Using in situ nanocellulose‐coating technology based on dynamic bacterial cultures for upgrading conventional biomedical materials and reinforcing nanocellulose hydrogels

Bacterial nanocellulose (BNC) is a microbial nanofibrillar hydrogel with many potential applications. Its use is largely restricted by insufficient strength when in a highly swollen state and by inefficient production using static cultivation. In this study, an in situ nanocellulose‐coating technology created a fabric‐frame reinforced nanocomposite of BNC hydrogel with superior strength but retained BNC native attributes. By using the proposed technology, production time could be reduced from 10 to 3 days to obtain a desirable hydrogel sheet with approximately the same thickness. This novel technology is easier to scale up and is more suitable for industrial‐scale manufacture. The mechanical properties (tensile strength, suture retention strength) and gel characteristics (water holding, absorption and wicking ability) of the fabric‐reinforced BNC hydrogel were investigated and compared with those of ordinary BNC hydrogel sheets. The results reveal that the fabric‐reinforced BNC hydrogel was equivalent with regard to gel characteristics, and exhibited a qualitative improvement with regard to its mechanical properties. For more advanced applications, coating technology via dynamic bacterial cultures could be used to upgrade conventional biomedical fabrics, i.e. medical cotton gauze or other mesh materials, with nanocellulose. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1077–1084, 2016     

Biocompatibility evaluation of densified bacterial nanocellulose hydrogel as an implant material for auricular cartilage regeneration

Bacterial nanocellulose (BNC), synthesized by the bacterium Gluconacetobacter xylinus, is composed of highly hydrated fibrils (99 % water) with high mechanical strength. These exceptional material properties make BNC a novel biomaterial for many potential medical and tissue engineering applications. Recently, BNC with cellulose content of 15 % has been proposed as an implant material for auricular cartilage replacement, since it matches the mechanical requirements of human auricular cartilage. This study investigates the biocompatibility of BNC with increased cellulose content (17 %) to evaluate its response in vitro and in vivo. Cylindrical BNC structures (Ø48?×?20 mm) were produced, purified in a built-in house perfusion system, and compressed to increase the cellulose content in BNC hydrogels. The reduction of endotoxicity of the material was quantified by bacterial endotoxin analysis throughout the purification process. Afterward, the biocompatibility of the purified BNC hydrogels with cellulose content of 17 % was assessed in vitro and in vivo, according to standards set forth in ISO 10993. The endotoxin content in non-purified BNC (2,390 endotoxin units (EU)/ml) was reduced to 0.10 EU/ml after the purification process, level well below the endotoxin threshold set for medical devices. Furthermore, the biocompatibility tests demonstrated that densified BNC hydrogels are non-cytotoxic and cause a minimal foreign body response. In support with our previous findings, this study concludes that BNC with increased cellulose content of 17 % is a promising non-resorbable biomaterial for auricular cartilage tissue engineering, due to its similarity with auricular cartilage in terms of mechanical strength and host tissue response.     

Melanoma growth in non-chemically modified translucid bacterial nanocellulose hollow and compartimentalized spheres

BackgroundBacterial nanocellulose (BNC) has been used as cell support in numerous tissue engineering studies. Its use can be explained based on the fact its structure allows the creation of a required microenvironment for an ideal material, which supports 3D cell culture. Its structure and interconnected pores lead to animal cells adhesion and proliferation, also allowing oxygen and nutrients transportation.MethodsWe developed a new methodology to produce spherical platforms synthesized by Komagataebacter hansenii (ATCC 23769) under dynamic culture conditions in minimal medium. The chemical composition and physical properties of the platforms were evaluated. Then, human melanoma cells (SK-MEL-28) were encapsulated into the platforms and evaluated by metabolic activity, morphology and their ability on adhering to the Hollow Translucid BNC Spheres (BNC-TS-H) and Compartmentalized Translucid BNC Spheres (BNC-TS-C) up to 3 days.ResultsBNC-TS-H and BNC-TS-C platforms were produced as translucid spheroid platforms with distinct microenvironment under dynamic fermentation. The chemical and physical characterizations confirmed the platforms composition as BNC. The produced internal microenvironments in spherical platforms are relevant to determine tumor cell fate. In the first 12 h of culture, cells could adhere to nanocellulose microfibers assuming their typical tumorous phenotype in 72 h of culture.ConclusionThe dynamic fermentation in minimal medium produced distinct microstructured platforms of BNC-TS-H and BNC-TS-C. The platforms microstructure resulted in microenvironments that enabled distinct cell-cell and cell-matrix interactions. This behavior suggests several applications in tissue engineering.General significanceThe method produced translucid BNC sphere platforms with distinct microenvironments for 3D cell culture.     

