The essential role of architectural noncoding RNA Neat1 in cold-induced beige adipocyte differentiation in mice.

Neat1 is an architectural RNA that provides the structural basis for nuclear bodies known as paraspeckles. Although the assembly processes by which Neat1 organizes paraspeckle components are well-documented, the physiological functions of Neat1 are not yet fully understood. This is partly because Neat1 knockout (KO) mice, lacking paraspeckles, do not exhibit overt phenotypes under normal laboratory conditions. During our search for conditions that elicit clear phenotypes in Neat1 KO mice, we discovered that the differentiation of beige adipocytes-inducible thermogenic cells that emerge upon cold exposure-is severely impaired in these mutant mice. Neat1_2, the architectural isoform of Neat1, is transiently upregulated during the early stages of beige adipocyte differentiation, coinciding with increased paraspeckle formation. Genes with altered expression during beige adipocyte differentiation typically cluster at specific chromosomal locations, some of which move closer to paraspeckles upon cold exposure. These observations suggest that paraspeckles might coordinate the regulation of these gene clusters by controlling the activity of certain transcriptional condensates that coregulate multiple genes. We propose that our findings highlight a potential role for Neat1 and paraspeckles in modulating chromosomal organization and gene expression, potentially crucial processes for the differentiation of beige adipocytes.

Interspecies interactions determine growth dynamics of biopolymer-degrading populations in microbial communities.

Microbial communities perform essential ecosystem functions such as the remineralization of organic carbon that exists as biopolymers. The first step in mineralization is performed by biopolymer degraders, which harbor enzymes that can break down polymers into constituent oligo- or monomeric forms. The released nutrients not only allow degraders to grow, but also promote growth of cells that either consume the degradation products, i.e., exploiters, or consume metabolites released by the degraders or exploiters, i.e., scavengers. It is currently not clear how such remineralizing communities assemble at the microscale-how interactions between the different guilds influence their growth and spatial distribution, and hence the development and dynamics of the community. Here, we address this knowledge gap by studying marine microbial communities that grow on the abundant marine biopolymer alginate. We used batch growth assays and microfluidics coupled to time-lapse microscopy to quantitatively investigate growth and spatial distribution of single cells. We found that the presence of exploiters or scavengers alters the spatial distribution of degrader cells. In general, exploiters and scavengers-which we collectively refer to as cross-feeder cells-slowed down the growth of degrader cells. In addition, coexistence with cross-feeders altered the production of the extracellular enzymes that break down polymers by degrader cells. Our findings reveal that ecological interactions by nondegrading community members have a profound impact on the functions of microbial communities that remineralize carbon biopolymers in nature.

Local response and emerging nonlinear elastic length scale in biopolymer matrices.

Nonlinear stiffening is a ubiquitous property of major types of biopolymers that make up the extracellular matrices (ECM) including collagen, fibrin, and basement membrane. Within the ECM, many types of cells such as fibroblasts and cancer cells have a spindle-like shape that acts like two equal and opposite force monopoles, which anisotropically stretch their surroundings and locally stiffen the matrix. Here, we first use optical tweezers to study the nonlinear force-displacement response to localized monopole forces. We then propose an effective-probe scaling argument that a local point force application can induce a stiffened region in the matrix, which can be characterized by a nonlinear length scale R* that increases with the increasing force magnitude; the local nonlinear force-displacement response is a result of the nonlinear growth of this effective probe that linearly deforms an increasing portion of the surrounding matrix. Furthermore, we show that this emerging nonlinear length scale R* can be observed around living cells and can be perturbed by varying matrix concentration or inhibiting cell contractility.

Interfacial Dehydration Strategy for Chitosan Film Shape Morphing and Its Application.

