Scouring of cotton using marine pectinase

Bioscouring refers to the enzymatic removal of impurities from cotton fibre, which endows it with improved hydrophilicity for further wet processes. In this study, the efficacy of pectinase from newly isolated marine bacteria Bacillus subtilis, isolated from marine sediment; collected from Chinchani beach, Tarapore, India has been evaluated for scouring of cotton fabric and compared with conventional alkaline scouring of cotton. Use of Citrus limetta peel powder as pectin substrate for enzyme production renders pectinase production process more economically viable. Scouring carried out with pectinase dose of 10% (2.8 IU/g of the fabric) on the weight of the fabric at pH 7, 60 °C for 120 min yielded hydrophilic fabric. Physicochemical and mechanical properties of the pectinase scoured fabric were similar to alkaline scoured fabric. Scouring with pectinase preserves fiber's structure and prevents it from deterioration as observed from tensile strength, FTIR and SEM studies against alkaline scoured fabric. Enhanced dye uptake was also observed in case of pectinase scoured cotton fabric as compared to alkaline scoured fabric.     

Degradation kinetics of pectins by an alkaline pectinase in bioscouring of cotton fabrics

Alkaline pectinases have been proven to be effective as bioscouring agents of cotton fabrics. In order to monitor the scouring degree of cotton fabrics quantificationally, a kinetic study of the degradation of pectins in cotton by an alkaline pectinase ‘Bioprep 3000L’ was performed and the influences of initial pectinase concentration and treatment time on bioscouring were evaluated quantitatively. The results showed that although the degradation products increased as pectinase concentration grew higher at same incubation time, the growth multiples of the maximum degradation rate which was used as the starting degradation rate were less than those of initial enzyme concentration. The degradation kinetics of pectins in cotton fibers with a pectinase could be described by modified Ghose–Walseth kinetic empirical equations which had been previously applied to the degradation reaction of cellulose.     

Enzyme processing of textiles in reverse micellar solution

Scouring of cotton using pectinase enzyme, bioscouring, in reverse micellar system was studied. The effectiveness of bioscouring was evaluated by measuring weight loss of cotton, analyzing pectin and cotton wax remaining and by wetness testing. Pectinase enzyme showed excellent activity even in organic media, and the effectiveness of scouring was equivalent or better than that achieved by conventional alkaline process or bioscouring in aqueous media. Enzymatic modification of wool using protease enzyme in the same system was also studied. It has found that felting property and tensile strength of wool fabrics treated by protease in reverse micellar system were superior to those in aqueous media. Possibilities of utilization of the same system for the subsequent textile dyeing process were also investigated. It was found that cotton and polyester fabrics were dyed satisfactorily by reverse micellar system compared to conventional aqueous system.     

Innovative Scouring for Cotton‐Based Textiles

An innovative approach addressing ecological problems associated with scouring of cotton‐based textiles was developed. The innovative scouring method is based on the use of β‐cyclodextrin in the presence of a wetting agent. β‐cyclodextrin is able to accommodate the wax in its cavity, complex with it and dissolve it together with other cotton impurities by the aid of a wetting agent, thereby effecting their removal. The work comprises treatment of desized cotton and polyester/cotton fabrics with β‐cyclodextrin and a wetting agent under a variety of conditions. Variables studied include concentration of β‐cyclodextrin, chemical nature and concentration of the wetting agent, pH of the scouring bath, and temperature and time of scouring. The samples were monitored for the residual wax percent and wettability. The scouring performance of the innovative method was compared with that of the conventional method. Chemical oxygen demand, total dissolved solids and conductivity of the wastewater effluent discharged by the two methods were also determined and compared. The comparison reveals the advantages of the new method in minimizing the degradation of the cotton and polyester/cotton fabrics, which occur during conventional scouring and the persistence of such advantage even after bleaching. The mode of wax removal during the innovative scouring and the impact of the latter on the environment were discussed.     

Analysis of the products from enzymatic scouring of cotton

This article discusses the analysis of the hydrolysis products from one-step scouring of cotton using pectinase and two-step scouring of cotton using lipase then cellulase, protease then cellulase, or lipase/protease then cellulase, to improve water absorbency of cotton. UV spectrophotometric analysis indicated that the pectinase scouring process produced approximately 18-fold higher amounts of reducing sugars and galacturonic acid than any of the two-step scouring processes. The production rate of reducing sugars and galacturonic acid from most of the scouring processes showed a decrease with an increase in time. HPLC analysis revealed that the lipase/protease/cellulase scouring processes produced approximately 5-fold higher amounts of 17 amino acids than the pectinase scouring process. GC analysis for 18 fatty acids (C(8)-C(24)) revealed that three major fatty acids, palmitic acid, stearic acid, and behenic acid, were found on both the scoured and the unscoured fabrics. Scoured fabrics were tested for content of proteins, extractable components, waxes, and anionic components including pectins, and some differences among the fabric scoured with different enzyme combinations were found.     