White biotechnology for cellulose manufacturing—The HoLiR concept

A variety of approaches are available for generation of bacteria‐produced nanocellulose (BNC) in different forms. BNC production under static cultivation conditions usually results in fleeces or foils, characterized by a homogeneous, three‐dimensional network of nanofibers and a uniform surface. However, under static cultivation conditions in batch vessels, the widths and the lengths of the BNC sheets cultured are determined by the dimensions of the culture vessel. In this contribution, a novel, efficient process for a (semi‐)continuous cultivation of planar BNC fleeces and foils with a freely selectable length and an adjustable height is presented. By means of comprehensive investigations, the comparability of the BNC harvested to that gained from static cultivation under batch conditions is demonstrated. A first estimation of the production costs further shows that this type of processing allows for significant cost reductions compared to static cultivation of BNC in Erlenmeyer flasks. Biotechnol. Bioeng. 2010. 105: 740–747. © 2009 Wiley Periodicals, Inc.     

Molecular aspects of bacterial nanocellulose biosynthesis

Bacterial nanocellulose (BNC) produced by aerobic bacteria is a biopolymer with sophisticated technical properties. Although the potential for economically relevant applications is huge, the cost of BNC still limits its application to a few biomedical devices and the edible product Nata de Coco, made available by traditional fermentation methods in Asian countries. Thus, a wider economic relevance of BNC is still dependent on breakthrough developments on the production technology. On the other hand, the development of modified strains able to overproduce BNC with new properties – e.g. porosity, density of fibres crosslinking, mechanical properties, etc. – will certainly allow to overcome investment and cost production issues and enlarge the scope of BNC applications. This review discusses current knowledge about the molecular basis of BNC biosynthesis, its regulations and, finally, presents a perspective on the genetic modification of BNC producers made possible by the new tools available for genetic engineering.     

Improved bacterial nanocellulose production from glucose without the loss of quality by evaluating thirteen agitator configurations at low speed

Thirteen agitator configurations were investigated at low speed in stirred-tank reactors (STRs) to determine if improved crude bacterial nanocellulose (BNC) productivity can be achieved from glucose-based media while maintaining high BNC quality using Komagataeibacter xylinus ATCC 23770 as a model organism. A comparison of five single impellers showed the pitched blade (large) was the optimal impeller at 300 rpm. The BNC production was further increased by maintaining the pH at 5.0. Among the single helical ribbon and frame impellers and the combined impellers, the twin pitched blade provided the best results. The combined impellers at 150 rpm performed better than the single impellers, and after optimizing the agitation conditions, the twin pitched blade (large) and helical ribbon impellers performed the best at 100 rpm. The performances of different agitators at low speed during BNC production were related to how efficiently the agitators improved the oxygen mass transfer coefficient. The twin pitched blade (large) was verified as providing the optimum performance by an observed crude BNC production of 1.97 g (L×d)⁻¹ and a BNC crude yield of consumed glucose of 0.41 g g⁻¹, which were 2.25 and 2.37 times higher than the initial values observed using the single impeller respectively. Further characterization indicated that the BNC obtained at 100 rpm from the STR equipped with the optimal agitator maintained high degree of polymerization and crystallinity.     

Comparison of productivity and quality of bacterial nanocellulose synthesized using culture media based on seven sugars from biomass

Komagataeibacter xylinus ATCC 23770 was statically cultivated in eight culture media based on different carbon sources, viz. seven biomass-derived sugars and one sugar mixture. The productivity and quality of the bacterial nanocellulose (BNC) produced in the different media were compared. Highest volumetric productivity, yield on consumed sugar, viscometric degree of polymerization (DP_v, 4350–4400) and thermal stability were achieved using media based on glucose or maltose. Growth in media based on xylose, mannose or galactose resulted in lower volumetric productivity and DP_v, but in larger fibril diameter and higher crystallinity (76–78%). Growth in medium based on a synthetic sugar mixture resembling the composition of a lignocellulosic hydrolysate promoted BNC productivity and yield, but decreased fibril diameter, DP_v, crystallinity and thermal stability. This work shows that volumetric productivity, yield and properties of BNC are highly affected by the carbon source, and indicates how industrially relevant sugar mixtures would affect these characteristics.     

