Influence of steam and dry heat pretreatment on fibre properties and cellulase degradation of cellulosic fibres

Cellulosic fabric samples of cotton, viscose, lyocell and modal were pretreated with steam and dry heat in the range of 100-190°C. The samples were then treated with a Trichoderma reesei cellulase preparation (total culture filtrate - TC), with mechanical agitation, at a high enzyme dosage of 75% by weight of fabric, for 8 hours. Generally, viscose proved to be easily degradable, followed by cotton and modal. The degree of hydrolysis was the least for Lyocell. Dry heat pretreatments exerted a lower influence on degradation rate than steam pretreatments which showed a distinct maximum at a steam temperature of 130°C. The hydrolysis rate varied strongly depending on treatment conditions and fibre type. Water retention values in treated substrates were changed by up to 20% of initial values.     

Pretreatment of cellulosic wastes to increase enzyme reactivity

The enzymatic hydrolysis of cellulose to glucose is generally a slow reaction. Different pretreatments, such as ball milling to a ?200 mesh or swelling in 1–2% NaOH are reported to increase the reactivity considerably. In this work a fiber fraction from cattle manure was treated in an autoclave for 5–30 min at temperatures ranging from 130–200°C. The reactivity of the cellulose, measured by incubating samples with a commercial cellulase preparation for one hour at 50°C and pH 4.8, was increased by a factor of 4–6 compared to NaOH treatment and 10–12 compared to untreated fiber. The increased reaction rate is probably mostly due to an increase in cellulose availability to enzymatic attack, as structural hemicellulose is hydrolyzed and removed during the treatment. Sugars, produced by hemicellulosis, hydrolysis, will react further to give caramelization products. These side reactions were shown to be suppressed by short treatment times. The treated fiber could support growth of a mixed culture of Trichoderma viride and Candida utilis only after washing, indicating the formation of water soluble inhibitory products during treatment. The treatment with high-temperature steam can probably be used also with other cellulosic materials to increase reactivity. This may be an attractive way to prepare low-valued wastes such as manure fibers, straw, stalks, or corn cobs for fermentation processes to increase the protein content or for use directly as ruminant animal feed.     

Cutinase promotes dry esterification of cotton cellulose

Cutinase from Thermobifida fusca was used to esterify the hydroxyl groups of cellulose with the fatty acids from triolein. Cutinase and triolein were pre‐adsorbed on cotton and the reaction proceeded in a dry state during 48 h at 35°C. The cutinase‐catalyzed esterification of the surface of cotton fabric resulted in the linkage of the oleate groups to the glycoside units of cotton cellulose. The superficial modification was confirmed by performing ATR‐FTIR on treated cotton samples and by MALDI‐TOF analysis of the liquors from the treatment of the esterified cotton with a crude cellulase mixture. Modified cotton fabric also showed a significant increase of hydrophobicity. This work proposes a novel bio‐based approach to obtain hydrophobic cotton. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 32:60–65, 2016     

Advantages of a two-step enzymatic process for cotton–polyester blends

Cotton fabric samples were treated with a formulation of a total crude Trichoderma reesei cellulase in a two-step procedure. In the first step, samples were treated at a low liquor ratio by padding through the enzyme formulation at 21°C and 55°C with a wet pickup of 100% and batched for 12 h. The samples were then treated at a high liquor ratio (1:25) with an identical enzyme formulation at 55°C, with intensive agitation. The pre-treatment influenced the overall weight loss and rate of hydrolysis in samples, and the protein concentration in the liquor of the second step. The overall weight loss was 25–28% (w/w) in the two-step procedure compared to a weight loss of 22% (w/w) in the one-step batch hydrolysis.     

Action of purified Trichoderma reesei cellulases on cotton fibers and yarn

In this work the possibility and potential of treating cotton fibers and yarns instead of fabrics with monocomponent cellulases was investigated. Different pretreatments on fibers were performed and tested in order to improve the accessibility of cotton to enzymatic modification. The enzymatic treatments were evaluated microscopically and by analysing the effects of treated fibers on spinnability, yarn evenness, tenacity and pilling. The accessibility of the cotton fibers for cellulases could be increased by different pretreatments. Steaming of fibers prior to enzymatic treatment was found to be an efficient way to increase hydrolysis levels. Cellulase treatments of carded yarns resulted in modification of yarn properties. Decrease in yarn hairiness was observed and the knitted fabric made of the treated yarn showed a lowered tendency towards pilling. In all cases endoglucanase activity rather than cellobiohydrolase activity was responsible for these modifications.     

