scholarly journals Biorefining: the role of endoglucanases in refining of cellulose fibers

Cellulose ◽  
2021 ◽  
Author(s):  
Martin Nagl ◽  
Oskar Haske-Cornelius ◽  
Lukas Skopek ◽  
Alessandro Pellis ◽  
Wolfgang Bauer ◽  
...  

AbstractWith an annual production of more than 400 million tons, paper is the main product of the largest biorefinery process industrially implemented. Enzymes have been used for pulp refining to dramatically reduce energy consumption. However, exact mechanisms related to the individual enzymes are hardly understood. Yet, this knowledge would be important to predict enzyme performance in industrial processes. Three commercial refining enzyme formulations showed different endoglucanase (1.25 nkat mg−1–13.7 nkat mg−1), β-glucosidase (0.57 nkat mg−1–1.34 nkat mg−1) and xylanase activities (1.78 nkat ml−1–62.1 nkat mg−1) on model substrates. Additionally, distinct amounts of reducing sugars from hardwood sulfate pulp were released. Endoglucases were purified from each formulation by using hydrophobic interaction and anion exchange chromatography and showed molecular weights from 20 to 55 kDa and specific activities ranging between 3.11 and 26.3 nkat mg−1 according to endoglucanase specific derivatized cellopentaose (CellG5). Refining trials of hardwood sulfate pulp were conducted using a PFI laboratory mill and fiber properties such as degree of refining or fiber length and properties of formed sheets like tensile index were monitored. Thereby, enzymes were dosed based on identical endoglucanase activity on CellG5. Enzyme formulations and purified endoglucanases led to an increase of the degree of refining of up to 47.9 [°SR] at 6000 PFI revolutions while the tensile index was improved by up to 76.0 Nm g−1. In summary, refining effects can be primarily attributed to endoglucanases indicating activity on CellG5 being a suitable parameter for enzyme dosing.

2020 ◽  
Author(s):  
Ian Sims ◽  
A Bacic

The soluble polymers secreted by cell-suspension cultures of Nicotiana plumbaginifolia contained 78% carbohydrate, 6% protein and 4% inorganic material. The extracellular polysaccharides were separated into three fractions by anion-exchange chromatography using a gradient of imidazole-HCl at pH 7 and the individual polysaccharides in each fraction were then isolated by selective precipitation and enzymic treatment. Monosaccharide and linkage compositions were determined for each polysaccharide after reduction of uronic acid residues and the degree of esterification of the various uronic acid residues in each polysaccharide was determined concurrently with the linkage types. Six components were identified: an arabinoxyloglucan (comprising 34% of the total polysaccharide) and a galactoglucomannan (15%) in the unbound neutral fraction, a type II arabinogalactan (an arabinogalactan-protein, 11%) and an acidic xylan (3%) in the first bound fraction, and an arabinoglucuronomannan (11%) and a galacturonan (26%) in the second bound fraction. © 1995.


Holzforschung ◽  
2017 ◽  
Vol 71 (10) ◽  
pp. 767-775 ◽  
Author(s):  
Dominic Lorenz ◽  
Ron Janzon ◽  
Bodo Saake

AbstractThe exact quantification of all carbohydrate constituents in wood and pulp is a challenge because of the various glycosidic linkages of the polysaccharides with different stabilities. The individual detector responses for the compounds in the hydrolysates additionally complicate the quantification as pure standards for 4-O-methyl-α-D-glucuronic acid (meGlcA) and related oligosaccharides are not commercially available for calibration. In the present paper, a new analytical procedure is presented, based on the reductive amination of the carbohydrates obtained via acidic and enzymatic hydrolysis of the polysaccharides before quantification by means of high performance anion exchange chromatography (HPAEC) and UV-detection. This approach was suitable for the analysis of neutral carbohydrates and uronic acids obtained via enzymatic hydrolysis from bleached pulps. In the case of unbleached pulps, the enzymatic hydrolysis was not complete and unhydrolyzed nano-scaled and micro-scaled particles remained in the hydrolysates as detected by dynamic light scattering (DLS) measurements. The new HPAEC-UV methodology was also applied to kraft pulps and a sulfite pulp; six different kinds of wood as well as wheat straw and bagasse. All relevant monosaccharides and the dimer of meGlcA and xylose could be detected in the hydrolysates. Accordingly, significantly higher yields of meGlcA were found compared to literature data.


1970 ◽  
Vol 26 ◽  
pp. 2-12
Author(s):  
Patricia Adamma Ekwumemgbo ◽  
James Adagadzu Kagbu ◽  
Andrew Jonathan Nok ◽  
Israel Kehinde Omoniyi ◽  
Paul Ocheme Ameh ◽  
...  

