to identify marker–trait associations for biomass yield and quality parameters that are important for biorefinery purposes. Abstract: Wheat straw is a valuable feedstock for bioethanol production, but due to the recalcitrant nature of lignocellulose, its efficient use in biorefineries is limited by its low digestibility and difficult conversion of structural carbohydrates into free sugars. A genome-wide association study (GWAS) was conducted to search for significant SNP markers that could be used in a breeding programme to improve the value of wheat straw in a biorefinery setting. As part of a 3-year breeding programme (2013–2016), 190 winter wheat lines were phenotyped for traits that affect the yield and quality of the harvested biomass. These traits included straw yield, plant height, lodging at three growth stages and Septoria tritici blotch (STB) susceptibility. Release of glucose, xylose and arabinose was determined after hydrothermal pretreatment and enzymatic hydrolysis of the straw. The lines were genotyped using 15 K SNP markers and 5552 SNP markers could be used after filtering. Heritability for all traits ranged from 0.02 to 0.74. GWASs were conducted using CMLM, SUPER and FarmCPU algorithms, to analyse which algorithm could detect the highest number of marker–trait associations (MTAs). Comparable tendencies were obtained from CMLM and FarmCPU, but FarmCPU produced the most significant results. MTAs were obtained for lodging, harvest index, plant height, STB, glucose, xylose and arabinose at a significance level of p < 9.01 × 10−6. MTAs in chromosome 6A were observed for glucose, xylose and arabinose, and could be of importance for increasing sugar release for bioethanol production.

Tryptophan is a key component in many biological processes and an essential amino acid in food and feed materials. Analysis of the tryptophan content in proteins or protein-containing matrices has always been a challenge. We show here that the preparation of samples prior to tryptophan analysis can be significantly simplified, and the time consumption reduced, by using ascorbic acid as antioxidant to eliminate the problem of tryptophan degradation during alkaline hydrolysis. Combined with separation by HPLC and detection by Single Quadrupole Mass Spectrometry, this allows the analytical run time to be reduced to 10 min. The alkaline hydrolysate obtained in the method presented here may be combined with the oxidized hydrolysate obtained when sulfur-containing amino acids are to be measured, thus essentially providing two analyses for the time of one.

Utilizing saline soil and water resources are essential in meeting future demands for food, feed, and biofuels in a world of increasing freshwater scarcity, by use of halophytes (salt-loving plants). The succulent halophyte Salicornia bigelovii Tor. was cultivated as a green biomass crop irrigated with saline aquaculture effluents, and bio-fractionation was tested as a way to improve the value of the crop. Bio-fractionation proved to be an effective way of extracting ash and nitrogen into the green juice fraction, from which a protein concentrate was produced with 15 % crude protein and 33 % ash. The fiber fraction was not found useful as fodder due to a relatively low crude protein content of 4.1 % and 10–12 % remaining ash. However, the high digestibility of the structural carbohydrates in the fiber demonstrated its potential as a feedstock for biorefining with estimated bioethanol yields around 2500 L/ha.

Membrane separation of xylose and glucose can be accomplished via oxidation of glucose to gluconic acid by enzymatic glucose oxidase catalysis. Oxygen for this reaction can be supplied via decomposition of hydrogen peroxide by enzymatic catalase catalysis. In order to maximize the biocatalytic productivity of glucose oxidase and catalase (gluconic acid yield per total amount of enzyme) the following system set-ups were compared: immobilization of glucose oxidase alone; co-immobilization of glucose oxidase and catalase; glucose oxidase and catalase free in the membrane bioreactor. Fouling-induced enzyme immobilization in the porous support of an ultrafiltration membrane was used as strategy for entrapment of glucose oxidase and catalase. The biocatalytic productivity of the membrane reactor was found to be highly related to the oxygen availability, which in turn depended on the reactor configuration, hydrogen peroxide concentration and catalase origin. When glucose oxidase and catalase (from Aspergillus niger ) were free in the membrane bioreactor a total biocatalytic productivity of 122 mg gluconic acid/mg enzyme was obtained after five consecutive reaction cycles. The free enzymes showed superior performance compared to the immobilized systems as a result of limited substrate and product diffusion in the latter case.