Surface activation of dyed fabric for cellulase treatment

Surface activation of fabric made from cellulose fibres, such as viscose, lyocell, modal fibres and cotton, can be achieved by printing of a concentrated NaOH-containing paste. From the concentration of reducing sugars formed in solution, an increase in intensity of the cellulase hydrolysis by a factor of six to eight was observed, which was mainly concentrated at the activated parts of the fabric surface. This method of local activation is of particular interest for modification of materials that have been dyed with special processes to attain an uneven distribution of dyestuff within the yarn cross-section, e.g., indigo ring-dyed denim yarn for jeans production. Fabrics made from regenerated cellulose fibres were used as model substrate to express the effects of surface activation on indigo-dyed material. Wash-down experiments on indigo-dyed denim demonstrated significant colour removal from the activated surface at low overall weight loss of 4-5%. The method is of relevance for a more eco-friendly processing of jeans in the garment industry.     

Comparative Resistance and Resilience of Soil Microbial Communities and Enzyme Activities in Adjacent Native Forest and Agricultural Soils

Degradation of soil properties following deforestation and long-term soil cultivation may lead to decreases in soil microbial diversity and functional stability. In this study, we investigated the differences in the stability (resistance and resilience) of microbial community composition and enzyme activities in adjacent soils under either native tropical forest (FST) or in agricultural cropping use for 14 years (AGR). Mineral soil samples (0 to 5 cm) from both areas were incubated at 40°C, 50°C, 60°C, or 70°C for 15 min in order to successively reduce the microbial biomass. Three and 30 days after the heat shocks, fluorescein diacetate (FDA) hydrolysis, cellulase and laccase activities, and phospholipid-derived fatty acids-based microbial community composition were measured. Microbial biomass was reduced up to 25% in both soils 3 days after the heat shocks. The higher initial values of microbial biomass, enzyme activity, total and particulate soil organic carbon, and aggregate stability in the FST soil coincided with higher enzymatic stability after heat shocks. FDA hydrolysis activity was less affected (more resistance) and cellulase and laccase activities recovered more rapidly (more resilience) in the FST soil relative to the AGR counterpart. In the AGR soil, laccase activity did not show resilience to any heat shock level up to 30 days after the disturbance. Within each soil type, the microbial community composition did not differ between heat shock and control samples at day 3. However, at day 30, FST soil samples treated at 60°C and 70°C contained a microbial community significantly different from the control and with lower biomass regardless of high enzyme resilience. Results of this study show that deforestation followed by long-term cultivation changed microbial community composition and had differential effects on microbial functional stability. Both soils displayed similar resilience to FDA hydrolysis, a composite measure of a broad range of hydrolases, supporting the concept of high functional redundancy in soil microbial communities. In contrast, the resilience of the substrate-specific activities of laccase and cellulase were lower in AGR soils, indicating a less diverse community of microorganisms capable of producing these enzymes and confirming that specific microbial functions are more sensitive measurements for evaluating change in the ecological stability of soils.     

TG/DTG/DTA evaluation of flame retarded cotton fabrics and comparison to cone calorimeter data

