Physicochemical Properties of Cellulose Nanocrystals Extracted from Postconsumer Polyester/Cotton-Blended Fabrics and Their Effects on PVA Composite Films.

The utilisation of cotton waste as precursors in the synthesis of nanocrystalline cellulose has gained significant attention. This approach suggests a sustainable solution to address the growing concern of textile waste accumulation while simultaneously producing a valuable material. The main aim of this study is to examine the properties of cellulose nanocrystals (CNCs) obtained from postconsumer polyester-cotton waste and assess the effect of different fabric structures on the extraction and these properties. To acquire nanocellulose, a thorough decolourisation pretreatment process was utilised, which involved the treatment of polyester-cotton waste with sodium dithionite and hydrogen peroxide. Consequently, the postconsumer material was then treated with an acid hydrolysis method employing a 64% (v/v) sulphuric acid solution at 50 °C for 75 min, resulting in the formation of CNCs with average yield percentages ranging from 38.1% to 69.9%. Separation of the acid from the CNC was facilitated by a centrifugation process followed by dialysis against deionised water. Uniform dispersion was then achieved using ultrasonication. A variety of analytical techniques were employed to investigate the morphological, chemical, thermal, and physical properties of the isolated CNCs. Among these techniques, attenuated total reflection-Fourier-transform infrared spectroscopy (ATR-FTIR), energy-filtered transmission electron microscopy (EF-TEM), thermogravimetric analysis (TGA), and X-ray diffraction (XRD) were utilised to analyse the CNCs. The findings indicated that the separated CNCs exhibited a rod-shaped morphology, measuring between 78 and 358 nm in length and 5 and 16 nm in diameter, and also exhibited high crystallinity (75-89%) and good thermal stability. The extracted CNCs were mixed with polyvinyl alcohol (PVA) and glycerol to assess their reinforcing effect on plastic films. The prepared composite film exhibited improved mechanical properties and thermal stability. Incorporating CNCs led to a 31.9% increase in the tensile strength and a 42.33% rise in the modulus of elasticity. The results from this research proved that CNCs can be extracted from postconsumer mixed fabrics as a potential solution to effectively address the mounting concerns surrounding waste management in the textile industry and also provide avenues for enhancing the qualities of eco-friendly composite films.

Optimization of Xylooligosaccharides Production by Native and Recombinant Xylanase Hydrolysis of Chicken Feed Substrates.

Poultry production faces several challenges, with feed efficiency being the main factor that can be influenced through the use of different nutritional strategies. Xylooligosaccharides (XOS) are functional feed additives that are attracting growing commercial interest due to their excellent ability to modulate the composition of the gut microbiota. The aim of the study was to apply crude and purified fungal xylanases, from Trichoderma harzianum, as well as a recombinant glycoside hydrolase family 10 xylanase, derived from Geobacillus stearothermophilus T6, as additives to locally produced chicken feeds. A Box-Behnken Design (BBD) was used to optimize the reducing sugar yield. Response surface methodology (RSM) revealed that reducing sugars were higher (8.05 mg/mL, 2.81 mg/mL and 2.98 mg/mL) for the starter feed treated with each of the three enzymes compared to the treatment with grower feed (3.11 mg/mL, 2.41 mg/mL and 2.62 mg/mL). The hydrolysis products were analysed by thin-layer chromatography (TLC), and high-performance liquid chromatography (HPLC) analysis and showed that the enzymes hydrolysed the chicken feeds, producing a range of monosaccharides (arabinose, mannose, glucose, and galactose) and XOS, with xylobiose being the predominant XOS. These results show promising data for future applications as additives to poultry feeds.

Enhanced production of a recombinant xylanase (XT6): optimization of production and purification, and scaled-up batch fermentation in a stirred tank bioreactor.

The endoxylanase XT6 produced by Geobacillus stearothermophilus is a desirable candidate for industrial applications. In this study, the gene encoding XT6 was cloned using the pET-28a expression vector and expressed in Escherichia coli BL21 (DE3) cells. Recombinant XT6 production was improved by optimizing cell lysis (sonication, chemical, and enzymatic lysis) and expression conditions. Sonication in a 0.05 M sodium phosphate (pH 6.0) buffer resulted in the highest xylanase activity (16.48 U/ml). Screening and optimization of induction conditions using the Plackett-Burman Design and Box-Behnken Design (BBD) approaches revealed that cell density pre-induction (OD600 nm), post-induction incubation time, and IPTG concentration significantly (p < 0.05) influenced the expression levels of XT6 (16.48 U/ml to 40.06 U/ml) representing a 3.60-fold increase. BBD resulted in a further 8.74-fold increase in activity to 144.02 U/ml. Batch fermentation in a 5-l stirred tank bioreactor at 1 vvm aeration boosted recombinant xylanase production levels to 165 U/ml suggesting that heterologous expression of the XT6 enzyme is suitable for scaled-up production. The pure enzyme with a molecular weight of 43 kDa and a 15.69-fold increase in purity was obtained using affinity chromatography and a cobalt column. Future studies will include application of the purified recombinant xylanase to animal feed.