Comparison of tolerance of four bacterial nanocellulose-producing strains to lignocellulose-derived inhibitors

Background

Through pretreatment and enzymatic saccharification lignocellulosic biomass has great potential as a low-cost feedstock for production of bacterial nanocellulose (BNC), a high value-added microbial product, but inhibitors formed during pretreatment remain challenging. In this study, the tolerance to lignocellulose-derived inhibitors of three new BNC-producing strains were compared to that of Komagataeibacter xylinus ATCC 23770. Inhibitors studied included furan aldehydes (furfural and 5-hydroxymethylfurfural) and phenolic compounds (coniferyl aldehyde and vanillin). The performance of the four strains in the presence and absence of the inhibitors was assessed using static cultures, and their capability to convert inhibitors by oxidation and reduction was analyzed.

Results

Although two of the new strains were more sensitive than ATCC 23770 to furan aldehydes, one of the new strains showed superior resistance to both furan aldehydes and phenols, and also displayed high volumetric BNC yield (up to 14.78 ± 0.43 g/L) and high BNC yield on consumed sugar (0.59 ± 0.02 g/g). The inhibitors were oxidized and/or reduced by the strains to be less toxic. The four strains exhibited strong similarities with regard to predominant bioconversion products from the inhibitors, but displayed different capacity to convert the inhibitors, which may be related to the differences in inhibitor tolerance.

Conclusions

This investigation provides information on different performance of four BNC-producing strains in the presence of lignocellulose-derived inhibitors. The results will be of benefit to the selection of more suitable strains for utilization of lignocellulosics in the process of BNC-production.

     

Purification and characterization of a psychrophilic catalase from Antarctic Bacillus

Catalase from Bacillus sp. N2a (BNC) isolated from Antarctic seawater was purified to homogeneity. BNC has a molecular mass of about 230 kDa and is composed of four identical subunits of 56 kDa. The catalase showed optimal activity at 25 degrees C and at a pH range of 6-11. The enzyme could be inhibited by azide, hydroxylamine, and mercaptoethanol. These characteristics suggested that BNC is a small-subunit monofunctional catalase. The activation energy of BNC was 13 kJ/mol and the apparent kcat/Km values were 3.6 x 10(6) and 4 x 10(6) L.mol(-1).s(-1) at 4 and 25 degrees C, respectively. High catalytic efficiency of BNC at low temperatures enables this bacterium to scavenge H2O2 efficiently. BNC exhibited activation energy, catalytic efficiency, and thermostability comparable with some mesophilic homologues. Such similarity of enzymatic characteristics to mesophilic homologues, although uncommon among the cold-adapted enzymes in general, has also been observed in other psychrophilic small-subunit monofunctional catalases.     

CALU-3 lung cells three-dimensionally assembled onto CellFate® matrix present angiotensin-converting enzyme-2 activity

Currently, there is a great need for the development of three-dimensional (3D) in vitro lung models. Particularly, the production of a suitable 3D model of pulmonary epithelium for understanding the pathophysiology of diseases such as the COVID-19 must consider the tissue architecture and presence, for example, of the angiotensin-converting enzyme-2 (ACE-2) in the cells. Different polymeric membranes are being used to support cell culturing, especially of lung cells, however, there is still no information about the culture of these cells onto bacterial nanocellulose (BNC) matrices. We have used the BNC matrix CellFate® as a support for the assembly of a 3D in vitro model of lung epithelium, composed of human lung fibroblasts (HLF) and lung adenocarcinoma cells (CALU-3). CellFate® matrices were made from bacterial fermentation resulting in a natural and biocompatible biopolymer. Cells were cultured onto CellFate® and maintained in a 5% CO₂ humidified atmosphere at 37°C. Cell viability was assessed by the resazurin method The samples were, then, exposed to the air–liquid interface (ALI), and histologically analyzed. ACE-2 activity was verified on the hydrolyze of the fluorogenic substrate Mca-APK(Dnp)-OH, and its presence was evaluated by flow cytometry. The expression of the anionic transporter SLCO3A1 was evaluated by qPCR. Cell viability analysis indicates that CellFate® was not toxic to these cells. By flow cytometry, the presence of the ACE-2 was identified in the CALU-3 cells surface corroborating the results obtained from enzymatic activity analysis. The SLCO3A1 transporter expression was identified in cells cultured onto CellFate®, but not in cells cultured onto the transwell (control). CALU-3 cells cultivated onto CellFate® resulted in a pseudostratified organization, a typical morphology of the human respiratory tract epithelium. The current model opens perspectives for studies involving physiological characterization, improving its relevance for the understanding of the pathophysiology of diseases as well as the response to drugs.     