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.     

New development for combined bioscouring and bleaching of cotton-based fabrics

A thorough investigation into conditions appropriate for effecting combined eco-friendly bioscouring and/or bleaching of cotton-based fabrics was undertaken. Fabrics used include cotton, grey mercerized cotton, cotton/polyester blend 50/50 and cotton/polyester blend 35/65. The four cotton-based fabric were subjected to bioscouring by single use of alkaline pectinase enzymes or by using binary mixtures of alkaline pectinase and cellulase enzymes under a variety of conditions. Results of bioscouring show that, the bioscoured substrates exhibit fabrics performances which are comparable with these of the conventional alkali scouring. It has been also found that, incorporation of ethylenediaminetetraacetic acid (EDTA) in the bioscouring with mixture from alkaline pectinase and cellulase improves the performance of the bioscoured fabrics. Addition of β-cyclodextrin to the bioscouring solution using alkaline pectinase in admixtures with cellulase acts in favor of technical properties and performance of the bioscoured fabrics. Concurrent bioscouring and bleaching by in situ formed peracetic acid using tetraacetylethylenediamine (TAED) and H₂O₂ was also investigated. The results reveal unequivocally that the environmentally sound technology brought about by current development is by far the best. The new development involves a single-stage process for full purification/preparation of cotton textiles. The new development at its optimal comprises treatment of the fabric with an aqueous formulation consisting of alkaline pectinase enzyme (2 g/L), TAED (15 g/L), H₂O₂ (5 g/L), nonionic wetting agent (0.5 g/L) and sodium silicate (2 g/L). The treatment is carried out at 60 °C for 60 min. Beside the advantages of the new development with respect to major technical fabric properties, it is eco-friendly and reproducible. This advocates the new development for mill trials.     

Effect of pectate lyase bioscouring on physical, chemical and low-stress mechanical properties of cotton fabrics

The main objective of the present study was to meticulously investigate an inclusive set of physicochemical and handle properties (determined through Kawabata evaluation system) of bioscoured cotton fabrics. The application of a commercial pectinase preparation, Bioprep 3000L, for a range of concentrations and treatment times, could create a pectin-free textile with low wax content. Multiple regression analysis was used to describe the effect of enzymatic process variables on pectin and waxes removal. Comparison of fabrics' properties such as wettability, whiteness, crystallinity index, and dyeing behaviour, confirmed that bioscouring could be as much effective as the conventional alkaline process. Uncovering the relationship between the composition of materials and their physicochemical properties was attempted. The application of higher enzyme concentrations generated fabrics with improved low-stress mechanical properties. Bending and shear rigidity, compressional resilience, as well as, extensibility of enzymatically treated cotton fabrics could be efficiently predicted by means of a single independent variable, the crystallinity index.     

Validation of leaf and microbial pectinases: commercial launching of a new platform technology

Almost all current genetically modified plant commercial products are derived from seeds. The first protein product made in leaves for commercial use is reported here. Leaf pectinases are validated here with eight liquid commercial microbial enzyme products for textile or juice industry applications. Leaf pectinases are functional in broad pH/temperature ranges as crude leaf extracts, while most commercial enzyme products showed significant loss at alkaline pH or higher temperature, essential for various textile applications. In contrast to commercial liquid enzymes requiring cold storage/transportation, leaf pectinase powder was stored up to 16 months at ambient temperature without loss of enzyme activity. Commercial pectinase products showed much higher enzyme protein PAGE than crude leaf extracts with comparable enzyme activity without protease inhibitors. Natural cotton fibre does not absorb water due to hydrophobic nature of waxes and pectins. After bioscouring with pectinase, measurement of contact‐angle water droplet absorption by the FAMAS videos showed 33 or 63 (leaf pectinase), 61 or 64 (commercial pectinase) milliseconds , well below the 10‐second industry requirements. First marker‐free lettuce plants expressing pectinases were also created by removal of the antibiotic resistance aadA gene. Leaf pectinase powder efficiently clarified orange juice pulp similar to several microbial enzyme products. Commercial pilot scale biomass production of tobacco leaves expressing different pectinases showed that hydroponic growth at Fraunhofer yielded 10 times lower leaf biomass per plant than soil‐grown plants in the greenhouse. Pectinase enzyme yield from the greenhouse plants was double that of Fraunhofer. Thus, this leaf‐production platform offers a novel, low‐cost approach for enzyme production by elimination of fermentation, purification, concentration, formulation and cold chain.     