Shape morphing of biopolymer materials, such as chitosan (CS) films, has great potential for applications in many fields. Traditionally, their responsive behavior has been induced by the differential water swelling through the preparation of multicomponent composites or cross-linking as deformation is not controllable in the absence of these processes. Here, we report an interfacial dehydration strategy to trigger the shape morphing of the monocomponent CS film without cross-linking. The release of water molecules is achieved by spraying the surface with a NaOH solution or organic solvents, which results in the interfacial shrinkage and deformation of the entire film. On the basis of this strategy, a range of CS actuators were developed, such as soft grippers, joint actuators, and a light switch. Combined with the geometry effect, edited deformation was also achieved from the planar CS film. This shape-morphing strategy is expected to enable the application of more biopolymers in a wide range of fields.

An exopolysaccharide pathway from a freshwater Sphingomonas isolate.

Bacteria embellish their cell envelopes with a variety of specialized polysaccharides. Biosynthesis pathways for these glycans are complex, and final products vary greatly in their chemical structures, physical properties, and biological activities. This tremendous diversity comes from the ability to arrange complex pools of monosaccharide building blocks into polymers with many possible linkage configurations. Due to the complex chemistry of bacterial glycans, very few biosynthetic pathways have been defined in detail. As part of an initiative to characterize novel polysaccharide biosynthesis enzymes, we isolated a bacterium from Lake Michigan called Sphingomonas sp. LM7 that is proficient in exopolysaccharide (EPS) production. We identified genes that contribute to EPS biosynthesis in LM7 by screening a transposon mutant library for colonies displaying altered colony morphology. A gene cluster was identified that appears to encode a complete wzy/wzx-dependent polysaccharide assembly pathway. Deleting individual genes in this cluster caused a non-mucoid phenotype and a corresponding loss of EPS secretion, confirming the role of this gene cluster in polysaccharide production. We extracted EPS from LM7 cultures and determined that it contains a linear chain of 3- and 4-linked glucose, galactose, and glucuronic acid residues. Finally, we show that the EPS pathway in Sphingomonas sp. LM7 diverges from that of sphingan-family EPSs and adhesive polysaccharides such as the holdfast that are present in other Alphaproteobacteria. Our approach of characterizing complete biosynthetic pathways holds promise for engineering polysaccharides with valuable properties. IMPORTANCE: Bacteria produce complex polysaccharides that serve a range of biological functions. These polymers often have properties that make them attractive for industrial applications, but they remain woefully underutilized. In this work, we studied a novel polysaccharide called promonan that is produced by Sphingomonas sp. LM7, a bacterium we isolated from Lake Michigan. We extracted promonan from LM7 cultures and identified which sugars are present in the polymer. We also identified the genes responsible for polysaccharide production. Comparing the promonan genes to those of other bacteria showed that promonan is distinct from previously characterized polysaccharides. We conclude by discussing how the promonan pathway could be used to produce new polysaccharides through genetic engineering.

Recent Advances of Bio-Based Hydrogel Derived Interfacial Evaporator for Sustainable Water and Collaborative Energy Storage Applications.

Solar interfacial evaporation strategy (SIES) has shown great potential to deal with water scarcity and energy crisis. Biobased hydrogel derived interfacial evaporator can realize efficient evaporation due to the unique structure- properties relationship. As such, increasing studies have focused on water treatment or even potential accompanying advanced energy storage applications with respect of efficiency and mechanism of bio-based hydrogel derived interfacial evaporation from microscale to molecular scale. In this review, the interrelationship between efficient interfacial evaporator and bio-based hydrogel is first presented. Then, special attention is paid on the inherent molecular characteristics of the biopolymer related to the up-to-date studies of promising biopolymers derived interfacial evaporator with the objective to showcase the unique superiority of biopolymer. In addition, the applications of the bio-based hydrogels are highlighted concerning the aspects including water desalination, water decontamination atmospheric water harvesting, energy storage and conversion. Finally, the challenges and future perspectives are given to unveil the bottleneck of the biobased hydrogel derived SIES in sustainable water and other energy storage applications.

Conformational Selection in Enzyme-Catalyzed Depolymerization of Bio-based Polyesters.