Screening for cellulases and preliminary optimisation of glucose tolerant ß-glucosidase production and characterisation.

The search for a novel microbial producer of cellulases including a glucose tolerant ß-glucosidase is a challenge as most are inhibited by their product glucose. This study aims to screen for cellulolytic fungi using qualitative and quantitative screening methods. Primary screening revealed 34 of 46 fungal isolates with ß-glucosidase activity. Eleven and 13 of these also displayed endoglucanase and exoglucanase activities, respectively. During secondary screening, this number was reduced to 26 ß-glucosidase producers with 13 also having endoglucanase and exoglucanase activities. Isolate C1 displayed enhanced production of ß-glucosidases in the presence of 0.05 M glucose (69% higher activity). Optimisation of growth conditions for ß-glucosidase production by one variable at a time experiments improved production for (isolates) PS1 (64%), MB5 (84%), and C2 (69%). Isolate PS1 identified as Chaetomella sp. BBA70074 displayed the highest tolerance to glucose, retaining 10% of ß-glucosidase activity in the presence of 0.8 M glucose. Tolerance to glucose increased to 14% when produced under optimal conditions. ß-Glucosidase had a molecular weight of 170 kDa with a pH and temperature optima of 6 and 70°C, respectively. Future studies will include optimisation of the production of the glucose tolerant enzyme by Chaetomella sp. BBA70074.

A glucose tolerant ß-glucosidase from a newly isolated Neofusicoccum parvum strain F7: production, purification, and characterization.

Cellulase-producing microorganisms produce low titres of ß-glucosidases with low tolerance to glucose. This study aimed to improve production, purify, and characterize a ß-glucosidase from a newly isolated Neofusicoccum parvum strain F7. ß-Glucosidase production was significantly enhanced by a sequential statistical modelling approach from 1.5-fold in Plackett-Burman design to 2.5 U/ml in the Box-Behnken design compared to the preliminary one variable at a time experiments (1.6 U/ml). The optimal conditions for enzyme production by BBD were 12 days of fermentation at 20 °C, 175 rpm, 0.5% glycerol and 1.5% casein in pH 6.0 buffer. Three ß-glucosidase isoforms referred to as Bgl1, Bgl2, Bgl3 were purified and characterized from the optimized crude extract displaying IC50 values of 2.6, 22.6 and 319.5 mM for glucose, respectively. Bgl3 with a molecular mass of approximately 65 kDa demonstrated the highest tolerance to glucose among the isoforms. The optimum activity and stability for Bgl3 was observed at pH 4.0 in 50 mM sodium acetate buffer with 80% ß-glucosidase residual activity retained for three hours. This isoform also retained 60% residual activity at 65 °C for one hour which was then reduced to 40% which remained stable for another 90 min. The ß-glucosidase activity of Bgl3 was not enhanced after the addition of metal ions in assay buffers. The Km and vmax for 4-nitrophenyl-ß-D-glucopyranoside were 1.18 mM and 28.08 µmol/min, respectively indicating high affinity for the substrate. The ability to withstand the presence of glucose in conjunction with its thermophilic nature indicates promise for this enzyme in industrial application.

Thermoactive metallo-keratinase from Bacillus sp. NFH5: Characterization, structural elucidation, and potential application as detergent additive.

In recent times, robust green technological developments have advanced the goal of a circular economy by minimizing waste generation. The study was undertaken to explore the keratinolytic activity of chicken feather-degrading bacteria from South African soil. Isolates coded as SSN-01 and HSN-01 were identified as Bacillus sp. NFH5 and Bacillus sp. FHNM and their sequences were deposited in GenBank, with accession numbers MW165830.1 and MW165831.1, respectively. Extracellular enzyme production and thiol group generation by Bacillus sp. NFH5 peaked at 120 h with 1879.09 ± 88.70 U/mL and 9.49 ± 0.78 mM, respectively. Glutamic acid (4.44%), aspartic acid (3.50%), arginine (3.23%), glycine (2.61%), serine (2.08%), and proline (2.08%) were relatively higher in concentration. Keratinase (KerBAN) activity was highest at pH 8.0 and 90 °C but was inhibited by both EDTA and 1,10-phenanthroline. In addition, the keratinase-encoding gene (kerBAN) accessioned OK033360 had 362 amino acid residues, with molecular weight and theoretical isoelectric point of 39 kDa and 8.81, respectively. Findings from this study highlight the significance of Bacillus sp. NFH5 in the bio-recycling of recalcitrant keratinous wastes to protein hydrolysates - potential dietary supplements for livestock feeds. The properties of KerBAN underscore its application potential in green biotechnological processes.