Production of nanocellulose by enzymatic hydrolysis: Trends and challenges

There is a great interest in increasing the levels of production of nanocellulose, either by adjusting production systems or by improving the raw material. Despite all the advantages and applications, nanocellulose still has a high cost compared to common fibers and to reverse this scenario the development of new, cheaper, and more efficient means of production is required. The market trend is to have an increase in the mass production of nanocellulose; there is a great expectation of world trade. In this sense, research in this sector is on the rise, because once the cost is not an obstacle to production, this material will have more and more market. Production of the cellulose fibers is determinant for the production of nanocellulose by a hydrolyzing agent with a reasonable yield. This work presents several aspects of this new material, mainly addressing the enzymatic pathway, presenting the hydrolysis conditions such as pH, biomass concentration, enzymatic loading, temperature, and time. Also, the commonly used characterization methods are presented, as well as aspects of the nanocellulose production market.     

Chelating agents improve enzymatic solubilization of pectinaceous co-processing streams

This study investigates the hypothesis that loosening of the egg-box structure by presence of divalent ion chelating agents during enzymatic degradation of homogalacturonan (HG) can improve enzymatic polysaccharide solubilization on pectinaceous, agro-industrial co-processing streams. The influence of different levels of ethylene-diaminetetraacetic acid (EDTA), citric acid, oxalic acid, and phosphate was assessed in relation to enzymatic solubilization of isopropanol precipitatable oligo- and polysaccharides from sugar beet pulp, citrus peel, and two types of potato pulp. The two types of potato pulp were FiberBind 400, a dried commercial potato pulp product, and PUF, a dried calcium reduced product, respectively. The enzymatic treatment consisted of 1% (w/w) of substrate treated with pectin lyase from Aspergillus nidulans and polygalacturonase from A. aculeatus [each dosed at 1.0% (w/w) enzyme/substrate] at 60 °C, pH 6.0 for 1 min. Characterization of the released fractions demonstrated a significantly improved effect of chelating agents for polysaccharide solubilization from FiberBind 400, PUF, and citrus peel, whereas only low amounts of polysaccharides were solubilized from the sugar beet pulp. The results substantiated the importance of chelating agents during enzymatic extraction of pectinaceous polysaccharides. Lower levels of chelating agents were required for the calcium-reduced potato pulp substrate (PUF) indicating the significance of calcium cross-linking in HG in relation to the enzymatic solubilization yields. The effect of the chelating agents correlated to their dissociation constants (pK_a values) and calcium binding constants and citric acid and EDTA exerted highest effects. Maximum polysaccharide yield was obtained for FiberBind 400 where the enzymatic treatment in presence of citric acid yielded 22.5% (w/w) polysaccharides of the initial substrate dry matter.     

Laccase for biobleaching of eucalypt kraft pulp by means of a modified industrial bleaching sequence

Biobleaching of kraft pulp is a possible application of laccase, but it has not been described in detail for complete industrial bleaching sequences yet. Therefore, in this work, the biobleaching of Eucalyptus globulus kraft pulp was performed using a modified industrial totally chlorine‐free sequence. The modification consisted in the substitution of an enzymatic delignification stage, based on the application of laccase from Trametes villosa, for the first alkaline extraction one. The enzymatic stage was performed with several synthetic and natural mediators, namely 1‐hydroxybenzotriazole (HBT), violuric acid (VA), methyl syringate, and syringaldehyde. Several pulp properties were analyzed after each stage of the bleaching process—kappa number, ISO brightness, viscosity, and optical properties of CIEL*a*b* system. The new biobleaching sequence improved the pulp properties, in comparison to the conventional bleaching sequence, if HBT or VA was used as mediators. VA was selected as the best mediator of those tested and the effect of its concentration in the enzymatic stage was subsequently studied. Reducing the initial concentration by 30%, the same pulp quality was obtained, but if the reduction attained 60%, an important decrease in pulp integrity was detected. The modified bleaching sequence could improve the bleached pulp properties (kappa number 10%, ISO brightness 1%, and viscosity 5%) in comparison to the mill sequence. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012     