Commercial production of ethanol from lignocellulosic biomass is becoming a reality, but the next step is to diversify the process and produce chemicals and materials. These lignocellulosic biorefineries will in many cases rely on hydrolysis of biomass carbohydrates into monosaccharides – the sugar platform. Cellulases are the most important enzymes required in this process, but the complex nature of lignocellulose requires several other enzymes (hemicellulases and auxiliary enzymes) for efficient hydrolysis. Enzyme recycling increases the catalytic productivity of the enzymes by reusing them for several batches of hydrolysis, and thereby reduces the overall cost associated with the hydrolysis. Research on this subject has been ongoing for many years and several promising technologies and methods have been developed and demonstrated. But only in a very few cases have these technologies been upscaled and tested in industrial settings, mainly because of many difficulties with recycling of enzymes from the complex lignocellulose hydrolyzate at industrially relevant conditions, i.e., high solids loadings. The challenges are associated with the large number of different enzymes required for efficient hydrolysis, enzyme stability, and the detrimental interaction between enzyme and lignin. This review provides a comprehensive overview of the various methods for enzyme recovery and recycling, for example recycling of free enzymes, readsorption to fresh material, recycling of solids, membrane filtration, and immobilization. Lignin is a major obstacle for successful hydrolysis and enzyme recycling. A thorough understanding of this subject and possibilities to minimize adsorption or methods to desorb the enzymes are important in order to develop the technology. © 2016 Society of Chemical Industry and John Wiley & Sons, Ltd

The activity and substrate degradation pattern of a novel Aspergillus nidulans GH26 endo-β-mannanase (AnMan26A) was investigated using two galactomannan substrates with varying amounts of galactopyranosyl residues. The AnMan26A was characterized in parallel with the GH26 endomannanase from Podospora anserina (PaMan26A) and three GH5 endomannanases from A. nidulans and Trichoderma reesei (AnMan5A, AnMan5C and TrMan5A). The initial rates and the maximal degree of enzymatically catalyzed conversion of locust bean gum and guar gum galactomannans were determined. The hydrolysis product profile at maximal degree of conversion was determined using DNA sequencer-Assisted Saccharide analysis in High throughput (DASH). This is the first reported use of this method for analyzing galactomannooligosaccharides. AnMan26A and PaMan26A were found to have a novel substrate degradation pattern on the two galactomannan substrates. On the highly substituted guar gum AnMan26A and PaMan26A reached 35-40% as their maximal degree of conversion whereas the three tested GH5 endomannanases only reached 8-10% as their maximal degree of conversion. α-Galactosyl-mannose was identified as the dominant degradation product resulting from AnMan26A and PaMan26A action on guar gum, strongly indicating that these two enzymes can accommodate galactopyranosyl residues in the -1 and in the +1 subsite. The degradation of α-6(4)-6(3)-di-galactosyl-mannopentaose by AnMan26A revealed accommodation of galactopyranosyl residues in the -2, -1 and +1 subsite of the enzyme. Accommodation of galactopyranosyl residues in subsites -2 and +1 has not been observed for other characterized endomannanases to date. Docking analysis of galactomannooligosaccharides in available crystal structures and homology models supported the conclusions drawn from the experimental results. This newly discovered diversity of substrate degradation patterns demonstrates an expanded functionality of fungal endomannanases, than hitherto reported.

Biorefining of green biomasses to produce proteins for feed and food along with fibers/sugars for production of fuels and chemicals constitutes one viable pathway in the transition to a bio-economy. In this respect, maximizing annual biomass output is a key parameter and one potential new concept could be green harvest of cereal crops followed by early establishment of a cover crop. The objective of this study was to evaluate different scenarios for harvesting and biorefining triticale either at maturity or earlier in green growth stages. High-yielding triticale was grown under different regimes of N-application and was harvested at three stages of maturity from green to full maturity. Green biomass was fractionated into a fiber rich pulp and a protein rich juice, from which a protein concentrate was produced. The digestibility and sugar potential of the fiber pulp was evaluated against mature straw by enzymatic hydrolysis after hydrothermal pretreatment. Triticale biomass and protein yield did not increase during the last 5–6 weeks before harvest at maturity in August. On average 21 t ha−1 of dry weight biomass was harvested in early July. Fractionation of protein from the green triticale resulted in the production of 730 kg ha−1 of crude protein in a protein concentrate with an amino acid profile superior to grain protein. The structural carbohydrates in the green biomass and the fiber pulp were less recalcitrant compared to mature straw. The total sugar potential from biorefining of green triticale was under the given conditions up to 5.3 t ha−1.