Isolation, screening, preliminary optimisation and characterisation of thermostable xylanase production under submerged fermentation by fungi in Durban, South Africa.

Fungi are renowned for their ability to produce extracellular enzymes into their surrounding environment. Xylanases are hydrolytic enzymes capable of xylan degradation. The objectives of this study were to isolate, screen for potential xylanolytic fungi from soil and tree bark samples from three locations in South Africa and to determine their growth conditions for maximum xylanase production. Forty-six isolates were obtained based on clearing zone formation on xylan-enriched agar plates using Congo red indicator. Xylanase activity was quantified during submerged fermentation. Isolate MS5, identified as Trichoderma harzianum with the highest enzyme activity (38.17 U/ml) was selected for further studies based on thermophilic properties (70°C) and pH (5.0). The culture conditions; incubation period (5 days), agitation speed (160 rpm) wheat bran (1%) and ammonium sulphate (1.2%) were optimised further. Biochemical characterisation of the crude enzyme revealed two pH and temperature optima (pH 6.0 at 60°C and 70°C, pH 8.0 at 55°C and 75°C). The enzyme retained >70% activity after 4 h at pH 6.0 at 70°C. SDS-PAGE revealed multiple protein bands with a prominent band at 70 kDa. Substrate Native PAGE revealed multiple isoforms between 55 and 130 kDa. This enzyme will be beneficial for applications in the animal feed and biofuels industries.

Elucidating the Role of Biofilm-Forming Microbial Communities in Fermentative Biohydrogen Process: An Overview.

Amongst the biofuels described in the literature, biohydrogen has gained heightened attention over the past decade due to its remarkable properties. Biohydrogen is a renewable form of H2 that can be produced under ambient conditions and at a low cost from biomass residues. Innovative approaches are continuously being applied to overcome the low process yields and pave the way for its scalability. Since the process primarily depends on the biohydrogen-producing bacteria, there is a need to acquire in-depth knowledge about the ecology of the various assemblages participating in the process, establishing effective bioaugmentation methods. This work provides an overview of the biofilm-forming communities during H2 production by mixed cultures and the synergistic associations established by certain species during H2 production. The strategies that enhance the growth of biofilms within the H2 reactors are also discussed. A short section is also included, explaining techniques used for examining and studying these biofilm structures. The work concludes with some suggestions that could lead to breakthroughs in this area of research.

Optimization, purification, and characterization of xylanase production by a newly isolated Trichoderma harzianum strain by a two-step statistical experimental design strategy.

Xylanases are hydrolytic enzymes with a wide range of applications in several industries such as biofuels, paper and pulp, food, and feed. The objective of this study was to optimize the culture conditions and medium components for maximal xylanase production from a newly isolated Trichoderma harzianum strain using the Plackett-Burman Design (PBD) and Box Behnken Design (BBD) experimental strategies. Xylanase production was enhanced 4.16-fold to 153.80 U/ml by BBD compared to a preliminary one-factor-at-a-time (OFAT) activity of 37.01 U/ml and 2.24-fold compared to the PBD (68.70 U/ml). The optimal conditions for xylanase production were: 6 days of fermentation, incubation temperature of 70 °C, pH 5.0, agitation of 160 rpm, and 1.2% wheat bran and ammonium sulphate. The experimental design effectively provided conditions for the production of an acidic-thermostable enzyme with exciting potential for application in animal feed improvement. The acidic-thermostable xylanase was purified from the submerged culture and SDS-PAGE analysis revealed a molecular weight of 72 kDa. This protein had maximum xylanolytic activity at pH 6.0 and 65 °C and was stable for 4 h retaining > 70% activity and exhibited substrate specificity for beechwood xylan with a Km of 5.56 mg/ml and Vmax of 1052.63 µmol/min/mg. Enzyme activity was enhanced by Fe2+, Mg2+, and Zn2+. There was an absence of strong inhibitors of xylanase activity. Overall, these characteristics indicate the potential for at least two industrial applications.