Statistically designed optimisation of enzyme catalysed starch removal from potato pulp

Potato pulp is a high-volume, low-value byproduct stream resulting from the industrial manufacture of potato starch. The pulp is a rich source of biologically functional dietary fibers, but the targeted valorisation of the fibers requires removal of the residual starch from the pulp. The objective of this study was to release the residual starch, making up 21–22% by weight of the dry matter, from the potato pulp in a rational way employing as few steps, as few enzyme activities, as low enzyme dosages, as low energy input (temperature and time), and as high pulp dry matter as possible. Starch removal to obtain dietary fibers is usually accomplished via a three step, sequential enzymatic treatment procedure using a heat stable α-amylase, protease, and amyloglucosidase. Statistically designed experiments were performed to investigate the influence of enzyme dose, amount of dry matter, incubation time and temperature on the amount of starch released from the potato pulp. The data demonstrated that all the starch could be released from potato pulp in one step when 8% (w/w) dry potato pulp was treated with 0.2% (v/w) (enzyme/substrate (E/S)) of a thermostable Bacillus licheniformis α-amylase (Termamyl^® SC) at 70 °C for at least 65 min. The study also indicated that the amount of other carbohydrates released from the pulp during the release of starch was less than using the AOAC Official Method 985.29 and another recently published starch release method employed as a pretreatment for enzymatic upgrading of a pectinaceous potato pulp fiber.     

菊芋粕菊粉的酶法提取、组成及抗氧化活性研究

本研究拟应用酶法提取技术解决菊芋菊粉工业化生产中的废弃物——菊芋粕再利用程度低的问题,并评价菊芋粕菊粉的抗氧化活性功效。对菊芋粕菊粉的果胶酶酶法提取的最佳条件通过响应面法进行了优化,并对菊芋初次水提菊粉(primary water-extracted inulin,PWI)和二次酶提菊粉(secondary enzymatic-extracted inulin,SEI)的组成成分和抗氧化活性进行了比较分析。响应面法优化确定的菊芋粕菊粉最佳酶法提取条件为:pH4.5、提取温度50 ℃、酶底比7.5 U/g、提取时间2 h,该提取方法所得菊芋粕菊粉的得率为35.30%±0.85%,与传统热水浸提法相比菊粉得率提高38.16%。组成分析结果显示,SEI的总糖和菊粉含量均显著高于PWI( P <0.05)。在菊粉聚合度方面,PWI中蔗果三糖和蔗果四糖含量较高,而SEI中蔗果五糖、蔗果六糖及其以上聚合度菊粉的含量较高。此外,SEI的抗氧化活性优于PWI。因此,果胶酶辅助提取方法有望为菊芋粕菊粉的再利用难题提供新的解决思路。     

Enzymatic hydrolysis and simultaneous saccharification and fermentation of alkali/peracetic acid-pretreated sugarcane bagasse for ethanol and 2,3-butanediol production

The enzymatic digestibility of alkali/peracetic acid (PAA)-pretreated bagasse was systematically investigated. The effects of initial solid consistency, cellulase loading and addition of supplemental β-glucosidase on the enzymatic conversion of glycan were studied. It was found the alkali-PAA pulp showed excellent enzymatic digestibility. The enzymatic glycan conversion could reach about 80% after 24 h incubation when enzyme loading was 10 FPU/g solid. Simultaneous saccharification and fermentation (SSF) results indicated that the pulp could be well converted to ethanol. Compared with dilute acid pretreated bagasse (DAPB), alkali-PAA pulp could obtain much higher ethanol and xylose concentrations. The fermentation broth still showed some cellulase activity so that the fed pulp could be further converted to sugars and ethanol. After the second batch SSF, the fermentation broth of alkali-PAA pulp still kept about 50% of initial cellulase activity. However, only 21% of initial cellulase activity was kept in the fermentation broth of DAPB. The xylose syrup obtained in SSF of alkali-PAA pulp could be well converted to 2,3-butanediol by Klebsiella pneumoniae CGMCC 1.9131.     

