Antonio L. Torres

TCF bleaching of wheat straw pulp using ozone and xylanase. Part A: paper quality assessment.

The XOAZRP TCF sequence was applied to bleach wheat straw pulp. Following each bleaching stage, the properties of the pulp (viz. kappa number, standard viscosity, borohydride viscosity and brightness) and of the resulting effluents were determined. The performance of the reagents was analyzed through the studies of xylanase treatment and crystallinity and scanning electron microscopy of the pulps. Finally, the pulp was refined at 1000 revolutions in a PFI mill and the mechanical properties of the resulting paper were determined and compared with those of paper from a eucalyptus pulp. Despite its shortcomings, wheat straw pulp can be effectively bleached with the proposed TCF sequence.

Molecular cloning and characterization of a multidomain endoglucanase from Paenibacillus sp BP-23: evaluation of its performance in pulp refining

Abstract The gene celB encoding an endoglucanase from Paenibacillus sp. BP-23 was cloned and expressed in Escherichia coli. The nucleotide sequence of a 4161 bp DNA fragment containing the celB gene was determined, revealing an open reading frame of 2991 nucleotides that encodes a protein of 106,927 Da. Comparison of the deduced amino acid sequence of endoglucanase B with known β-glycanase sequences showed that the encoded enzyme is a modular protein and exhibits high homology to enzymes belonging to family 9 cellulases. The celB gene product synthesized in E. coli showed high activity on carboxymethyl cellulose and lichenan while low activity was found on Avicel. Activity was enhanced in the presence of 10 mM Ca2+ and showed its maximum at 53 °C and pH 5.5. The effect of the cloned enzyme in modifying the physical properties of pulp and paper from Eucalyptus was tested (CelB treatment). An increase in mechanical strength of paper and a decrease in pulp dewatering properties were found, indicating that CelB treatment can be considered as a biorefining. Treatment with CelB gave rise to an improvement in paper strength similar to that obtained with 1,000 revolutions increase in mechanical refining. Comparison with the performances of recently developed endoglucanase A from the same strain and with a commercial cellulase showed that CelB produced the highest refining effect.

Effect of cellulase-assisted refining on the properties of dried and never-dried eucalyptus pulp

The effect of two different cellulases on the hornification phenomenon,in which drainability (Schopper–Riegler method) and mechanical propertiesdiminish when pulps are dried, was studied. The enzyme applications testedincluded a commercial enzyme named ComC (Pergalase A40 from CIBA) and alaboratory enzyme from Paenibacillus sp. strain BP-23namedCelB. Industrial never-dried Eucalyptus globulus bleachedkraft pulp was split in two halves and one of them was dried at ambientcontrolled conditions. We compared enzyme effects on both pulps (wet pulp anddried pulp) before and after PFI mill refining. Enzyme applications increaseddrainability (Schopper–Riegler method) and water retention value (WRV) ofnever-dried bleached pulp, although this did not imply an enhancement of themechanical properties of paper. Cellulase treatment of dried pulps, bycontrast,gave rise to increased drainability and WRV and also to improved mechanicalproperties. The changes caused by drying became less significant after enzymeapplication. Handsheets from CelB-treated dried pulps showed an improvement oftensile and burst indexes while tear decreased. The effect produced by CelB canbe considered a biorefining step. In fact, by means of enzyme treatment withCelB the properties of paper manufactured from dried pulp equalled theproperties attained from wet fibres, with the exception of tear index. Changeswere also found in surface fibre morphology, such as flakes and peeling due tocellulase treatment. The surface modification of fibres with cellulases givesrise to better bonding properties and a closer structure of paper. The finalconclusion is that treatment with cellulases could compensate the hornificationeffect and lead to an important saving of refining energy. The novel enzyme,CelB, was the most effective in improving paper properties and counterbalancingthe hornification effect caused by drying.

Effects of xylanase treatment on fibre morphology in totally chlorine free bleaching (TCF) of Eucalyptus pulp.

The effects of xylanase on pulp properties and fibre morphology in a TCF (totally chlorine free) bleaching sequence has been studied. Enzymatic treatment is usually applied after oxygen delignification, but in this paper the effect of xylanase treatment before the oxygen delignification bleaching stage was studied. The application of xylanases in TCF bleaching of Eucalyptus kraft pulps enables the production of fully bleached pulps with high brightness (90% ISO) and viscosity (above 800 mL:g). The optimal NaOH charge in the oxygen delignification stage was studied. Xylanase treatment improves the accessibility of the bleaching chemicals to the fibres. The improvement of ‘bleachability’ produced by the enzyme reduces the quantity of ozone needed in subsequent bleaching stages. SEM (scanning electron microscopy) performed after oxygen delignification showed notable changes on fibre surfaces.

Engineering a family 9 processive endoglucanase from Paenibacillus barcinonensis displaying a novel architecture

Cel9B from Paenibacillus barcinonensis is a modular endoglucanase with a novel molecular architecture among family 9 enzymes that comprises a catalytic domain (GH9), a family 3c cellulose-binding domain (CBM3c), a fibronectin III-like domain repeat (Fn31,2), and a C-terminal family 3b cellulose-binding domain (CBM3b). A series of truncated derivatives of endoglucanase Cel9B have been constructed and characterized. Deletion of CBM3c produced a notable reduction in hydrolytic activity, while it did not affect the cellulose-binding properties as CBM3c did not show the ability to bind to cellulose. On the contrary, CBM3b exhibited binding to cellulose. The truncated forms devoid of CBM3b lost cellulose-binding ability and showed a reduced activity on crystalline cellulose, although activity on amorphous celluloses was not affected. Endoglucanase Cel9B produced only a small ratio of insoluble products from filter paper, while most of the reducing ends produced by the enzyme were released as soluble sugars (91%), indicating that it is a processive enzyme. Processivity of Cel9B resides in traits contained in the tandem of domains GH9–CBM3c, although the slightly reduced processivity of truncated form GH9–CBM3c suggests a minor contribution of domains Fn31,2 or CBM3b, not contained in it, on processivity of endoglucanase Cel9B.