Argumenter for fornuft og perspektiver i partnerskabelse mellem tre politiksystemer

Testing of cellulases on real biomass samples is required to do a true assessment of their efficacy for biomass degradation. Cellulase enzymes belong to a number of different glycosyl hydrolase families, all with different activity, specificity and modes of action. The concerted and synergistic action of these different cellulases determines the efficacy for plant cell wall deconstruction and cellulose hydrolysis. However, the plant cell wall of lignocellulosic materials is a very complex matrix and the efficacy of a cellulase preparation to degrade lignocellulosic materials is influenced by many factors. In this chapter, two protocols for testing efficacy of cellulases on pretreated biomass samples are described. The first protocol describes a small-scale setup employing low solids concentration that easily enables the testing of a larger number of samples. The second protocol describes a method for testing the efficacy of cellulases at conditions more closely resembling industrial conditions, i.e., high solids concentrations. Both protocols can be used to test the cellulases under a variety of substrate types, substrate concentrations, enzyme loadings and process conditions. The protocols can also be used to evaluate different feedstocks.

Bestemmelse af kollektivitet og solidaritet blandt danske lønmodtagere på baggrund af en større interviewundersøgelse

Since the beginning of the twenty-first century, Denmark’s flexicurity system has attracted attention from scholars and policy makers worldwide. By combining flexible labour markets, generous unemployment insurance and active labour market policies with strong emphasis on training, the Danish system produces institutional complementarities that underpin the competitiveness of firms without sacrificing goals of equality and social protection of workers. Moreover, the system facilitates ongoing coordination between employer and wage earner organisations. This means that challenges stemming from globalisation and technological changes can be tackled in an offensive manner and without the interference of the political system and yet still be legitimised on both sides of the class divide.

An integrated membrane system was investigated for the separation of mixtures of xylose and glucose. Separation of these sugars is extremely challenging due to their similar structure, size and charge. In order to enhance the xylose separation factor in nanofiltration (NF), we present an enzymatic process for converting glucose to gluconic acid followed by separation of xylose from gluconic acid by nanofiltration. Process conditions which favored the negative charge repulsions between gluconic acid and the NF270 membrane were examined. At the best conditions (9:1 feed molar ratio of xylose to gluconic acid, 0.15 M total feed concentration, pH 9.5, 25 °C and 4 bar), we achieved a xylose separation factor of 34 and a throughput of 18.7 L m −2 h −1 . In comparison, the separation factor was only 1.4 for solutions of xylose and glucose at the same process conditions, thus demonstrating the huge potential of the integrated system. Full conversion of glucose to gluconic acid assisted by glucose oxidase (GOD) could be achieved by coupling a parallel reaction catalyzed by catalase (CAT), where H 2 O 2 (GOD-inhibitor formed in the first reaction) was decomposed to water and oxygen. GOD has a high oxygen-demand and it was demonstrated that sufficient oxygen could be obtained by controlling the CAT-catalyzed reaction through initial H 2 O 2 addition. The new strategy suggested in this study, integrating reaction and nanofiltration to enhance separation while obtaining another value-added stream, presents new options for separating compounds with similar molecular weights by nanofiltration.