Enzymatic degradation of polymers holds promise for advancing towards a bio-based economy. However, bulky polymers presents challenges in accessibility for biocatalysts, hindering depolymerization reactions. Beyond the impact of crystallinity, polymer chains can reside in different conformations affecting binding efficiency to the enzyme. We previously showed that the gauche and trans chain conformers associated with crystalline and amorphous regions of the synthetic polyethylene terephthalate (PET) display different affinity to PETase, thus affecting the depolymerization rate. However, structural-function relationships for biopolymers remain poorly understood in biocatalysis. In this study, we explored biodegradation of by-us previously synthesized bio-polyesters made from a rigid bicyclic chiral terpene-based diol and copolymerized with various renewable diesters. Herein, four of those polyesters spanning from semi-aromatic to aliphatic were subjected to enzymatic degradations in concert with induced-fit docking (IFD) analyses. Our findings demonstrate the importance of conformational selection in enzymatic depolymerization of biopolymers. A straight or twisted conformation of the polymer chain is crucial in biocatalytic degradation by showing different affinities to enzyme ground-state conformers. This work highlights the importance of considering the conformational match between the polymer and the enzyme to optimize the biocatalytic degradation efficiency of biopolymers, providing valuable insights for the development of sustainable bioprocesses.

Microbial alchemy: upcycling of brewery spent grains into high-value products through fermentation.

Spent grains are one of the lignocellulosic biomasses available in abundance, discarded by breweries as waste. The brewing process generates around 25-30% of waste in different forms and spent grains alone account for 80-85% of that waste, resulting in a significant global waste volume. Despite containing essential nutrients, i.e., carbohydrates, fibers, proteins, fatty acids, lipids, minerals, and vitamins, efficient and economically viable valorization of these grains is lacking. Microbial fermentation enables the valorization of spent grain biomass into numerous commercially valuable products used in energy, food, healthcare, and biomaterials. However, the process still needs more investigation to overcome challenges, such as transportation, cost-effective pretreatment, and fermentation strategy. to lower the product cost and to achieve market feasibility and customer affordability. This review summarizes the potential of spent grains valorization via microbial fermentation and associated challenges.

Starch-chitosan blends: A comprehensive review on the preparation, physicochemical properties and applications.

Starch and chitosan, polysaccharides derived from natural sources, have significant potential across various domains. Starch is extracted from starch-bearing plants, such as potatoes, whereas chitosan is obtained from the exoskeletons of marine animals, fungi and insects. However, the original forms of starch and chitosan have several limitations, such as low solubility and weak mechanical strength. Interestingly, the combined effects of starch and chitosan resulted in the development of starch-chitosan blends with markedly improved functional properties. These blends demonstrated high tensile strength, improved hydrophilicity and increased adsorption capacity. Furthermore, modification of starch-chitosan blends by techniques such as crosslinking and incorporation of other functional materials contributes to diverse characteristics and functionalities. This review addresses a crucial gap in the literature by providing an overview and up-to-date analysis of starch-chitosan blends. The preparation methods and functional properties of these blends in various forms, such as films, beads and hydrogels, have been extensively discussed. Emphasis is placed on the versatile applications of these blends in research, development and industries such as pharmaceuticals, wastewater treatment, agriculture and food technology. This review aims to provide an insightful overview of starch-chitosan blends and stimulate broader interdisciplinary research interests. By providing concluding insights and prospects, this review highlights the potential for further exploration of the impact of starch-chitosan blends on consumers and the environment.

Impact and torsional behavior of additive layer-manufactured biopolymer: An advancement for orthopedic applications.

Distal ulna locking bone plates (DLBPs) are commonly employed in the treatment of distal ulna fractures. However, commercially available metallic bone plates experience stress shielding and lack corrosion resistance. Poly lactic acid (PLA) is highly favored biopolymer due to its biocompatible and bioabsorbable nature with human tissues. The use of additive layer manufacturing (ALM) is gaining attention for creating customized implants with intricate structures tailored to patient autonomy. ALM-based PLA bone plates must provide high resistance against impact and torsional forces, necessitating the adjustment of printing process parameters. This study focuses on examining the influence of key printing parameters, on the impact strength and torque-withstanding capability of DLBPs. Experimental results, along with microscopic images, reveal that an increase in infill density (IF) and wall thickness imparts strong resistance to layers against crack propagation under impact and torsional loads. On the contrary, an increase in layer height and printing speed leads to delamination and early fracture of layers during impact and torsional testing. IF significantly contributes to improving the impact strength and torque-withstanding capability of DLBPs by 70.53% and 80.65%, respectively. The study highlights the potential of the ALM technique in developing DLBPs with sufficient mechanical strength for biomedical applications.