Pectinolytic activity of bacteria isolated from soil and two fungal strains during submerged fermentation

Seven different strains were selected for their ability to degrade citrus pectin. Alkaline pectinases were produced by five bacterial soil isolates, whereas two fungal strains produced pectinase in an acidic environment. The bacteria were isolated from soil of a plum orchard in Northern Ireland. These isolates produced significant amounts of pectin lyase (PL) and polygalacturonase (PG) with maximum activities of 30.1 and 29.1 U/ml respectively. Fungal strains Aspergillus sp. and PN-1 produced four different pectinolytic activities; endo-PG, exo-PG, pectin esterase (PE) and PL. The Aspergillus sp. produced higher amounts of pectinase than PN-1. The Aspergillus sp. excreted highly stable pectinases, which may be of importance for industrial applications.     

A fast,continuous enzyme-based pretreatment process concept for cotton containing textiles

A fast integrated enzyme-based pretreatment process concept for cotton containing textiles has been developed for operation in the continuous mode. The total processing time for the desizing and scouring operation is 3–10 minutes for fabrics with a weight of 120–300 g/m². Essential elements in the process are the high starting temperature, the presence of surfactant, application of vacuum technology and a robust rinsing process afterwards to remove the degraded pectin together with hydrophobic compounds. This rinsing procedure is realized with water containing chelator and surfactant and at high operation temperature beyond 80°C. A mixture of enzymes is used, consisting of a temperature stable α-amylase and a pectate lyase, both operating under alkaline conditions.     

Pectinase and cellulase activity in the digestive system of the elm leaf beetle,Xanthogaleruca luteola Muller (Coleoptera: Chrysomelidae)

The elm leaf beetle, Xanthogaleruca luteola, is a serious pest of elm trees in urban areas. Partial biochemical characterization of pectinases and cellulases was conducted using the larval digestive system of the pest. Midgut extracts from larvae showed optimum activity for pectinase and cellulase against pectin and carboxymethyl cellulose, respectively, under acidic conditions (pH 6). Pectinases and cellulases were respectively more stable under acidic conditions (pH 4–7) and slightly acidic conditions (pH 5–7) than under highly acidic and alkaline conditions. However, the enzymes were more stable in slightly acidic conditions (pH 6) when incubation time was increased. Maximum activity for the pectinases and cellulases incubated at different temperatures was observed at 45 and 50 °C, respectively. Mg²⁺ remarkably increased pectinase activity, and cellulase activity increased significantly in the presence of Ca²⁺ and Mg²⁺. Sodium dodecyl sulfate significantly decreased pectinase and cellulase activity. The Michaelis–Menten constant (K_M) and the maximal reaction velocity (V_max) values for pectinase were 2 mg·mL^? 1 and 0.017 mmol·min^? 1·mg^? 1 protein toward pectin, respectively. Zymogram analyses revealed the presence of one and five bands of pectinase and cellulase activity, respectively, in the larval midgut extract.     

Characterization of bioscoured cotton fabrics using FT-IR ATR spectroscopy and microscopy techniques

The effects of bioscouring were investigated by characterizing the chemical and physical surface changes of cotton fabrics using a purified pectinase enzyme from Bacillus subtilis strain WSHB04-02. Fourier-transform infrared (FT-IR) attenuated total-reflectance (ATR) spectroscopy, scanning electron microscopy (SEM), and atomic force microscopy (AFM) techniques were employed. FT-IR ATR spectroscopy provided a fast and semi-quantitative assessment of the removal of pectins and/or waxes on the cotton surface by comparing the changes in intensity of the carbonyl peak induced by HCl vapor treatment at around 1736 cm(-1). The bioscoured surface could be clearly distinguished from those of untreated and alkali-treated cotton fibers using a combination of SEM and AFM. The images produced using these techniques revealed that the surface morphography and topography of the cotton fibers were shaped by the etching action mode of pectinases during bioscouring. These findings demonstrated that AFM is a useful supplement to SEM in characterizing cotton surfaces.     

Characterization of cotton fabric scouring by FT-IR ATR spectroscopy

FT-IR attenuated total reflectance (ATR) spectroscopy has been used for the fast characterization of cotton fabric scouring process. The greige and the scoured cotton fabrics showed very similar FT-IR spectrum in transmission mode because the bulk composition of the fabrics are similar. However, FT-IR ATR spectroscopy can provide information about the surface of a fabric. By examination of C–H stretching region at 2800–3000 cm⁻¹, the amount of waxes left on the fabric can be estimated. The presence of pectins and/or waxes can also be probed by observation of carbonyl peak induced by the HCl vapor treatment on the fabric. Based on these changes of FT-IR ATR spectra, the scouring process has been characterized.     