Plasma (100.00 ml) obtained from bovine blood by centrifugation was fractionatedinto seven precipitates by cold ethanol precipitation. The yield (amount) of proteins in theprecipitates calculated from the standard curve of BSA was 1160.83 mg, 806.57 mg,1149.94 mg, 8.79 mg, 19.88 mg, 21.98 mg, and 13.97 mg respectively, while the totalamount of protein obtained was 3180.00 mg. The precipitates were fractionated into 25fractions each by Anion Exchange Chromatography (AEC) and the amount of protein ineach fraction was obtained by Bradford protein assay. SDS-PAGE analysis was performedon the fractions with proteins and their estimated molecular weights were obtained with theaid of the molecular weight marker. The result indicated that cold ethanol precipitation andanion exchange chromatographic techniques are valuable tools for the purification ofbovine blood to obtain high grade α, β and γ-fibrinogen, IgM (μ-globulin), IgG (γ -globulin),alpha (α –globulin), β-globulin (E-globulin) and albumin.Keywords: Plasma; Chromatography; Electrophoresis; Albumin; Fibrinogen; GlobulinDOI: 10.3126/jncs.v26i0.3624Journal of Nepal Chemical SocietyVol. 26, 2010Page: 2-12


1978 ◽  
Vol 24 (8) ◽  
pp. 1305-1316 ◽  
Author(s):  
W D Bostick ◽  
S R Dinsmore ◽  
J E Mrochek ◽  
T P Waalkes

Abstract Soluble arylsulfatase (EC 3.1.6.1) is present in the body fluids of man in the form of two isoenzymes, arylsulfatase A and B, which reportedly are useful biochemical markers for certain types of malignancy. However, rapid assay of the individual isoenzymes is extremely difficult; procedures based on differential inhibition or activation of the isoenzymes in a mixture yield only semiquantitative results. A feature of these isoenzymes is their inhibition by some common anions (notably phosphate) at physiologic concentrations. The isoenzymes can be separated by anion-exchange chromatography, the B isoenzyme being eluted in the void volume and the A isoenzyme and the anionic inhibitors retarded. Lead is used to sequester phosphate, enabling measurement of A in the salt-eluted fraction. Using this technique, we have found significant elevations of B in the sera of patients with colorectal cancer. The potential of rapid, chromatographic separation coupled with continuous monitoring for arylsulfatase activity is discussed.


2006 ◽  
Vol 73 (1) ◽  
pp. 49-57 ◽  
Author(s):  
Ekaterini E Moschopoulou ◽  
Ioannis G Kandarakis ◽  
Efstathios Alichanidis ◽  
Emmanouil M Anifantakis

The objective of this work was to study the characteristics of the gastric aspartic proteinases chymosin and pepsin which are constituents of the kid rennet. The two enzymes were extracted from abomasal tissue of one kid from a local indigenous breed, separated from each other by DEAE-cellulose chromatography and then were purified by gel filtration and anion-exchange chromatography. The molecular weights of the purified kid chymosin and pepsin as determined by gel filtration were 36 kDa and 40 kDa respectively. The isoelectric point of kid chymosin was as multiple forms of 3–6 zones at pH 4·6–5·1, while that of kid pepsin was at pH [les ]3·0. Kid pepsin contained 0·37 molecules phosphorous per molecule and was totally inhibited by 5 μM pepstatin A, being more sensitive than kid chymosin. Both enzymes were almost equally as proteolytic as calf chymosin on total casein at pH 5·6. Kid pepsin activity was more pH and temperature dependent than kid chymosin activity. In comparison with the calf chymosin temperature sensitivity, the order of increased sensitivity was: calf chymosin <kid chymosin <kid pepsin.


2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 360-361 ◽  
Author(s):  
Tom N Dale ◽  
Tim Parr ◽  
Julie King ◽  
John Brameld

Abstract The study objective was to investigate the effects of in-vitro fibrolytic enzyme incubation on xylooligosaccharide (XOS) release from different wheat varieties. Six varieties of wheat were ground to a 300µm powder and subjected to total non-cellulosic hydrolysis with Trifluroacetic acid (TFA) and in-vitro incubations with one of three commercially available enzymes at their recommended doses: Econase XT (xylanase 160,000 BXU/g), Econase MP1000 (Mannase 100,000 MNU/g) or Econase BP700 (ß-glucanase 700,000 BU/g) or no added enzymes. Each wheat variety was suspended (at 5mg/ml) in 50mM sodium citrate buffer (pH 5.2) with or without the individual enzymes (n = 3), and incubated at 40.7°C in a shaking incubator. Samples were taken at 0 and 6hr. Concentrations of xylose, xylobiose and xylotriose were determined by High-Performance Anion-Exchange Chromatography coupled with Pulsed Electrochemical Detection fitted with a CarboPac PA20 (xylose) or CarboPac PA200 Column (Dionex, Thermo Scientific) and known xylose and XOS standards (Megazyme, Ltd). Data were analysed by one or two-way ANOVA (Genstat 19th Edition), with significance accepted at P &lt; 0.05. After TFA hydrolysis, there were significant differences between varieties (P &lt; .001, ANOVA) in total xylose contents (Huntsman &gt; Paragon &gt; Chinese spring &gt; Sinuelo &gt; Highbury = Pavon 74). In the in-vitro incubations, there was a significant effect of enzyme, but not wheat variety, on both xylose (P = 0.009) and xylotriose (P &lt; .001) release at 6 hours, with Econase XT releasing more than the other two enzymes. There was a significant enzyme x wheat variety interaction (P &lt; .001) for xylobiose release at 6 hours, with Econase MP1000 releasing the most from Huntsman. In conclusion, the 6 wheat varieties differed in their non-cellulosic xylose contents and there were clear differences in the amounts of XOS released by the 3 enzymes. Further trials are needed to investigate whether the in-vitro experiments are indicative of results observed in-vivo.


