Enzymatic polymerization on the surface of functionalized cellulose fibers

Enzymatic coating of functionalized cellulose fibers with catechol was performed in the presence of Trametes hirsuta laccase. Cellulose functionalization was done by covalent fixation of aromatic amines onto the cellulose surface using a dyeing procedure with C.I. Reactive Black 5 (RB5) followed by reduction with sodium hydrosulfite. Cellulase enzymes were used on coated and control samples to obtain the analytes linked with the soluble sugars in solution, to prove the reaction concepts described in this paper. Hydrolyzed coated-cellulose showed lower concentration of reducing sugars (1188 mg/L) than control samples (2011 mg/L). The structures of these compounds were checked by LC/MS analysis confirming the presence of functionalized glucose and cellobiose units coupled to poly(catechol) molecules (m/z 580 and m/z 633). Alkali extraction method showed to be very promising to coat cellulose fibers with phenols in the presence of enzymes, at mild conditions of temperature and pH

Bioprocessing of bamboo materials

Bamboo culms were processed using microorganisms, and fibre bundles were obtained. Bacteria and fungi with xylanase activity were isolated from the bamboo retting system. Chemical composition analysis of the fibre bundles obtained showed that the components are mainly cellulose, hemicelluloses and lignin. An increase in cellulose and hemicelluloses content was detected along with a decrease in lignin content after bio-processing. Envi-ronmental Scanning Electronic Microcopy of the fibre bundles (retted) showed great fibre sensibility towards moisture, which could significantly influence mechanical properties. Our results suggested that the bio-processing presented herein contributes to the possible development of a new means of bamboo bio-processing that can be regarded as a primary process to separate fibre bundles from non-cellulosic tissue in the culm.This work was supported by the earmarked fund for Modern Agro-industry Technology Research System (nycytx-19-E23) and the Fundamental Research Funds for the Central Universities JUSRP211A02

Purification and mechanistic characterisation of two polygalacturonases from Sclerotium rolfsii

Sclerotium rolfsii (strain CBS 350.80) was found to produce extraordinary high amounts of polygalacturonases (PGs). Two of these extracellular enzymes were purified by a recently introduced preparative electrophoretic device (isoelectric focusing mode of free flow electrophoresis). PG 1 (39.5 kDa, pI 6.5) and PG 2 (38 kDa, pI 5.4) exhibited quite similar properties, they were found to be both endo-acting enzymes. Both PGs cleaved penta- and trigalacturonic acid while tetragalacturonic acid was only cleaved when trigalacturonic acid was present. The latter substrate was hydrolysed much faster by PG 2. Both enzymes were active on pectins with different degrees of esterification, they were sensitive towards Ca-cations and not glycosylated. The kinetic properties were measured by viscosimetry with polygalacturonic acid as a substrate. NMR experiments on a model substrate revealed an inverting mechanism of carbohydrate hydrolysis for both enzymes

Enzymatic colouration with laccase and peroxidases : recent progress

Enzymes have received significant attention as alternative catalysts to chemical auxiliaries in textile processing. For example, laccases and peroxidises are promising alternatives for bleaching and denim stone washing processes. Similarly, the ability to oxidise different phenolic substrates and dye precusors resulting in the formation of different coloured polymeric molecules is being exploited for developing green chemistry dyeing processes. The enzymatic process is simpler than conventional coloration processes, giving economic and environmental benefits. In this review, the applications of laccase and peroxidise enzymes in dyeing processes of various textile meterials is discussed.This work was supported by the Fundamental Research Funds for the Central Universities JUSRP211A02 and JUSRP21001; the Open Project Program of Key Laboratory of Eco-Textiles, Ministry of Education, Jiangnan University KLET1007 and the Open Project Program of State Key Laboratory for Modification of Chemical Fibers and Polymer Materials, DongHua University LK1005