Protein-based microneedles for biomedical applications: A systematic review.

Microneedles are minimally-invasive devices with the unique capability of bypassing physiological barriers. Hence, they are widely used for different applications from drug/vaccine delivery to diagnosis and cosmetic fields. Recently, natural biopolymers (particularly carbohydrates and proteins) have garnered attention as safe and biocompatible materials with tailorable features for microneedle construction. Several review articles have dealt with carbohydrate-based microneedles. This review aims to highlight the less-noticed role of proteins through a systematic search strategy based on the PRISMA guideline from international databases of PubMed, Science Direct, Scopus, and Google Scholar. Original English articles with the keyword "microneedle(s)" in their titles along with at least one of the keywords "biopolymers, silk, gelatin, collagen, zein, keratin, fish-scale, mussel, and suckerin" were collected and those in which the proteins undertook a structural role were screened. Then, we focused on the structures and applications of protein-based microneedles. Also, the unique features of some protein biopolymers that make them ideal for microneedle construction (e.g., excellent mechanical strength, self-adhesion, and self-assembly), as well as the challenges associated with them were reviewed. Altogether, the proteins identified so far seem not only promising for the fabrication of "better" microneedles in the future but also inspiring for designing biomimetic structural biopolymers with ideal characteristics.

Antilisterial and antioxidant exopolysaccharide from Enterococcus faecium PCH.25 isolated from cow butter: characterization and probiotic potential.

Exopolysaccharides produced by lactic acid bacteria have gained attention for their potential health benefits and applications in functional foods. This study explores the isolation and characterization of a novel exopolysaccharide-producing strain from dairy products. The aim was to evaluate its probiotic potential and investigate the properties of the produced exopolysaccharide. A strain identified as Enterococcus faecium PCH.25, isolated from cow butter, demonstrated exopolysaccharide production. The study's novelty lies in the comprehensive characterization of this strain and its exopolysaccharide, revealing unique properties with potential applications in food, cosmetic, and pharmaceutical industries. The E. faecium PCH.25 strain exhibited strong acid tolerance, with a 92.24% viability rate at pH 2 after 2 h of incubation. It also demonstrated notable auto-aggregation (85.27% after 24 h) and co-aggregation abilities, antibiotic sensitivity, and absence of hemolytic activity, suggesting its probiotic potential. The exopolysaccharide produced by this strain showed bactericidal activity (MIC and MBC = 1.8 mg/ml) against Listeria monocytogenes and antioxidant properties (22.8%). Chemical analysis revealed a heteropolysaccharide composed of glucose and fructose monomers, with various functional groups contributing to its bioactivities. Physical characterization of the exopolysaccharide indicated thermal stability up to 270 °C, a negative zeta-potential (-27 mV), and an average particle size of 235 nm. Scanning electron microscopy and energy dispersive X-ray analysis revealed a smooth, nonporous structure primarily composed of carbon and oxygen, with an amorphous nature. These findings suggest that the exopolysaccharide from E. faecium PCH.25 has potential as a natural antibacterial and antioxidant polymer for use in functional foods, cosmetics, and pharmaceuticals.

Starch/chitosan nanoparticles bionanocomposite membranes for methylene blue dye removal.