Unbleached cotton fabrics (UCF) with 12.5% polypropylene scrim treated with two phosphate-urea based fire-retardant (FR) formulations were evaluated for FR properties using thermogravimetry/differential thermogravimetry/differential thermal analysis (TG/DTG/DTA) method. In addition to testing the two FR-treated unbleached cotton fabrics (CF-FR1 and CF-FR2), bleached cotton fabric (BCF) treated with the two FR formulations (BCF-FR1 and BCF-FR2) was evaluated. Both formulations were washable with add-on of FR chemicals at 18.7% (FR1) or 17.4% (FR2) for UCF and 22.5% (FR1) or 24.9% (FR2) for BCF. The decreasing order of sums at maximal rates of samples degradation in air environment according to DTG method was: BCF (21.40%/min)>UCF (12.91%/min)>BCF-FR2 (12.83%/min)>BCF-FR1 (11.68%/min)>CF-FR2 (10.20%/min)>CF-FR1 (9.73%/min). It indicates that both formulations cause the decrease of thermooxidation of the products at slower rates than the starting material. Several endo- and exothermic peaks observed by DTA in inert and oxidative environment gives additional information about the degradation process. The order of decreasing thermal responses of the studied samples based on sums of DTA peak values of endothermic and exothermic peaks in air environment is: UCF (0.597°C/mg)>BCF (0.120°C/mg)>CF-FR1 (0.089°C/mg)>BCF-FR1 (0.077°C/mg)>CF-FR2 (0.062°C/mg)>BCF-FR2 (0.053°C/mg). This is in agreement with the cone calorimeter results according to which the flammability properties are improving with the decreasing heat release rates or ignition time prolongation in order: UCF>CF-FR1>CF-FR2. The advantage of TG/DTG/DTA method is slower linear heating rate, which allows the more detailed evaluation of the light and flammable cotton fabric.     

Recovery of polyhydroxybutyrate (PHB) from Cupriavidus necator biomass by solvent extraction with 1,2‐propylene carbonate

An integrated procedure for the recovery of polyhydroxybutyrate (PHB) produced by Cupriavidus necator based on the extraction with 1,2‐propylene carbonate was evaluated. The effect of temperature (100–145°C) and contact time (15–45 min), precipitation period, and biomass pretreatments (pH shock and/or thermal treatments) on PHB extraction efficiency and polymer properties was evaluated. The highest yield (95%) and purity (84%) were obtained with the combination of a temperature of 130°C and a contact time of 30 min, with a precipitation period of 48 h. Under these conditions, PHB had a molecular weight of 7.4×10⁵, which was the highest value obtained. Lower values (2.2×10⁵) were obtained for higher temperatures (145°C), while lower temperatures resulted in incomplete extraction yields (45–54%). No further yield improvement was achieved with the pH/heat pretreatments, but the polymer's molecular weight was increased to 1.3×10⁶. The PHB physical properties were not significantly affected by any of the tested procedures, as shown by the narrow ranges obtained for the glass transition temperature (4.8–5.0°C), melting temperature (170.1–180.1°C), melting enthalpy (77.8–88.5 J/g) and crystallinity (55–62%). 1,2‐Propylene carbonate was shown to be an efficient solvent for the extraction of PHB from biomass. The precipitation procedure was found to highly influence the polymer recovery and its molecular weight. Although polymer molecular weight and purity were improved by applying pH/heat pretreatment to the biomass, the procedure involves the use of large amounts of chemicals, which increases the recovery costs and makes the process environmentally unfriendly.     

Efficacy of lipase from Aspergillus niger as an additive in detergent formulations: a statistical approach

The efficacy of lipase from Aspergillus niger MTCC 2594 as an additive in laundry detergent formulations was assessed using response surface methodology (RSM). A five-level four-factorial central composite design was chosen to explain the washing protocol with four critical factors, viz. detergent concentration, lipase concentration, buffer pH and washing temperature. The model suggested that all the factors chosen had a significant impact on oil removal and the optimal conditions for the removal of olive oil from cotton fabric were 1.0% detergent, 75 U of lipase, buffer pH of 9.5 and washing temperature of 25°C. Under optimal conditions, the removal of olive oil from cotton fabric was 33 and 17.1% at 25 and 49°C, respectively, in the presence of lipase over treatment with detergent alone. Hence, lipase from A. niger could be effectively used as an additive in detergent formulation for the removal of triglyceride soil both in cold and warm wash conditions.     

Correlations between the temperature dependence of chlorophyll fluorescence and the fluidity of thylakoid membranes

