Journal: Biotechnology for Biofuels

Article Title: The use of lytic polysaccharide monooxygenases in anaerobic digestion of lignocellulosic materials

doi: 10.1186/s13068-019-1611-8

Figure Lengend Snippet: Initial Glc4gemGlc concentrations in biogas reactions with Avicel supplemented with commercial cellulase cocktails (Cellic Ctec2 or Celluclast) or purified Nc LPMO9C as well as H 2 O 2

Article Snippet: Cellic ® CTec2 and Celluclast 1.5L were provided by Novozymes A/S (Bagsværd, Denmark).

Techniques: Purification, Control

Journal: Biotechnology for Biofuels

Article Title: The use of lytic polysaccharide monooxygenases in anaerobic digestion of lignocellulosic materials

doi: 10.1186/s13068-019-1611-8

Figure Lengend Snippet: Cumulative methane production during anaerobic digestion of Avicel with addition of Cellic Ctec2 ( a ), Celluclast ( b ), Nc LPMO9C ( c ) and an 85% Celluclast and 15% Nc LPMO9C (Cell + 9C) blend ( d ). The microbial inoculum was collected from a cow-manure-and-food-waste fed biogas reactor. Enzymes were supplied once at day 0 at 4 mg of protein per gram of substrate, with and without following addition of H 2 O 2 . Where indicated, hydrogen peroxide was supplied at 0 h, 24 h, 48 h and 72 h at 0.1 mM final concentration. Deionized water was added in all reactions without H 2 O 2 . Boiled enzyme control reactions with added H 2 O 2 are also shown. Background inoculum methane release was subtracted from all reactions in the calculation of methane production. The curves represent the average of two separate experiments. Methane production rates for these experiments are shown in Additional file : Figure S2

Article Snippet: Cellic ® CTec2 and Celluclast 1.5L were provided by Novozymes A/S (Bagsværd, Denmark).

Techniques: Concentration Assay, Control

Journal: Biotechnology for Biofuels

Article Title: The use of lytic polysaccharide monooxygenases in anaerobic digestion of lignocellulosic materials

doi: 10.1186/s13068-019-1611-8

Figure Lengend Snippet: Cumulative methane production during anaerobic digestion of birch with addition of Cellic Ctec2 ( a ), Celluclast ( b ), Nc LPMO9C ( c ) and an 85% Celluclast and 15% Nc LPMO9C (Cell + 9C) blend ( d ). The microbial inoculum was collected from a cow-manure-and-food-waste fed biogas reactor. Enzymes were supplied once at day 0 at 4 mg of protein per gram of substrate, with and without following addition of H 2 O 2 . Where indicated, hydrogen peroxide was supplied at 0 h, 24 h, 48 h and 72 h at 0.1 mM final concentration. Deionized water was added in all reactions without H 2 O 2 . Boiled enzyme control reactions with added H 2 O 2 are also shown. Inoculum methane release was subtracted from all samples in the calculation of methane production. The curves represent the average of two separate experiments. Methane production rates for these experiments are shown in Additional file : Figure S3

Article Snippet: Cellic ® CTec2 and Celluclast 1.5L were provided by Novozymes A/S (Bagsværd, Denmark).

Techniques: Concentration Assay, Control

Journal: Biotechnology for Biofuels

Article Title: The use of lytic polysaccharide monooxygenases in anaerobic digestion of lignocellulosic materials

doi: 10.1186/s13068-019-1611-8

Figure Lengend Snippet: Cumulative methane production during anaerobic digestion of spruce with addition of Cellic Ctec2 ( a ), Celluclast ( b ), Nc LPMO9C ( c ) and an 85% Celluclast and 15% Nc LPMO9C (Cell + 9C) blend ( d ). The microbial inoculum was collected from a cow-manure-and-food-waste fed biogas reactor. Enzymes were supplied once at day 0 at 4 mg of protein per gram of substrate, with and without following addition of H 2 O 2 . Where indicated, hydrogen peroxide was supplied at 0 h, 24 h, 48 h and 72 h at 0.1 mM final concentration. Deionized water was added in all reactions without H 2 O 2 . Boiled enzyme control reactions with added H 2 O 2 are also shown. Inoculum methane release was subtracted from all samples in the calculation of methane production. The curves represent the average of two separate experiments. Methane production rates for these experiments are shown in Additional file : Figure S4

Article Snippet: Cellic ® CTec2 and Celluclast 1.5L were provided by Novozymes A/S (Bagsværd, Denmark).