This research aims to develop relatively new membranes from starch biopolymer incorporated with different concentrations (0, 5, 10, 15, 20% w/w of solid starch) of chitosan nanoparticles (CNP) that can be used for water treatment. The membranes were fabricated using the solvent casting method while the CNP was produced using the ionic gelation method. The membranes were characterized in terms of morphology, porosity, water vapor permeability (WVP), and water contact angle. The application of the membranes to treat water was demonstrated on methylene blue solution because methylene blue is a commonly used dye in many industries. It was found that the starch/10% CNP membrane was the optimum membrane for methylene blue dye treatment because the membrane exhibits a smooth surface, high WVP (1.67 × 10-10g Pa-1h-1m-1), high porosity (59.92%), low water contact angle value (44.8°), and resulted in the highest percentage removal of methylene blue (94.0%) after the filtration. After filtration, the starch/10% CNP membrane was still in good condition without breakage. In conclusion, the starch/CNP membranes produced in this study are promising for sustainable and environmentally friendly water treatment, especially for water containing methylene blue dye. This research aligns with current thematic trends in bionanohybrid composite materials utilization, offering innovative solutions for addressing water pollution challenges.

Intensive Microalgal Cultivation and Tertiary Phosphorus Recovery from Wastewaters via the EcoRecover Process.

Mixed community microalgal wastewater treatment technologies have the potential to advance the limits of technology for biological nutrient recovery while producing a renewable carbon feedstock, but a deeper understanding of their performance is required for system optimization and control. In this study, we characterized the performance of a 568 m3·day-1 Clearas EcoRecover system for tertiary phosphorus removal (and recovery as biomass) at an operating water resource recovery facility (WRRF). The process consists of a (dark) mix tank, photobioreactors (PBRs), and a membrane tank with ultrafiltration membranes for the separation of hydraulic and solids residence times. Through continuous online monitoring, long-term on-site monitoring, and on-site batch experiments, we demonstrate (i) the importance of carbohydrate storage in PBRs to support phosphorus uptake under dark conditions in the mix tank and (ii) the potential for polyphosphate accumulation in the mixed algal communities. Over a 3-month winter period with limited outside influences (e.g., no major upstream process changes), the effluent total phosphorus (TP) concentration was 0.03 ± 0.03 mg-P·L-1 (0.01 ± 0.02 mg-P·L-1 orthophosphate). Core microbial community taxa included Chlorella spp., Scenedesmus spp., and Monoraphidium spp., and key indicators of stable performance included near-neutral pH, sufficient alkalinity, and a diel rhythm in dissolved oxygen.

The Influence of Various Crosslinking Conditions of EDC/NHS on the Properties of Fish Collagen Film.

The process of crosslinking improves the physicochemical properties of biopolymer-based composites, making them valuable for biomedical applications. EDC/NHS-crosslinked collagen materials have a significant potential for tissue engineering applications, due to their enhanced properties and biocompatibility. Chemical crosslinking of samples can be carried out in several ways, which is crucial and has a direct effect on the final properties of the obtained material. In this study, the effect of crosslinking conditions on the properties of collagen films using EDC and NHS was investigated. Studies included FTIR spectroscopy, AFM, swelling and degradation tests, mechanical testing and contact angle measurements. Evaluation of prepared collagen films indicated that both crosslinking agents and crosslinking conditions influenced film properties. Notable alternations were observed in the infrared spectrum of the sample, to which EDC was added directly to the fish collagen solution. The same sample indicated the lowest Young modulus, tensile strength and breaking force parameters and the highest elongation at break. All samples reached the maximum swelling degree two hours after immersion in PBS solution; however, the immersion-crosslinked samples exhibited a significantly lower degree of swelling and were highly durable. The highest roughness was observed for the collagen film crosslinked with EDC, whereas the lowest was observed for the specimen crosslinked with EDC with NHS addition. The crosslinking agents increased the surface roughness of the collagen film, except for the sample modified with the addition of EDC and NHS mixture. All films were characterized by hydrophilic character. The films' modification resulted in a decrease in their hydrophilicity and wettability. Our research allows for a comparison of proposed EDC/NHS crosslinking conditions and their influence on the physicochemical properties of fish collagen thin films. EDC and NHS are promising crosslinking agents for the modification of fish collagen used in biomedical applications.

Chitosan-Based Oleogels: Emulsion Drying Kinetics and Physical, Rheological, and Textural Characteristics of Olive Oil Oleogels.