Background The amino acid profile of plants is an important parameter in assessments of their growth potential, resource-use efficiency and/or quality as food and feed. Screening studies may involve large number of samples but the classical amino acid analysis is limited by the fact that it is very time consuming with typical chromatographic run times of 70 min or more. Results We have here developed a high-throughput method for analysis of amino acid profiles in plant materials. The method combines classical protein hydrolysis and derivatization with fast separation by UHPLC and detection by a single quadrupole (QDa) mass spectrometer. The chromatographic run time is reduced to 10 min and the precision, accuracy and sensitivity of the method are in line with other recent methods utilizing advanced and more expensive mass spectrometers. The sensitivity of the method is at least a factor 10 better than that of methods relying on detection by fluorescence or UV. It is possible to downscale sample size to 20 mg without compromising reproducibility, which makes the method ideal for analysis of very small sample amounts. Conclusion The developed method allows high-throughput analysis of amino acid profiles in plant materials. The analysis is robust and accurate as well as compatible with both free amino acids and protein hydrolysates. The QDa detector offers high sensitivity and accuracy, while at the same time being relatively simple to operate and cheap to purchase, thus significantly reducing the overall analytical costs compared to methods based on more advanced mass spectrometers. Electronic supplementary material The online version of this article (10.1186/s13007-018-0277-8) contains supplementary material, which is available to authorized users.

Alkaline hydrothermal liquefaction (HTL) of lignin rich enzymatic hydrolysis residues (EnzHR) from wheat straw and Miscanthus x giganteus was performed at 255, 300, and 345 degrees C to investigate valorization of this side-stream from second-generation bioethanol production. The EnzHR were from biomass hydro thermally pretreated at two different levels of severity (190 degrees C, 10 min or 195 degrees C, 15 min), and HTL at 300 degrees C of these EnzHR showed the most effective lignin depolymerization of the low severity EnzHR for both wheat straw and Miscanthus. The degree of depolymerization during HTL was temperature dependent and was not complete after 20 min at 255 degrees C, most distinctly for the Miscanthus EnzHR. The yields of 128 monomeric products quantified by gas chromatography mass spectrometry were up to 15.4 wt % of dry matter. Principal component analysis of the quantified compounds showed that nonlignin HTL products are main contributors to the variance of the HTL products from the two biomasses. The chemically modified lignin polymer was found to have increased thermal stability after HTL. Analytical pyrolysis was applied to investigate the chemical composition of a larger fraction of the products. Analytical pyrolysis contributed with additional chemical information as well as confirming trends seen from quantified monomers. This work is relevant for future lignin valorization in biorefineries based on current second-generation bioethanol production.

Testing of cellulases on real biomass samples is required to do a true assessment of their efficacy for biomass degradation. Cellulase enzymes belong to a number of different glycosyl hydrolase families, all with different activity, specificity and modes of action. The concerted and synergistic action of these different cellulases determines the efficacy for plant cell wall deconstruction and cellulose hydrolysis. However, the plant cell wall of lignocellulosic materials is a very complex matrix and the efficacy of a cellulase preparation to degrade lignocellulosic materials is influenced by many factors. In this chapter, two protocols for testing efficacy of cellulases on pretreated biomass samples are described. The first protocol describes a small-scale setup employing low solids concentration that easily enables the testing of a larger number of samples. The second protocol describes a method for testing the efficacy of cellulases at conditions more closely resembling industrial conditions, i.e., high solids concentrations. Both protocols can be used to test the cellulases under a variety of substrate types, substrate concentrations, enzyme loadings and process conditions. The protocols can also be used to evaluate different feedstocks.

Analytisk indkredsning af social dumping og argumentation for kommuners opgaver i forbindelse med bekæmpelse heraf

Sugarcane bagasse, corn stover, and wheat straw are among the most available resources for the production of cellulosic ethanol. For these biomasses we study the influence of pre-treatment methods on the chemical composition, as well as on the subsequent reactions of enzymatic hydrolysis and oxidation of cellulose. The applied pre-treatment methods are organosolv, hydrothermal, and alkaline. Hydrothermally pretreated wheat straw gave the highest cellulose conversion with 80% glucose yield and 0.8% oxidized cellulose products. Recent studies have shown that lignin is able to boost the activity of the cellulose oxidizing enzyme lytic polysaccharide monooxygenase (LPMO). The highest activity of LPMO was observed for the hydrothermally pretreated biomasses, which also contained the highest level of lignin. All hydrolyses were done at high dry matter levels using a commercial enzyme preparation containing hydrolytic and oxidative enzymes.