Techniques: Concentration Assay, Control

Journal: Biotechnology for Biofuels

Article Title: The use of lytic polysaccharide monooxygenases in anaerobic digestion of lignocellulosic materials

doi: 10.1186/s13068-019-1611-8

Figure Lengend Snippet: Cumulative methane production during anaerobic digestion of lignin-rich residue from birch (LRR) with addition of Cellic Ctec2 ( a ), Celluclast ( b ), Nc LPMO9C ( c ) and an 85% Celluclast and 15% Nc LPMO9C (Cell + 9C) blend ( d ). The microbial inoculum was collected from a cow-manure-and-food-waste fed reactor. Enzymes were supplied once at day 0 at 4 mg of protein per gram of substrate, with and without following addition of H 2 O 2 . Where indicated, hydrogen peroxide was supplied at 0 h, 24 h, 48 h and 72 h at 0.1 mM final concentration. Deionized water was added in all reactions without H 2 O 2 . Boiled enzyme control reactions with added H 2 O 2 are also shown. Inoculum methane release was subtracted from all samples in the calculation of methane production. The curves represent the average of two separate experiments. Methane production rates for these experiments are shown in Additional file : Figure S5

Article Snippet: Cellic ® CTec2 and Celluclast 1.5L were provided by Novozymes A/S (Bagsværd, Denmark).

Techniques: Residue, Concentration Assay, Control

Journal: Biotechnology for Biofuels

Article Title: The impact of hydrogen peroxide supply on LPMO activity and overall saccharification efficiency of a commercial cellulase cocktail

doi: 10.1186/s13068-018-1199-4

Figure Lengend Snippet: Effect of the concentrations of reductant and O 2 on saccharification of Avicel with Cellic ® CTec2. a , b Reactions under air saturated conditions (21% v/v O 2 ) using various concentrations of ascorbic acid (AscA). c , d Reactions with 5 mM AscA using various oxygen saturation levels in the headspace. The left panels ( a , c ) show production of Glc4gemGlc and the right panels ( b , d ) show glucan conversion. Reaction mixtures contained 10% (w/w) DM of Avicel and 4 mg/g DM of Cellic ® CTec2, in 50 mM sodium acetate buffer at pH 5.0 and were incubated at 50 °C. The error bars represent standard deviations for three independent experiments

Article Snippet: Controlled saccharification with continuous feeding of H 2 O 2 was conducted in 3 L glass bioreactors (Applikon, Schiedam, Netherlands) with 900 mL working volume, 10% (w/w DM) of cellulosic substrates and Cellic ® CTec2 (2 or 4 mg g −1 DM for Avicel, 4 mg g −1 DM for sulfite-pulped Norway spruce and 2 mg g −1 DM for less cellulose-rich SEB).

Techniques: Incubation

Journal: Biotechnology for Biofuels

Article Title: The impact of hydrogen peroxide supply on LPMO activity and overall saccharification efficiency of a commercial cellulase cocktail

doi: 10.1186/s13068-018-1199-4

Figure Lengend Snippet: Effect of various feeding regimes for H 2 O 2 and reductant (AscA) on Avicel oxidation by LPMOs present in Cellic ® CTec2. The various feeding scenarios are shown in the Table below the Figure. Note the positive control reaction (7, green: aerobic, 1 mM AscA, no H 2 O 2 added) and the two negative control reactions (6, gray: anaerobic, 1 mM AscA, no H 2 O 2 added; 5, purple: anaerobic, no AscA, with repetitive addition of 200 µM H 2 O 2 ). AscA and/or H 2 O 2 were added after 60, 120, 180 and/or 240 min as indicated in the Table. The blue dashed line represents the concentration of AscA (secondary y -axis) measured in reaction 1 (blue line; anaerobic, 1 mM AscA with repetitive addition of 200 µM H 2 O 2 ). The reactions mixtures contained 10% (w/w) DM of Avicel and 4 mg/g DM of Cellic ® CTec2, in 50 mM sodium acetate buffer at pH 5.0 and were incubated at 50 °C. The error bars represent standard deviations for three independent experiments

Article Snippet: Controlled saccharification with continuous feeding of H 2 O 2 was conducted in 3 L glass bioreactors (Applikon, Schiedam, Netherlands) with 900 mL working volume, 10% (w/w DM) of cellulosic substrates and Cellic ® CTec2 (2 or 4 mg g −1 DM for Avicel, 4 mg g −1 DM for sulfite-pulped Norway spruce and 2 mg g −1 DM for less cellulose-rich SEB).