Oleogels are of high interest as promising substitutes for trans fats in foods. An emulsion-templated method was used to trap olive oil in the chitosan crosslinked with vanillin matrix. Oil in water emulsions (50:50 w/w) with different chitosan content (0.7 and 0.8% w/w) with a constant vanillin/chitosan ratio (1.3) were air-dried at different temperatures (50, 60, 70, and 80 °C) and freeze-dried (-26 °C and 0.1 mbar) to produce oleogels. Only falling rate periods were determined during air-drying kinetics and were successfully modeled with empirical and diffusional models. At a drying temperature of 70 °C, the drying kinetics were the fastest. The viscoelasticity of oleogels showed that the elastic modulus significantly increased after drying at 60 and 70 °C, and those dried at 50 °C and freeze-dried were weaker. All oleogels showed high oil binding capacity (>91%), but the highest values (>97%) were obtained in oleogels with a threshold elastic modulus (50,000 Pa). The oleogels' color depended on the drying temperature and chitosan content (independent of the drying method). Significant differences were observed between air-dried and freeze-dried oleogels with respect to oxidative stability. Oxidation increased with the air-drying time regardless of chitosan content. The found results indicated that drying conditions must be carefully selected to produce oleogels with specific features.

Three-dimensional label-free visualization and quantification of polyhydroxyalkanoates in individual bacterial cell in its native state.

Polyhydroxyalkanoates (PHAs) are biodegradable polyesters that are intracellularly accumulated as distinct insoluble granules by various microorganisms. PHAs have attracted much attention as sustainable substitutes for petroleum-based plastics. However, the formation of PHA granules and their characteristics, such as localization, volume, weight, and density of granules, in an individual live bacterial cell are not well understood. Here, we report the results of three-dimensional (3D) quantitative label-free analysis of PHA granules in individual live bacterial cells through measuring the refractive index distributions by optical diffraction tomography (ODT). The formation and growth of PHA granules in the cells of Cupriavidus necator, the best-studied native PHA producer, and recombinant Escherichia coli harboring C. necator poly(3-hydroxybutyrate) (PHB) biosynthesis pathway are comparatively examined. Through the statistical ODT analyses of the bacterial cells, the distinctive characteristics for density and localization of PHB granules in vivo could be observed. The PHB granules in recombinant E. coli show higher density and localization polarity compared with those of C. necator, indicating that polymer chains are more densely packed and granules tend to be located at the cell poles, respectively. The cells were investigated in more detail through real-time 3D analyses, showing how differently PHA granules are processed in relation to the cell division process in native and nonnative PHA-producing strains. We also show that PHA granule-associated protein PhaM of C. necator plays a key role in making these differences between C. necator and recombinant E. coli strains. This study provides spatiotemporal insights into PHA accumulation inside the native and recombinant bacterial cells.

Effectiveness of chitosan nanohydrogel mediated encapsulation of EcR dsRNA against the whitefly, Bemisia tabaci Asia-I (Gennedius) (Hemiptera: Aleyordidae).

Bemisia tabaci is a global invasive pest causing substantial loss on several economically important crops and has developed a very high level of resistance to insecticides making current management practices ineffective. Thus, the novel pest management strategy like RNA interference (RNAi) has emerged as a potential molecular tool in the management of insect pests particularly B. tabaci. The present study investigated RNAi mediated silencing of the Ecdysone Receptor (EcR) gene in B. tabaci Asia-I using biodegradable Chitosan Nanoparticles (CNPs) hydrogel containing EcR dsRNA. The formation of nanohydrogel and dsRNA loading were characterized by gel retardation assay, scanning electron microscopy (SEM); transmission electron microscopy (TEM) and Fourier transform infrared microscopy (FTIR). The stability of CNPs/dsRNA was assessed by exposure to direct sunlight and UV light for different time periods. The CNPs/dsRNA exhibited increased stability over the untreated control and further confirmed by bioassay studies which yielded mortality over 80% and effectively down regulated the expression of the EcR gene as confirmed by qRT-PCR analysis. These investigations provide potential avenues for advancing innovative pest management strategies using biopolymer CNPs hydrogel, which can enhance the efficiency of dsRNA as a safe and targeted solution in the management of whiteflies.