Comparative Reinforcement Effect of Achatina fulica Snail Shell Nanoparticles, Montmorillonite, and Kaolinite Nanoclay on the Mechanical and Physical Properties of Greenpoxy Biocomposite.

This study investigated the comparative reinforcement effect of Achatina fulica snail shell nanoparticles, montmorillonite, and kaolinite nanoclay on greenpoxy. Greenpoxy nanocomposites of snail shell nanoparticles, montmorillonite, and kaolinite nanoclay were developed separately, with the nanofiller content ranging from 1 to 3% by weight. Specimens of the nanocomposites with different percentage weights of the nanoparticles were prepared using the resin casting method. Mechanical properties, such as the tensile strength, stiffness, hardness, and impact strength, and water absorption properties of the specimens were evaluated experimentally. It was observed that the incorporation of nanoparticles improved the mechanical properties of pure greenpoxy irrespective of the percentage weight, source, and type of reinforcement. Significantly, the loading of 1 wt.% of snail shell nanoparticles offered superior properties in most cases. Protein fibers and high-concentration calcium carbonate in snail shell nanoparticles, uniform dispersion, and excellent matrix/snail shell nanoparticle adhesion provided a strong structure, resulting in the high strength, stiffness, and decreased water uptake of the composites. The superior properties observed in snail shell nanoparticle composites suggest that this naturally sourced nanofiller can be used as a potential substitute for montmorillonite and kaolinite clays.

Design and Development of Cellulosic Bionanocomposites from Forestry Waste Residues for 3D Printing Applications.

This paper deals with the development of cellulose nanofibres (CNFs) reinforced biopolymers for use in packaging applications. Cellulose nanofibres were extracted from sawdust by a combination of chemical and mechanical treatments. The extracted cellulose nanofibres were chemically modified (fCNFs) and characterised by Fourier Transform Infrared Spectroscopy (FTIR). Bionanocomposites were prepared from biopolymers polylactic acid/polybutylene succinate (PLA/PBS) and cellulose nanofibres by compounding in a twin-screw extruder followed by injection moulding. The developed bionanocomposites were subjected to mechanical and thermal characterisation. As part of product development, CNF-biopolymer pellets were also extruded into filaments which were then 3D printed into prototypes. This work is a successful demonstration of conversion of waste residues into value-added products, which is aligned to the principles of circular economy and sustainable development.

Optimization of cultivation medium and cyclic fed-batch fermentation strategy for enhanced polyhydroxyalkanoate production by Bacillus thuringiensis using a glucose-rich hydrolyzate.

The accumulation of petrochemical plastic waste is detrimental to the environment. Polyhydroxyalkanoates (PHAs) are bacterial-derived polymers utilized for the production of bioplastics. PHA-plastics exhibit mechanical and thermal properties similar to conventional plastics. However, high production cost and obtaining high PHA yield and productivity impedes the widespread use of bioplastics. This study demonstrates the concept of cyclic fed-batch fermentation (CFBF) for enhanced PHA productivity by Bacillus thuringiensis using a glucose-rich hydrolyzate as the sole carbon source. The statistically optimized fermentation conditions used to obtain high cell density biomass (OD600 of 2.4175) were: 8.77 g L-1 yeast extract; 66.63% hydrolyzate (v/v); a fermentation pH of 7.18; and an incubation time of 27.22 h. The CFBF comprised three cycles of 29 h, 52 h, and 65 h, respectively. After the third cyclic event, cell biomass of 20.99 g L-1, PHA concentration of 14.28 g L-1, PHA yield of 68.03%, and PHA productivity of 0.219 g L-1 h-1 was achieved. This cyclic strategy yielded an almost threefold increase in biomass concentration and a fourfold increase in PHA concentration compared with batch fermentation. FTIR spectra of the extracted PHAs display prominent peaks at the wavelengths unique to PHAs. A copolymer was elucidated after the first cyclic event, whereas, after cycles CFBF 2-4, a terpolymer was noted. The PHAs obtained after CFBF cycle 3 have a slightly higher thermal stability compared with commercial PHB. The cyclic events decreased the melting temperature and degree of crystallinity of the PHAs. The approach used in this study demonstrates the possibility of coupling fermentation strategies with hydrolyzate derived from lignocellulosic waste as an alternative feedstock to obtain high cell density biomass and enhanced PHA productivity.

Transformation of pulp and paper mill sludge (PPMS) into a glucose-rich hydrolysate using green chemistry: Assessing pretreatment methods for enhanced hydrolysis.