To monitor changes in membrane fluidity in Arabidopsis leaves and thylakoid membranes, we investigated the temperature dependence of a chlorophyll fluorescence parameter, minimum fluorescence (Fo), and calculated the threshold temperature [T(Fo)] at which the rise of the fluorescence level Fo was considered to be started. For the modification of membrane fluidity we took three different approaches: (1) an examination of wild‐type leaves initially cultured at room temperature (22°C), then exposed to either a lower (4°C) or higher (35°C) temperature for 5 days; (2) measurements of the shift in T(Fo) by two mutants deficient in fatty acid desaturase genes – fad7 and fad7fad8 and (3) an evaluation of the performance of wild‐type plants when leaves were infiltrated with chemicals that modify fluidity. When wild‐type plants were grown at 22°C, the T(Fo) was 48.3 ± 0.3°C. Plants that were then transferred to a chamber set at 4 or 35°C showed a shift in their T(Fo) to 42.7 ± 0.9°C or 48.9 ± 0.1°C, respectively. Under low‐temperature acclimation, the decline in this putative transition temperature was significantly less in fad7 and fad7fad8 mutants compared with the wild‐type. In both leaf and thylakoid samples, values for T(Fo) were reduced in samples treated with benzyl alcohol, a membrane fluidizer, whereas T(Fo) rose in samples treated with dimethylsulfoxide, a membrane rigidifier. These results indicate that the heat‐induced rise of chlorophyll fluorescence is strongly correlated with the fluidity of thylakoid membranes.     

Enzymes and chelating agent in cotton pretreatment

Desized cotton fabric and cotton seed-coat fragments (impurities) have been treated with commercial cellulase (Celluclast 1.5 L), hemicellulase–pectinase (Viscozyme 120 L) and xylanase (Pulpzyme HC) enzymes. Seed-coat fragments hydrolyzed much faster than the cotton fabric itself. This relative difference in hydrolysis rates makes possible a direct enzymatic removal of seed-coat fragments from desized cotton fabric. Addition of chelating agents such as ethylenediamine-tetra-acetic acid (EDTA) markedly enhanced the directed enzyme action. Pretreatments carried out in acidic solution at pH 5 increased the lightness of seed-coat fragments, contrary to the samples treated in neutral medium at pH 7. Alkaline scouring resulted in darker seed-coat fragments except for the samples pretreated with Pulpzyme HC plus EDTA. This effect is similar to that observed in the biobleaching process in pulp and paper industry.     

Enhancing antimicrobial properties of dyed and finished cotton fabrics

1,2,3-Benzothiazole-7-thiocarboxylic acid-S-methylester (commercially known as Actigard^® AM-87) was utilized to impart cotton fabric durable antimicrobial properties. Finishing treatment was carried out under a variety of conditions. The latter were included, effect of pH, concentration of antibacterial agents, curing temperature and curing time. The effect of fabric construction, mercerization, and dyeing with different dyestuff were also investigated. The study was also extended to investigate the technical feasibility of combining antimicrobial finishing treatment in question with other finishing treatment generally carried out on cotton fabric, like soft finishing and crease recovery finishing. The treated fabrics were monitored for antimicrobial properties before and after washing. The treated fabrics were also evaluated for the physio-mechanical properties like fabric tensile strength, elongation at break (or bursting strength for knitted fabric), wettability, crease recovery angle, whiteness index and roughness. Results obtained show that, the most appropriate conditions for treatment cotton fabric with Actigard^® are: padding the cotton fabric in aqueous solution containing 6% Actigard^® at pH 5 (adjusted using formic acid) then squeezed to wet pick up of 100%, dried at 80 °C for 5 min then cured at 100 °C for 150 s. The untreated cotton fabric did not show any antimicrobial activity towards Staphylococcus aureus or Escherichia coli. Treatment of cotton fabric with Actigard^® improves its antimicrobial properties towards S. aureus or E. coli. It is also observed that, treatment of cotton fabric with Actigard^® marginally decreases fabric tensile strength, elongation at break, roughness and WI, whereas; both wettability and crease recovery angle remain practically intact. This was observed whether the fabric was pre-mercerized or not.     

Novel Single-step Pretreatment of Steam Explosion and Choline Chloride to De-lignify Corn Stover for Enhancing Enzymatic Edibility