Techniques: Positive Control, Negative Control, Concentration Assay, Incubation

Journal: Biotechnology for Biofuels

Article Title: The impact of hydrogen peroxide supply on LPMO activity and overall saccharification efficiency of a commercial cellulase cocktail

doi: 10.1186/s13068-018-1199-4

Figure Lengend Snippet: Effect of the H 2 O 2 feeding rate on the initial phase of saccharification of Avicel with Cellic ® CTec2. a , d The production of Glc4gemGlc, b , e The extent of glucan conversion and c , f the concentration of AscA. Unless otherwise stated, reactions were carried out with 10% (w/w) DM of Avicel, 4 mg/g DM of Cellic ® CTec2 and 1 mM of AscA, in 50 mM sodium acetate buffer at pH 5.0, at 50 °C. The reactions displayed in d – f were conducted with the following modifications (marked a , b or c ): a indicates the absence of AscA, b indicates the presence of 0.1 mM AscA, and c indicates the use of 2 mg/g DM of Cellic ® CTec2. H 2 O 2 was supplied at a constant flow rate of 600 µL h −1 using H 2 O 2 stock solutions with appropriate concentrations (see Table for details) to obtain the desired H 2 O 2 feeding rates (µM h −1 ). To ensure anaerobic conditions, all reactions were performed with constant sparging of nitrogen at a flow rate of 100 mL min −1 , with exception of the aerobic control reaction (labeled “O 2 ” ), which was sparged with air at the same flow rate. “N 2 ” stands for the anaerobic control reaction. Both control reactions were run without the addition of H 2 O 2 . Repeated addition of AscA to a final concentration of 1 mM, in the reaction fed with 3000 µM h −1 , is marked with a blue asterisk (*). Error bars for glucan conversion represent standard deviations of two technical replicates. The data depicted in a – c , except the data for the highest H 2 O 2 feeding rate, have been published previously

Article Snippet: Controlled saccharification with continuous feeding of H 2 O 2 was conducted in 3 L glass bioreactors (Applikon, Schiedam, Netherlands) with 900 mL working volume, 10% (w/w DM) of cellulosic substrates and Cellic ® CTec2 (2 or 4 mg g −1 DM for Avicel, 4 mg g −1 DM for sulfite-pulped Norway spruce and 2 mg g −1 DM for less cellulose-rich SEB).

Techniques: Concentration Assay, Sparging, Control, Labeling

Journal: Biotechnology for Biofuels

Article Title: The impact of hydrogen peroxide supply on LPMO activity and overall saccharification efficiency of a commercial cellulase cocktail

doi: 10.1186/s13068-018-1199-4

Figure Lengend Snippet: Effect of various constant and non-constant H 2 O 2 feeding rates on the saccharification of Avicel with Cellic ® CTec2. a , d The production of Glc4gemGlc, b , e the glucan conversion and c , f the concentration of AscA. Unless otherwise stated, reactions were carried out with 10% (w/w) DM of Avicel, 4 mg/g DM of Cellic ® CTec2 and 1 mM of AscA, in 50 mM sodium acetate buffer at pH 5.0 and 50 °C, with constant supply of H 2 O 2 (600 µL h −1 ). To ensure anaerobic conditions, all reactions were carried out with constant sparging of nitrogen at a flow rate of 100 mL min −1 , with the exception of the aerobic control reactions (O 2 and O 2 a ), which were sparged with air at the same flow rate. “N 2 ” stands for the anaerobic control reaction. All three control reactions were run without the addition of H 2 O 2 . “O 2 a ” was run in the absence of AscA. Repeated additions of AscA to a final concentration of 1 mM in the reactions fed at 300 and 600 µM h −1 are marked with an asterisk (*). For the experiment called “Decrease”, the H 2 O 2 feed rate was gradually lowered as follows: 300 µM h −1 from 0 to 6 h; 200 µM h −1 from 6 to 12 h; 100 µM h −1 from 12 to 24 h and 50 µM h −1 from 24 to 48 h. For the experiment called “Addition”, the H 2 O 2 feed rate was as follows: 0 µM h −1 from 0 to 24 h and 300 µM h −1 from 24 to 48 h. Error bars for glucan conversion represent standard deviations for two technical replicates. The decrease of Glc4gemGlc over time that is observed in some of the reactions ( a , d ) is due to a first order degradation process that is independent of the presence of AscA and H 2 O 2 ; see main text for details