Parasitology ◽  
1990 ◽  
Vol 101 (3) ◽  
pp. 369-376 ◽  
Author(s):  
J. K. Stiles ◽  
G. A. Ingram ◽  
K. R. Wallbanks ◽  
D. H. Molyneux ◽  
I. Maudlin ◽  
...  

SUMMARYA midgut trypanolysin and an agglutinin from Glossina palpalis subspecies were isolated and partially characterized using anion-exchange chromatography and polyacrylamide gel electrophoresis. FPLC fractions of midgut extracts of Glossina palpalis palpalis caused agglutination and lysis of two trypanosome species (Trypanosoma congolense and Trypanosoma brucei brucei). although Glossina palpalis gambiensis caused only agglutination. The trypanolysin and agglutinin were active only in the posterior midguts, were heat labile above 50%C, had a periodic cycle of ‘activity’ in response to bloodmeal intake and were not affected by protease inhibitors or trypsin but were inactivated by pronase. The lytic substance contained two proteins with approximate molecular weights (Mr) of 12000 and 10000 Da respectively. The agglutinin had an approximate Mr of 67000 Da. Gamma-irradiation of the two subspecies caused a temporary inhibition of trypanolytic and agglutinin activities in midgut extracts.


Processes ◽  
2021 ◽  
Vol 9 (4) ◽  
pp. 709
Author(s):  
Shu-Hui Yeh ◽  
Wen-Kuang Hsu ◽  
Zi-Qing Chang ◽  
Sue-Hong Wang ◽  
Chang-Wei Hsieh ◽  
...  

Previous studies identified that extracts of Talinum triangulare rich in flavonoids and phenolic acids showed antioxidative and immunomodulatory activities. In this study, the L9 orthogonal array was used to determine the optimal extraction conditions for water-extracted polysaccharides of T. triangulare (TTP) by hot reflux extraction and ultrasonic assisted extraction (UAE) methods. Results showed that while both extraction methods obtained a maximum polysaccharide yield of 3.1%, the optimal conditions for obtaining TTP was by UAE method. TTP was separated into large (LTTP) and small (STTP) molecular weights by dialysis. Since LTTP showed better effects than STTP in inducing macrophages to produce nitric oxide (NO) and indirectly inhibiting human cervical cancer HeLa cells, six different LTTP fractions were separated using anion-exchange chromatography. Contents of polysaccharides, triterpenoids, polyphenols, and proteins and molecular weights of major polysaccharide in each fraction were analyzed. The F1 fraction of LTTP, which showed the highest inducing ability of mouse RAW264.7 macrophages to secrete NO and tumor necrosis factor-α, showed the most significant indirect inhibitory effect of human colon cancer SW620 cells. These results suggest that LTTP, especially the F1 fraction, of T. triangulare may be used in health foods or Chinese medicine for its immunomodulatory potential.


2020 ◽  
Author(s):  
Ian Sims ◽  
A Bacic

The soluble polymers secreted by cell-suspension cultures of Nicotiana plumbaginifolia contained 78% carbohydrate, 6% protein and 4% inorganic material. The extracellular polysaccharides were separated into three fractions by anion-exchange chromatography using a gradient of imidazole-HCl at pH 7 and the individual polysaccharides in each fraction were then isolated by selective precipitation and enzymic treatment. Monosaccharide and linkage compositions were determined for each polysaccharide after reduction of uronic acid residues and the degree of esterification of the various uronic acid residues in each polysaccharide was determined concurrently with the linkage types. Six components were identified: an arabinoxyloglucan (comprising 34% of the total polysaccharide) and a galactoglucomannan (15%) in the unbound neutral fraction, a type II arabinogalactan (an arabinogalactan-protein, 11%) and an acidic xylan (3%) in the first bound fraction, and an arabinoglucuronomannan (11%) and a galacturonan (26%) in the second bound fraction. © 1995.


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