It is important to develop efficient and economically feasible pretreatment methods for lignocellulosic biomass, to increase annual biomass production. A number of pretreatment methods were introduced to promote subsequent enzymatic hydrolysis of biomass for green energy processes. Pretreatment with steam explosion removes the only xylan at high severity but increases lignin content. In this study, corn stover soaked in choline chloride solution before the steam explosion is economically feasible as it reduced cost. Enzymatic hydrolysis of de-lignified corn stover is enhanced by combinatorial pretreatments of steam explosion and choline chloride. Corn stover pretreated with choline chloride at the ratio of 1:2.2 (w/w), 1.0 MPa, 184 °C, for 15 min efficiently expelled 84.7% lignin and 78.9% xylan. The residual solid comprised of 74.59% glucan and 7.51% xylan was changed to 84.2% glucose and 78.3% xylose with enzyme stacking of 10FPU/g. This single-step pretreatment had ∼ 4.5 and 6.4 times higher glucose yield than SE-pretreated and untreated corn stover, respectively. Furthermore, SEM, XRD and FTIR indicated the porosity, crystalline changes, methoxy bond-cleavage respectively due to the lignin and hemicellulose expulsion. Thus, the released acetic acid during this process introduced this novel strategy, which significantly builds the viability of biomass in short pretreatment time.     

Thermotolerance of IVM‐derived bovine oocytes and embryos after short‐term heat shock

A series of experiments were designed to study the effect of elevated temperatures on developmental competence of bovine oocytes and embryos produced in vitro. In experiment 1, the effect of heat shock (HS) by a mild elevated temperature (40.5°C) for 0, 30, or 60 min on the viability of in vitro matured (IVM) oocytes was tested following in vitro fertilization (IVF) and culture. No significant difference was observed between the control (39°C) and the heat‐treated groups in cleavage, blastocyst formation, or hatching (P > 0.05). In experiment 2, when the HS temperature was increased to 41.5°C, neither the cleavage rate nor blastocyst development was affected by treatment. However, the rate of blastocyst hatching appeared lower in the HS groups (13% in control group vs. 3.9% and 5.6% in 30 min and 60 min, respectively; P < 0.05). When IVM oocytes were treated at 43°C prior to IVF (experiment 3), no difference was detected in blastocyst and expanded blastocyst development following heat treatment for 0, 15, or 30 min, but heat treatment of oocytes for 45 or 60 min significantly reduced blastocyst and expanded blastocyst formation (P < 0.05). In experiment 4, the thermotolerance of day 3 and day 4 bovine IVF embryos were compared. When embryos were pre‐treated with a mild elevated temperature (40.5°C) for 1 hr, and then with a higher temperature (43°C) for 1 hr, no improvement in thermotolerance of the embryos was observed as compared to those treated at 43°C alone. However, a higher thermotolerance was observed in day 4 than day 3 embryos. In conclusion, treatment at 43°C, but not 40.5°C or 41.5°C significantly reduced oocyte developmental competence. An increase in thermotolerance was observed from day 3 to day 4 of in vitro embryonic development, which corresponds to the maternal to zygotic transition of gene expression in bovine embryos. Mol. Reprod. Dev. 53:336–340, 1999. © 1999 Wiley‐Liss, Inc.     

Bioconversion of municipal solid waste to glucose for bio-ethanol production

Selected biodegradable municipal solid waste fractions were subjected to fifteen different pre-hydrolysis treatments to obtain the highest glucose yield for bio-ethanol production. Pre-hydrolysis treatments consisted of dilute acid (H₂SO₄, HNO₃ or HCl, 1 and 4%, 180 min, 60°C), steam treatment (121 and 134°C, 15 min), microwave treatment (700 W, 2 min) or a combination of two of them. Enzymatic hydrolysis was carried out with Trichoderma reesei and Trichoderma viride (10 and 60 FPU g⁻¹ substrate). Glucose yields were compared using a factorial experimental design. The highest glucose yield (72.80%) was obtained with a pre-hydrolysis treatment consisting of H₂SO₄ at 1% concentration, followed by steam treatment at 121°C, and enzymatic hydrolysis with Trichoderma viride at 60 FPU g⁻¹ substrate. The contribution of enzyme loading and acid concentration was significantly higher (49.39 and 47.70%, respectively), than the contribution of temperature during steam treatment (0.13%) to the glucose yield.     