Tailoring elastase inhibition with synthetic peptides

Chronic wounds are the result of excessive amounts of tissue destructive proteases such as human neutrophil elastase (HNE). The high levels of this enzyme found on those types of wounds inactivate the endogenous inhibitor barrier thus, the search for new HNE inhibitors is required. This work presents two new HNE inhibitor peptides, which were synthesized based on the reactive-site loop of the Bowman–Birk inhibitor protein. The results obtained indicated that these new peptides are competitive inhibitors for HNE and, the inhibitory activity can be modulated by modifications introduced at the N- and C-terminal of the peptides. Furthermore, these peptides were also able to inhibit elastase from a human wound exudate while showing no cytotoxicity against human skin fibroblasts in vitro, greatly supporting their potential application in chronic wound treatment.We would like to acknowledge FCT - Portuguese Foundation for Science and Technology for the scholarship concession; European project Lidwine, contract no. NMP2-CT-2006-026741

Enzymatic synthesis of Tinuvin

Coupling of 3-(3-tert-butyl-4-hydroxyphenyl) propionic acid methylester to 1H-benzotriazole using a laccase from Trametes hirsuta was studied. The potentially resulting coupling product Tinuvin 1130 is an important UV-absorber used in polymer based materials. Oxidation of the phenol by the laccase led to homomolecular coupling reactions while the laccase did not attack 1H-benzotriazole. Due to the homomolecular reaction of the phenol in the presence of laccase coupling of phenol and 1H-benzotriazole was only observed when 1H-benzotriazole was applied in four-fold molar excess. The reaction was monitored by UV/vis spectroscopy, TLC and MS (ion trap) analysis. Coupling of 1H-benzotriazole took place in ortho position according to the postulated mechanism

Polyphenol oxidases exhibit promiscuous proteolytic activity

Tyrosinases are an industrially significant class of polyphenol oxidase. Here, two tyrosinases are shown to cleave a specific peptide bond in a carboxylesterase, yielding a truncated product with higher catalytic activity than the full-length enzyme

Enzymatic reduction and oxidation of fibre-bound azo-dyes

A new customer and environmental friendly method of hair bound dye decolouration was developed. Biotransformation of the azo-dyes Flame Orange and Ruby Red was studied using different oxidoreductases. The pathways of azo dye conversion by these enzymes were investigated and the intermediates and metabolites were identified and characterised using UV–vis spectroscopy, high-performance liquid chromatography (HPLC) and mass spectrometry (MS). Laccase from Pycnoporus cinnabarinus, manganese peroxidase (MnP) from Nematoloma frowardii and the novel Agrocybe aegerita peroxidase (AaP) were found to use a similar mechanism to convert azo dyes. They N-demethylated the dyes and concomitantly polymerized them to some extent. On the other hand the mechanism for cleavage of the azo bond by azo-reductases of Bacillus cereus and B. subtilis was based on reduction of the azo bond at the expense of NAD(P)H

Enzymatic surface hydrolysis of PET enhances bonding in PVC coating

The effect of polyesterase preparations from Thermomyces lanuginosus and Beauveria brongniartii on the hydrophilicity of PET materials was assessed. As a result of polyesterase treatment the hydrophilicity of PET fabrics was increased by up to 8 cm in terms of rising height with increases in surface tension from 6.2 mNm (heat-inactivated control samples) to above 8 mNm. Both enzymes were able to increase the amount of hydroxyl groups on PET from 90 to a maximum of 182 mmol kg-1, while only the B. brongniartii polyesterase released significant amounts of terephthalic acid from PET. Enzymatic surface hydrolysis of PET increased the bonding strength in PVC coating to 13.40 daN 5cm-1 using 0.5% adhesive compared to 11.5 daN 5cm-1 obtained without enzyme pretreatment and 6% of adhesive.The research was financed by the SFG, the FFG, the city of Graz and the province of Styria and by European Commission within the project GRD 2000-30110 Biosynte