Synthesis of biopolymer blends nanocomposites embedded with mono-(Ag, Fe) and bi-(Ag-Fe) metallic nanoparticles using an eco-friendly approach for antimicrobial activities.

Plant-mediated solution casting is used to develop eco-friendly polymer blend nanocomposites from polyvinyl alcohol (PVA) and polyvinylpyrrolidone (PVP) doped with Silver (Ag), Ferrous (Fe) monometallic and Silver-Ferrous (Ag-Fe) bimetallic nanoparticles (NPs). These nanocomposites were studied to understand their electromagnetic interface (EMI) shielding efficiency and antimicrobial activities, besides evaluating their physical and chemical properties. The Fourier transform infrared (FTIR), X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), and energy dispersive X-ray (EDX) characterization techniques were used to examine the interactions between the polymers, the presence of silver and ferrous particles in the composites, the crystallinity shift, the surface morphology, the shape and size of the nanoparticles and the distribution of the nanoparticles in the composites. The FTIR spectra showed the interactions among the components of the composites. According to XRD spectra, the incorporation of nanoparticles into the PVA polymer significantly reduced the crystalline character of the polymer from 0.38 to 0.24 for the composition consisting of silver and iron nanoparticles in equal proportion. The results from SEM, EDX and XRD corroborate the presence of nanoparticle forms. The thermogravimetric analysis (TGA) tests reveal that the thermal stability of bimetallic composites is greater than that of monometallic composites. The tensile properties showed that the addition of nanoparticles to the PVA/PVP polymer matrix increased its mechanical strength from 59.3 MPa to 85.5 MPa. We examined its efficacy against Escherichia coli, Staphylococcus aureus and Candida albicans as microorganisms. Good antibacterial and antifungal activity was observed. The bimetallic composites demonstrated greater activity than monometallic composites against these bacterial and fungal species. All bimetallic nanocomposites have shown enhanced, loss due to reflection, loss due to absorption, and the total EMI shielding efficiency at 8 GHz (X-band) and 16 GHz (Ku-band) frequency. All these results ratify, that these newly developed bio nanocomposites are most suitable in many applications, in EMI shielding, nanotechnology, and medical fields.

Recombinant expression and characterization of the endochitinase Chit36-TA from Trichoderma asperellum in Komagataella phaffii for chitin degradation of black soldier fly exuviae.

The natural polymer chitin is an abundant source for valuable N-acetylchitooligosaccharides and N-acetylglucosamine applicable in several industries. The endochitinase Chit36-TA from Trichoderma asperellum was recombinantly expressed in Komagataella phaffii for the enzymatic degradation of chitin from unused insect exuviae into N-acetylchitooligosaccharides. Chit36-TA was purified by Ni-NTA affinity chromatography and subsequently biochemically characterized. After deglycosylation, the endochitinase had a molecular weight of 36 kDa. The optimum pH for Chit36-TA was 4.5. The temperature maximum of Chit36-TA was determined to be 50 °C, while it maintained > 93% activity up to 60 °C. The chitinase was thermostable up to 45 °C and exhibited ~ 50% activity after a 15 min incubation at 57 °C. Chit36-TA had a maximum specific enzyme activity of 50 nkat/mg with a Km value of 289 µM with 4-methylumbelliferyl-N,N',N″-triacetyl-ß-chitotrioside as substrate. Most tested cations, organic solvents and reagents were well-tolerated by the endochitinase, except for SDS (1 mM), Cu2+ (10 mM) and Mn2+ (10 mM), which had stronger inhibitory effects with residual activities of 3, 41 and 28%, respectively. With a degree of hydrolysis of 32% applying colloidal shrimp chitin (1% (w/v)) and 12% on insect larvae (1% (w/v)) after 24 h, the endochitinase was found to be suitable for the conversion of colloidal chitin as well as chitin from black soldier fly larvae into water-soluble N-acetylchitooligosaccharides. To prove scalability, a bioreactor process was developed in which a 55-fold higher enzyme activity of 49 µkat/l and a tenfold higher protein expression of 1258 mg/l were achieved.