Effect of a novel enzyme on fibre morphology during ECF bleaching of oxygen delignified Eucalyptus kraft pulps.

The use of a novel ‘‘xylanase A’’ enzyme was found beneficial in the elemental chlorine free (ECF) bleaching of oxygen delignified Eucalyptus kraft pulps. The application of the enzyme made possible production of fully bleached pulps with high brightness (89% ISO) and viscosities (above 800 cm 3 /g), without elemental chlorine (XDnEpD and DnEpD sequences) at low chlorine dioxide consumption. As a result of the enzyme treatment, morphological changes such as cracks, flakes, filaments and peeling of the fibre cell wall were observed by SEM (scanning electron microscopy). These modifications of the fibre surface facilitated the access to the fibre cell wall for the successive bleaching with less chemicals. ” 2000 Elsevier Science Ltd. All rights reserved.

Cloning of a new endoglucanase gene from Bacillus sp. BP-23 and characterisation of the enzyme. Performance in paper manufacture from cereal straw

Abstract The gene celA, encoding an endoglucanase from the strain Bacillus sp. BP-23, was cloned and expressed in Escherichia coli. The nucleotide sequence of a 1867-bp DNA fragment containing the celA gene was determined, revealing an open reading frame of 1200 nucleotides that encodes a protein of 44 803 Da. The deduced amino acid sequence of the encoded enzyme shows high homology to those of enzymes belonging to subtype 4 of the family-A cellulases. The celA gene product synthesized in E. coli showed activity on carboxymethylcellulose and lichenan but no activity was found on Avicel. Activity was enhanced in the presence of 10 mM Mg2+ and Ca2+ and showed its maximum at 40 °C and pH 4.0. Study of the performance of CelA on paper manufacture from agricultural fibres showed that treatment with the enzyme improved the properties of the pulp and the quality of paper. CelA treatment enhanced the physical properties (stretch and tensile index) of paper from wheat straw, while dewatering properties were slightly diminished. Electron-microscope analysis showed that the surface of straw fibres was modified by CelA.

Influence of the hexenuronic acid content on refining and ageing in eucalyptus TCF pulp

Brightness in totally chlorine-free (TCF) pulp is more unstable than it is in elemental chlorine-free (ECF) pulp, seemingly by effect of the former containing greater amounts of oxidizable structures such as hexenuronic acids (HexA). Accelerated ageing tests involving the application of moist heat to pulp revealed that brightness reversion can be alleviated by using an effective biotechnological method involving an enzyme-mediator treatment. Thus, the joint use of laccase from Trametes villosa and the mediator hydroxybenzotrialoze (HBT) in TCF pulp removes hexenuronic acids by 23% and reduces brightness reversion by 8.4%. Additional tests conducted to assess the effect of HexA on pulp refining revealed that these oxidizable structures introduce hydrophilicity in the pulp. Removing hexenuronic acids from TCF pulp alters its refining outcome as regards drainability ( degrees SR) and water retention capacity (%WRV); also, it leads to paper with comparable strength-related properties which requires no additional refining energy.

Tcf bleaching of wheat straw pulp using ozone and xylanase. Part B: kinetic studies

The reaction kinetics of ozone bleaching of wheat straw pulp has been studied for the first time. The results were compared with eucalyptus pulp in order to know that both raw materials have a similar behaviour. Ozone treatments were carried out in a special reactor at low consistency (0.5% o.d.p.). The main variables were consumption of ozone by the pulp and application of a xylanase treatment (X) prior to the oxygen stage (O). The responses measured were kappa number, viscosity and brightness, to give the kinetic expressions for delignification, cellulose degradation and elimination of chromophore groups, along with calculation of selectivity. Cellulose degradation and elimination of lignin and chromophore groups show first-order kinetics in all cases. The kinetics of the enzyme pre-treatment effect shows similar behaviour in both raw materials, although the constants of delignification and elimination of chromophore are higher in straw pulp.

Increasing yield of nanocrystalline cellulose preparation process by a cellulase pretreatment

In this work the introduction of a cellulase treatment prior to NCC isolation was assessed. NCC was produced using sulfuric acid at two different concentrations (62 and 64% wt.). The effect of pore size for filtration step was also assessed. The smaller acid dose leaded to yields up to 65-70% and average size up to 160 nm. It also produced crystals with reduced sulfur content (0.6-1%). Cellulase pretreatment influenced NCC characteristics, as it increased overall yield a 12%, increased average particle size around 35 nm and reduced NCC sulfur content up to a 0.8%. We found that different conditions of enzymatic treatments led to quantitative differences on their effects on NCC. Acetate buffer used for enzymatic treatments was found to counteract effects of acid. The evidence presented in this work suggested that pretreating fibers with this cellulase represents a very interesting option to partially replace chemicals on NCC isolation.