Microbial pectinase: sources, characterization and applications

Today pectinases are upcoming industrially important bacterial enzymes. It can be produced by a variety of microorganisms. These enzymes act on pectin, which is the major component of middle lamella in plant cell wall. Pectinolytic enzymes are classified according to their mode of attack on the galacturonan part of the pectin molecules such as protopectinases, esterase’s and depolymerases. As we know that microbial enzymes work depends up on the type of enzymes application, temperature, concentration, and pH and so on, therefore, pectinase enzyme also differentiated according to their physical and chemical factors too. The biochemical structures of pectinases include members of all the major classes and the structure–function relationship, studies of a few available complexes of pectinases with substrate/analogs could be considered as prototypes for related family member and the molecular characterization of pectinolytic enzymes is also well documented. Furthermore, it provides a bird’s eye view of the possible application of these enzymes in commercial sector.     

Effects of Thermobifida fusca cutinase-carbohydrate-binding module fusion proteins on cotton bioscouring

Previously, we presented a novel approach for increasing Thermobifida fusca cutinase adsorption on cotton fibers by fusing cutinase with a carbohydrate-binding module (CBM). A preliminary study showed that two fusion proteins, namely cutinase-CBM_Cel6A and cutinase-CBM_CenA, with similar stabilities and catalytic properties, had potential applications in bioscouring. In the present study, an indepth analysis of both cutinase-CBMs in bioscouring was explored. Effects of cutinase-CBMs on cotton bioscouring were investigated by characterizing the chemical and physical surface changes in enzyme-treated cotton fabrics. Gas chromatography/mass spectrometry was used to analyze the degradation of the cotton fabric cuticle; Fourier transform infrared microspectroscopy was used to study changes in the chemical composition of the cotton fabric epidermal layer; and scanning electron microscopy was used to monitor minor changes in the morphology of the fiber surface. Our results indicated that cutinase-CBMs in combination with pectinase had a greater effect on cotton fabric than did cutinase. Following scouring with cutinase-CBMs and pectinase, the performance of cotton fabric in terms of its wettability and dyeability was similar to that following alkali scouring. Our study provides a foundation for the further application of cutinase-CBM to bioscouring.     

Ultrasound-boosted enzymatic cotton scouring process with cutinase and pectate lyase

Abstract

The synergistic effect between power ultrasound and enzymes in an enzymatic scouring process has been studied. The scouring enzymes were Fusarium solani pisi cutinase (EC 3.1.1.74) and pectate lyase (EC 4.2.2.2). In different stages of the scouring process, power ultrasound with a pre-optimized power of 0.57 W cm^?2 and a frequency of 30 kHz was applied. It was found that ultrasound shortens the enzymatic scouring process time dramatically; less than 5 min was required to achieve the desired scouring expressed in terms of hydrophilicity of the cotton fiber. The results obtained have been explained in terms of mass transfer intensification by ultrasound (so-called ‘sono-mechanics’) and its effect on the enzyme kinetics (so-called ‘sono-chemistry’). This latter effect has been found by applying ultrasound in a homogeneous enzymatic reaction in which mass transfer did not play any role. The kinetics of product formation in a homogeneous system was carried out using poly-d-galacturonic acid as a model substrate.     

Pectinase Activity Determination: An Early Deceleration in the Release of Reducing Sugars Throws a Spanner in the Works!

Recently, it has been suggested that pectinases could be used to hydrolyze pectin in biorefineries based on pectin-rich agro-industrial wastes. However, for this to be viable, the cost of their production would need to be lowered significantly. In fact, over the last few decades, there have been many attempts to improve pectinase production by existing strains or to screen for new strains from environmental isolates. In these studies, it is necessary to measure pectinase activities. Many researchers use single-time-point assays that involve incubation of pectinolytic extracts with pectic substrates for a fixed time, followed by determination of the liberated reducing sugars. However, different researchers use quite different conditions for this assay. Furthermore, no attention has been given to the reaction profile during the assay. In the current work, we show, for the first time, that a significant deceleration of the rate of liberation of reducing sugars occurs over the first ten minutes of the reaction. As a consequence, the incubation time used in a single-time-point assay has a large effect on the value obtained for the activity. In fact, we demonstrate that, depending on the particular combination of incubation time, pectin concentration and reaction temperature, the same extract could be reported to have activities that differ by an order of magnitude. In addition, we show that the relative activities obtained with polygalacturonic acid do not correlate with those obtained with pectin. We conclude that it is currently impossible to make meaningful comparisons between pectinase activities reported in the literature by workers who have used different assay conditions. Therefore there is an urgent need for the development of a standardized assay for evaluating the saccharification potential of pectinase complexes.