Green bioprocess of degumming of jute fibers and bioscouring of cotton fabric by recombinant pectin methylesterase and pectate lyases from Clostridium thermocellum

Enzymatic processes are emerging as important green biotechnological processes in textile industry. The application of recombinant pectin methylesterase (CtPME) and pectate lyase (CtPL1B) from Clostridium thermocellum for enzymatic degumming of jute or bioscouring of cotton was evaluated. The effectiveness of processes by combination of two enzymes were evaluated that effective degumming of jute and bioscouring of cotton as compared with individual enzyme. The optimum concentrations of two enzymes mixture for both processes, degumming of jute and bio scouring of cotton were 5 mg/mL (2.1 U/mL) of CtPME and 5 mg/mL (3.0 U/mL) of CtPL1B under optimized conditions of 60 min, 100 rpm and 50 °C. FESEM images showed more effective removal of pectin from jute fiber and cotton fabric by enzyme mixture, nevertheless similar to NaOH treatment. Wettability analysis showed mixture of enzymes and NaOH treated cotton fabric absorbed a water drop in 10 s and 8 s, respectively. UTM analysis showed higher tensile strength and Young’s modulus for jute fiber and cotton fabric treated with enzyme mixture than untreated and were similar to those of NaOH treated. These results showed that the CtPME and CtPL1B mixture can be used for replacing the chemical process by green bioprocess in textile industry.     

Implementation of batchwise bioscouring of cotton knits

The examination of critical factors determining the performance of bioscouring showed that a short treatment of the fabric at greater than 80°C after pectinase treatment at 60°C was essential for removal of waxes from the fabric as demonstrated by diminished intensities of methylene peaks in FT-IR measurements. Batch-wise bioscouring of cotton knits was carried out several times with post-treatment at 80°C using a rapid dyeing machine. The dye-ability of bioscoured knits was as good as the company's alkaline scoured ones with slightly higher K/S values. Water pollution caused by effluents of bioscouring and alkaline processes were estimated, as well as that due to the input of chemicals and enzymes. Higher BOD:COD_Cr ratios for enzymes indicated their biodegradable character. After calculation of energy consumption using a simulation program, an economic evaluation of the two processes was done on the basis of one ton production by considering the costs of chemicals and enzyme, water usage, energy consumption and waste water treatment charge.     

The characterization of a pectin-degrading enzyme from Aspergillus oryzae grown on passion fruit peel as the carbon source and the evaluation of its potential for industrial applications

An extracellular pectinase (PEC-I) was isolated from the crude extract of Aspergillus oryzae when grown on passion fruit peel (PFP) as the carbon source and partially purified by ultra filtration, gel filtration and ion-exchange chromatography procedures. Pectinase activity was predominantly found in the retentate. The pectinase from retentate (PEC-Ret) was most active at 50?°C and pH 7.0 and stable at 50?°C with a half-life of approximately 8?h. PEC-I showed higher activity at pH 4.5 and 55?°C, 70?°C and 75?°C and was inhibited by cations (Ag⁺, Fe²⁺, Fe³⁺, Co²⁺, Ca²⁺ and Hg²⁺), EDTA, tannic acid and vanillin. On the other hand, PEC-I was activated by Cu²⁺, ferulic acid, cinnamic acid and 4-hydroxybenzoic acid. The gel under denaturing conditions of PEC-Ret and PEC-I samples showed a protein band of ～45?kDa coincident with that found by staining for pectinase activity. In the bioscouring of cotton fabric the PEC-Ret pectinase preparation led to a better wettability and removed more pectin from the cotton fibers than the commercial enzyme preparation Viscozyme L, but was less effective than a commercial alkaline pectate lyase preparation and alkaline scouring. The incubation of PEC-Ret with guava juice resulted in a 4.15% decrease in juice viscosity.     

The use of steam/air mixtures for partially sterilizing soils infested with cucumber root rot pathogens

An apparatus for experimentally heating glasshouse soils in situ with steam/ air mixtures is described. Covering soil with a rigid hood enabled mixtures to be introduced at ground level at pressures sufficient to give reasonable rates of heat penetration. Heat-treating soils (66–100° C), in which cucumbers had previously grown poorly, increased yields of subsequent crops from (1) 33 to 39/43 kg/plot of four plants and (2) 95 to 178/191 kg/plot. These increases were associated with the eradication of a dark sclerotial pathogenic fungus causing tap root rot. Increasing partial sterilization temperatures increased concentrations of soluble and exchangeable soil Mn from 4 p.p.m. in unheated soil to 9, 43, 87 and 108 p.p.m. at 66, 77, 88 and 100° C respectively, these differences being paralleled by concentrations of Mn in leaf laminae. Cucumbers grown in soils sterilized at 88° C yielded 8 % more than those in soils treated at 100° C.