Biobleaching of pulp from agricultural residues with enzymes

Pulp from agricultural residues (wheat straw) was bleached with the DE_PD (chlorine dioxide-extraction with soda and hydrogen peroxide-chlorine dioxide) or P sequence (hydrogen peroxide) after enzymatic pretreatment with cartazyme HS.The enzymatic pretreatment increases the final brightness of the pulp after bleaching with the P and DE_PD sequences (+3.7%) and saves bleaching reagents (from 3.5 to 5.2%); however, it also decreases the pulp yield (from 9.3 to 14.1%) and breaking length (from 20.2 to 13.2%), burst index (from 13.1 to 8.2%) and tear index (from 4.2% to 16.8%) of the paper sheets formed from the pulp.The authors wish to express their gratitude to DGICyT, Spanish Ministry of Education and Science, for financial support granted for the realization of this work as part of Project PB 91-0841.     

A novel in vitro bovine cartilage punch model for assessing the regeneration of focal cartilage defects with biocompatible bacterial nanocellulose

Introduction

Current therapies for articular cartilage defects fail to achieve qualitatively sufficient tissue regeneration, possibly because of a mismatch between the speed of cartilage rebuilding and the resorption of degradable implant polymers. The present study focused on the self-healing capacity of resident cartilage cells in conjunction with cell-free and biocompatible (but non-resorbable) bacterial nanocellulose (BNC). This was tested in a novel in vitro bovine cartilage punch model.

Methods

Standardized bovine cartilage discs with a central defect filled with BNC were cultured for up to eight weeks with/without stimulation with transforming growth factor-β1 (TGF-β1. Cartilage formation and integrity were analyzed by histology, immunohistochemistry and electron microscopy. Content, release and neosynthesis of the matrix molecules proteoglycan/aggrecan, collagen II and collagen I were also quantified. Finally, gene expression of these molecules was profiled in resident chondrocytes and chondrocytes migrated onto the cartilage surface or the implant material.

Results

Non-stimulated and especially TGF-β1-stimulated cartilage discs displayed a preserved structural and functional integrity of the chondrocytes and surrounding matrix, remained vital in long-term culture (eight weeks) without signs of degeneration and showed substantial synthesis of cartilage-specific molecules at the protein and mRNA level. Whereas mobilization of chondrocytes from the matrix onto the surface of cartilage and implant was pivotal for successful seeding of cell-free BNC, chondrocytes did not immigrate into the central BNC area, possibly due to the relatively small diameter of its pores (2 to 5 μm). Chondrocytes on the BNC surface showed signs of successful redifferentiation over time, including increase of aggrecan/collagen type II mRNA, decrease of collagen type I mRNA and initial deposition of proteoglycan and collagen type II in long-term high-density pellet cultures. Although TGF-β1 stimulation showed protective effects on matrix integrity, effects on other parameters were limited.

Conclusions

The present bovine cartilage punch model represents a robust, reproducible and highly suitable tool for the long-term culture of cartilage, maintaining matrix integrity and homoeostasis. As an alternative to animal studies, this model may closely reflect early stages of cartilage regeneration, allowing the evaluation of promising biomaterials with/without chondrogenic factors.     

Improved accessibility and reactivity of dissolving pulp for the viscose process: pretreatment with monocomponent endoglucanase

A high accessibility is an essential prerequisite for a homogeneous substitution of cellulose material. In this study, chemical and enzymatic pretreatments to increase the accessibility of cellulose materials have been investigated. Dissolving pulp has been treated with a monocomponent endoglucanase. Fock's method, a microscale process similar to the viscose process, showed an increase in cellulose yield. Simultaneously, the viscosity decreased. To clarify whether the increase in reactivity was due solely to the decrease in the degree of polymerization, the dissolving pulp was also subjected to acid hydrolysis. At a given viscosity level, the enzymatic pretreated pulp had a higher reactivity than the pulp subjected to acid hydrolysis. To achieve 100% reactivity, according to Fock, the acid-treated pulp showed a lower molecular weight compared to the enzymatic-treated pulp. A monocomponent endoglucanase can thus be used to increase the reactivity and accessibility of dissolving pulp in the viscose process.