We present a unique evaluation of three advanced high throughput pretreatment and enzymatic hydrolysis systems (HTPH-systems) for screening of lignocellulosic biomass for enzymatic saccharification. Straw from 20 cultivars of winter wheat from two sites in Denmark was hydrothermally pretreated and enzymatically processed in each of the

This article examines co-determination in the collective participation system in Denmark among semi-professional shop stewards and health and safety representatives in the private and the public sector. Union strategies for democratization of working life and economic democracy are no longer progressing as in the 1970s, and co-determination seems to be on the defensive, challenged by new forms of employer strategies, New Public Management principles and the neoliberal strategy for individualization. These developments call for a reappraisal of the capability of the system to secure influence and participation in decision-making. The main findings, based on comprehensive empirical investigations, are three-fold. First, a connection between new employer strategies and the level of influence and co-determination is recorded. Generally, the representatives find it harder to maintain co-determination in a change-intensive workplace context especially with regard to tactical and strategic decisions. Second, visions for democratization exist among the majority of the representatives, who wish to strengthen co-determination institutionally and at the strategic level. The obstacles call for new institutional rights of co-determination in the central cooperation agreement. Third, a broader upgrade of analytical skills is required to develop abilities to analyse and anticipate developments, to decode management strategies and see opportunities of change in the institution – a sort of politico-sociological imagination to make it possible to grasp the surrounding world’s signals as well as the opportunities to approach agenda-setting more proactively.

Biological degradation of biomass on an industrial scale culminates in high concentrations of end products. It is known that the accumulation of glucose and cellobiose, end products of hydrolysis, inhibit cellulases and decrease glucose yields. Aside from these end products, however, other monosaccharides such as mannose and galactose (stereoisomers of glucose) decrease glucose yields as well. NMR relaxometry measurements showed direct correlations between the initial T2 of the liquid phase in which hydrolysis takes place and the total glucose production during cellulose hydrolysis, indicating that low free water availability contributes to cellulase inhibition. Of the hydrolytic enzymes involved, those acting on the cellulose substrate, that is, exo- and endoglucanases, were the most inhibited. The β-glucosidases were shown to be less sensitive to high monosaccharide concentrations except glucose. Protein adsorption studies showed that this inhibition effect was most likely due to catalytic, and not binding, inhibition of the cellulases.

Fuels and chemicals derived through biochemical conversion of agricultural by-products such as wheat straw (Triticum aestivum L.) is an area currently under intense research. In this study, separate leaves and stems were hydrothermally pretreated and enzymatically hydrolysed and analysed chemically and by comprehensive microarray polymer profiling (CoMPP). This way, the effects of each degradation step to the intermolecular organisation of specific polysaccharides in the cell walls were elucidated. After pretreatment, the degree of polymerisation (DP) of released xylo-oligosaccharides in both samples was up to about 20, but mostly around 3–8, and notably more acetylated in stems. Arabinoxylan (AX) and mixed-linkage glucan (MLG) became water-extractable while xylan, xyloglucan (XG), mannan and glucan remained only alkali-extractable. All polysaccharides became partly digestible after pretreatment however, regardless their extractability in water or only alkali. Based on the results, AX and MLG appear to be loosely bound in the cell wall matrix while the other polysaccharides are bound more tightly and shielded from enzymatic attack by AX and MLG until pretreatment. The gradual solubilisation and digestion of the polysaccharides during pretreatment and hydrolysis correlate well with previous models of the polysaccharides’ structural organisation in the cell wall.

Biorefining of green biomass such as grass and grass-clover is an emerging field with extensive potentials for new value-chains and cascading use of biomasses. Grass biomass can be biorefined into a protein-rich juice fraction for monogastric feed purposes and a fibrous pulp fraction for ruminant feed or biogas production. This study investigated the ensiling of the pulp fraction after extraction of protein juice from two types of grass biomass, namely grass-clover and ryegrass with 28.8 and 33.6% total solid (TS) in the two pulp fractions, respectively. Lab-scale ensiling experiments for 3 to 5 months showed that increased time from processing of grass biomass to initiation of the ensiling of the pulp fraction resulted in higher final pH of the silage, particularly for the ryegrass pulp, and reduced concentration of lactic acid, total solid (TS), total amino acids (AAs) and free AAs. Addition of sucrose could, to some extent, counteract the effect of delayed ensiling on pH. Pilot-scale ensiling experiments for 7–11 months demonstrated limited effluent run-off with 1.8% for grass-clover pulp and 0.0% for ryegrass pulp. The ensiling of pulp had significant impact on a number of chemical constituents, e.g. with a reduction in neutral detergent fibre (NDF), in-vitro digestibility of organic matter (IVOS), total AAs and true protein but with an increase in free AAs. Also, ensiling significantly increased the biochemical methane potential (BMP) for anaerobic digestion durations up to 20 or 30 days but not at longer durations. Mass balances demonstrated a high recovery of volatile solids (97–98%), NDF (92–94%) and methane production potential (98–116%). However, recovery was lower for true protein (67–85%) and total AAs (84–90%), indicating some degradation of protein and AAs during ensiling. In conclusion, ensiling may offer an efficient way of conserving the quality of grass pulp, provided the process and logistics are optimized, particularly with a rapid sealing of the pulp after processing.