Article Snippet: Controlled saccharification with continuous feeding of H 2 O 2 was conducted in 3 L glass bioreactors (Applikon, Schiedam, Netherlands) with 900 mL working volume, 10% (w/w DM) of cellulosic substrates and Cellic ® CTec2 (2 or 4 mg g −1 DM for Avicel, 4 mg g −1 DM for sulfite-pulped Norway spruce and 2 mg g −1 DM for less cellulose-rich SEB).

Techniques: Concentration Assay, Sparging, Control

Journal: Biotechnology for Biofuels

Article Title: The impact of hydrogen peroxide supply on LPMO activity and overall saccharification efficiency of a commercial cellulase cocktail

doi: 10.1186/s13068-018-1199-4

Figure Lengend Snippet: Effect of various constant H 2 O 2 feeding rates on the saccharification of sulfite-pulped Norway spruce with Cellic ® CTec2. a The production of Glc4gemGlc and b the glucan conversion. All reactions were carried out with 10% (w/w) DM of spruce, 4 mg/g DM of Cellic ® CTec2 and 1 mM of AscA, in 50 mM sodium acetate buffer at pH 5.0 and 50 °C with constant supply of H 2 O 2 (600 µL h −1 ) using H 2 O 2 stock solutions with appropriate concentrations (Table ). To ensure anaerobic conditions, all reactions were carried out with constant sparging of nitrogen at a flow rate of 100 mL min −1 , with the exception of the aerobic control reaction (O 2 ), which was sparged with air at the same flow rate. “N 2 ” stands for the anaerobic control reaction. Both control reactions were run without addition of H 2 O 2 . Error bars for glucan conversion represent standard deviations of two technical replicates

Article Snippet: Controlled saccharification with continuous feeding of H 2 O 2 was conducted in 3 L glass bioreactors (Applikon, Schiedam, Netherlands) with 900 mL working volume, 10% (w/w DM) of cellulosic substrates and Cellic ® CTec2 (2 or 4 mg g −1 DM for Avicel, 4 mg g −1 DM for sulfite-pulped Norway spruce and 2 mg g −1 DM for less cellulose-rich SEB).

Techniques: Sparging, Control

Journal: Biotechnology for Biofuels

Article Title: The impact of hydrogen peroxide supply on LPMO activity and overall saccharification efficiency of a commercial cellulase cocktail

doi: 10.1186/s13068-018-1199-4

Figure Lengend Snippet: Effect of various constant H 2 O 2 feeding rates on the saccharification of steam-exploded birch with Cellic ® CTec2. a The production of Glc4gemGlc and b the glucan conversion. All reactions were carried out with 10% (w/w) DM of birch, 2 mg/g DM of Cellic ® CTec2 in the absence of AscA, in 50 mM sodium acetate buffer at pH 5.0 and 50 °C with constant supply of H 2 O 2 (600 µL h −1 ) using H 2 O 2 stock solutions with appropriate concentrations (Table ). To ensure anaerobic conditions, all reactions were carried out with constant sparging of nitrogen at a flow rate of 100 mL min −1 , with the exception of the aerobic control reaction (O 2 ), which was sparged with air at the same flow rate. “N 2 ” and “N 2 Cys” stand for the anaerobic control reactions; the latter was carried out in the presence of 0.025% (w/v) l -cysteine hydrochloride monohydrate. All control reactions were run without addition of H 2 O 2 . Since the glucan fraction of this substrate was only 43.9%, here, in contrast to all other reactions reported in this study, Cellic ® CTec2 was dosed at 2 mg (rather than 4 mg) protein/g DM. Error bars for glucan conversion represent standard deviations of two technical replicates

Article Snippet: Controlled saccharification with continuous feeding of H 2 O 2 was conducted in 3 L glass bioreactors (Applikon, Schiedam, Netherlands) with 900 mL working volume, 10% (w/w DM) of cellulosic substrates and Cellic ® CTec2 (2 or 4 mg g −1 DM for Avicel, 4 mg g −1 DM for sulfite-pulped Norway spruce and 2 mg g −1 DM for less cellulose-rich SEB).

Techniques: Serial Time-encoded Amplified Microscopy, Sparging, Control