Pulp and paper mill sludge is a waste stream derived from the pulp and paper making industry, comprised of organic and inorganic material in the form of cellulose, hemicellulose, lignin and ash. In South Africa, approximately fivefour hundred thousand wet tonnes are produced per annum and is currently disposed via landfilling or incineration. However, these disposal methods raise environmental and financial concerns. This waste stream is an attractive feedstock for fermentable sugars, mainly glucose, recovery and can be redirected for valorisation as a feedstock for microbial fermentation to produce value-added products. Sugar recovery by enzymatic hydrolysis, as opposed to acidic hydrolysis, is a promising approach but is hampered by the lignin and inorganic material found in pulp and paper mill sludge. Several treatment steps to reduce or remove these components prior to enzymatic hydrolysis are assessed in this review. Pretreatment improves hydrolysis of cellulosic fibres and ensures a substantial yield of sugars.

Valorisation of waste chicken feathers: Optimisation of keratin extraction from waste chicken feathers by sodium bisulphite, sodium dodecyl sulphate and urea.

Extraction of keratin from keratinous waste materials, such as chicken feathers, has been identified as the favourable approach in beneficiation of this biomass. The chemical extractions of keratin by reducing agents are usually preferred because the process is much faster than its counterpart, oxidation extraction. One such reduction extraction is the use of a mixture of sodium bisulphite, sodium dodecyl sulphate and urea. There are at least five factors that may affect the keratin extraction process and its final properties when using this extraction. Even though this extraction method is often used, the effects of its independent variables have not been studied; as a result, the effects of independent variables cannot be fully linked to the extraction process and final keratin properties. Therefore, this study aimed to optimise the extraction of keratin from waste chicken feathers using sodium bisulphite, sodium dodecyl sulphate and urea. The optimisation was statistically performed using Response Surface Methodology (RSM) linked with Box-Behnken Design. After screening the independent variable using one factor at a time method, the concentration of sodium bisulphite, concentration of sodium dodecyl sulphate, reaction temperature and reaction time were chosen for the study. Twenty-nine experiments were statistically designed and executed, and their results were used to analyse the effects of all the independent variables in order to optimise the extraction process. The reaction temperature was found to be the most significant factor, while the concentration of sodium dodecyl sulphate was the most insignificant factor of this extraction process. Independent variables significance order was reaction temperature > reaction time > concentration of NaHSO3 > concentration of NaC12H25SO4. The designed reduced cubic model was significant and was used to predict the protein yield from the keratin extraction using sodium bisulphite.

Optimisation of surfactant decontamination and pre-treatment of waste chicken feathers by using response surface methodology.

Commercially processed, untreated chicken feathers are biologically hazardous due to the presence of blood-borne pathogens. Prior to valorisation, it is crucial that they are decontaminated to remove the microbial contamination. The present study focuses on evaluating the best technologies to decontaminate and pre-treat chicken feathers in order to make them suitable for valorisation. Waste chicken feathers were washed with three surfactants (sodium dodecyl sulphate) dimethyl dioctadecyl ammonium chloride, and polyoxyethylene (40) stearate) using statistically designed experiments. Process conditions were optimised using response surface methodology with a Box-Behnken experimental design. The data were compared with decontamination using an autoclave. Under optimised conditions, the microbial counts of the decontaminated and pre-treated chicken feathers were significantly reduced making them safe for handling and use for valorisation applications.

Valorisation of chicken feathers: Characterisation of chemical properties.

The characterisation of the chemical properties of the whole chicken feather and its fractions (barb and rachis), was undertaken to identify opportunities for valorizing this waste product. The authors have described the physical, morphological, mechanical, electrical and thermal properties of the chicken feathers and related them to potential valorisation routes of the waste. However, identification of their chemical properties is necessary to complete a comprehensive description of chicken feather fractions. Hence, the chicken feathers were thoroughly characterised by proximate and ultimate analyses, elemental composition, spectroscopic analyses, durability in different solvents, burning test, and hydrophobicity. The proximate analysis of chicken feathers revealed the following compositions: crude lipid (0.83%), crude fibre (2.15%), crude protein (82.36%), ash (1.49%), NFE (1.02%) and moisture content (12.33%) whereas the ultimate analyses showed: carbon (64.47%), nitrogen (10.41%), oxygen (22.34%), and sulphur (2.64%). FTIR analysis revealed that the chicken feather fractions contain amide and carboxylic groups indicative of proteinious functional groups; XRD showed a crystallinity index of 22. Durability and burning tests confirmed that feathers behaved similarly to animal fibre. This reveals that chicken feather can be a valuable raw material in textile, plastic, cosmetics, pharmaceuticals, biomedical and bioenergy industries.