The role of endocellulases and endoxylanase during liquefaction and saccharification of hydrothermally pretreated wheat straw was studied. The use of a flow-loop setup with in-line magnetic resonance imaging enabled frequent measurements of viscosity at 55°C during saccharification at 6% total solids content. Viscosity data were complemented with off-line measurements of fiber lengths and release of soluble sugars. A clear correlation between fiber attrition and a decrease in viscosity was found. Fiber lengths and viscosity dropped quickly within the first hour and then stagnated, while sugar yields increased substantially thereafter, illustrating that liquefaction and saccharification are separate mechanisms. Both endoglucanase and endoxylanase were shown to have a significant effect on viscosity during liquefaction while the addition of endoxylanase also increased sugar yield.

Production of ethanol from lignocellulosic materials has a promising market potential, but the process is still only at pilot/demonstration scale due to the technical and economical difficulties of the process. Operating the process at very high solids concentrations (above 20% dry matter-DM) has proven essential for economic feasibility at industrial scale. Historically, simultaneous saccharification and fermentation (SSF) was found to give better ethanol yields compared to separate hydrolysis and fermentation (SHF), but data in literature are typically based on operating the process at low dry matter conditions. In this work the impact of selected enzyme preparation and processing strategy (SHF, presaccharification and simultaneous saccharification and fermentation-PSSF, and SSF) on final ethanol yield and overall performance was investigated with pretreated wheat straw up to 30% DM. The experiments revealed that an SSF strategy was indeed better than SHF when applying an older generation enzyme cocktail (Celluclast-Novozym 188). In case of the newer product Cellic CTec 2, SHF resulted in 20% higher final ethanol yield compared to SSF. It was possible to close the mass balance around cellulose to around 94%, revealing that the most relevant products could be accounted for. One observation was the presence of oxidized sugar (gluconic acid) upon enzymatic hydrolysis with the latest enzyme preparation. Experiments showed gluconic acid formation by recently discovered enzymatic class of lytic polysaccharides monoxygenases (LPMO's) to be depending on the processing strategy. The lowest concentration was achieved in SSF, which could be correlated with less available oxygen due to simultaneous oxygen consumption by the yeast. Quantity of glycerol and cell mass was also depending on the selected processing strategy.

Measurement of the protein content in samples from production of lignocellulosic bioethanol is an important tool when studying the adsorption of cellulases. Several methods have been used for this, and after reviewing the literature, we concluded that one of the most promising assays for simple and fast protein measurement on this type of samples was the ninhydrin assay. This method has also been used widely for this purpose, but with two different methods for protein hydrolysis prior to the assay-alkaline or acidic hydrolysis. In samples containing glucose or ethanol, there was significant interference from these compounds when using acid hydrolysis, which was not the case when using the alkaline hydrolysis. We evaluated the interference from glucose, cellulose, xylose, xylan, lignin and ethanol on protein determination of BSA, Accellerase(®) 1500 and Cellic(®) CTec2. The experiments demonstrated that the presence of cellulose, lignin and glucose (above 50 g/kg) could significantly affect the results of the assay. Comparison of analyses performed with the ninhydrin assay and with a CN analyser revealed that there was good agreement between these two analytical methods, but care has to be taken when applying the ninhydrin assay. If used correctly, the ninhydrin assay can be used as a fast method to evaluate the adsorption of cellulases to lignin.