Search

Fate of extracellular antibiotic resistance genes (eARGs)—which are widespread in various environments—is dependent on gene persistence. This work examined the previously unconsidered role of envir...

Aim: Controlled release of flufenamic acid by helical V-amylose to achieve enzyme-responsive, targeted release of the cargo drug. Materials & methods: Solid-state cross-polarization magic angle spinning carbon-13 nuclear magnetic resonance (CP/MAS NMR), Fourier transform IR and x-ray diffraction (XRD) analysis validated the entrapment of flufenamic acid inside the helical structure of V-amylose. Scanning electron microscopy (SEM) investigations established the morphology of conjugates in simulated gastric environment (pH 1.2) and simulated intestine media (pH 7.2) containing hydrolyzing enzyme. Results & discussion: V-amylose–flufenamic acid complex displayed a sustained release of flufenamic acid for 12 h with a marked stability in simulated gastric pH, while showing a controlled release of drug in simulated intestine media. Conclusion: The V-amylose–flufenamic acid system achieves intestine-targeted delivery of flufenamic acid. The controlled release of flufenamic acid may ensure minimal ulcerogenicity and application as enteric coatings.

The temporomandibular joint disc is a fibrocartilaginous structure, composed of collagen fibers, elastin fibers, and proteoglycans. Despite the crucial role of elastin fibers in load‐bearing properties of connective tissues, its contribution in temporomandibular joint disc biomechanics has been disregarded. This study attempts to characterize the structural–functional contribution of elastin in the temporomandibular joint disc. Using elastase, we selectively perturbed the elastin fiber network in porcine temporomandibular joint discs and investigated the structural, compositional, and mechanical regional changes through: (a) analysis of collagen and elastin fibers by immunolabeling and transmission electron microscopy; (b) quantitative analysis of collagen tortuosity, cell shape, and disc volume; (c) biochemical quantification of collagen, glycosaminoglycan and elastin content; and (d) cyclic compression test. Following elastase treatment, microscopic examination revealed fragmentation of elastin fibers across the temporomandibular joint disc, with a more pronounced effect in the intermediate regions. Also, biochemical analyses of the intermediate regions showed significant depletion of elastin (50%), and substantial decrease in collagen (20%) and glycosaminoglycan (49%) content, likely due to non‐specific activity of elastase. Degradation of elastin fibers affected the homeostatic configuration of the disc, reflected in its significant volume enlargement accompanied by remarkable reduction of collagen tortuosity and cell elongation. Mechanically, elastase treatment nearly doubled the maximal energy dissipation across the intermediate regions while the instantaneous modulus was not significantly affected. We conclude that elastin fibers contribute to the restoration and maintenance of the disc resting shape and actively interact with collagen fibers to provide mechanical resilience to the temporomandibular joint disc.

Perfluorooctanoate (PFOA) and perfluorooctanesulfonate (PFOS) are two environmentally persistent per- and polyfluoroalkyl substances (PFAS) that have been detected globally in human tissues and fluids. As part of a project investigating the indirect sources of PFOA/PFOS in the environment and engineered systems, this study is concerned with the mechanisms leading to their in vivo generation in terrestrial invertebrates. We demonstrate here the formation of PFOA and PFOS in earthworms (Lumbricus terrestris) from a group of four zwitterionic/cationic polyfluoroalkyl amides and sulfonamides. In bioaccumulation tests, the zwitterionic PFAS compounds were metabolized within 10 days to PFOA/PFOS at yields of 3.4-20.8 mol % by day 21 and several infrequently reported PFAS species for which chemical structures were determined using high-resolution mass spectrometry. Cationic PFAS, on the other hand, were found to be much less metabolizable in terms of the number (n = 2) and yields (0.9-5.1 mol %) of metabolites. Peak-shaped bioaccumulation profiles were frequently observed for the studied PFAS. Residual zwitterionic/cationic PFAS in earthworms were detected at the end of the elimination phase, indicating that not all zwitterionic/cationic PFAS molecules in vivo are available for enzymatic degradation. Finally, the relative importance of different exposure routes (i.e., waterborne and dietary exposure) was investigated.

Summary There are a multitude of resistance strategies that microbes can apply to avoid inhibition by antimicrobials. One of these strategies is the enzymatic modification of the antibiotic, in a process generally termed inactivation. Furthermore, some microorganisms may not be limited to the mere inactivation of the antimicrobial compounds. They can continue by further enzymatic degradation of the compounds' carbon backbone, taking nutritional and energetic advantage of the former antibiotic. This driving force to harness an additional food source in a complex environment adds another level of complexity to the reasonably well‐understood process of antibiotic resistance proliferation on a single cell level: It brings bioprotection into play at the level of microbial community. Despite the possible implications of a resistant community in a host and a lurking antibiotic failure, knowledge of degradation pathways of antibiotics and their connections is scarce. Currently, it is limited to only a few families of antibiotics (e.g. β‐lactams and sulfonamides). In this article, we discuss the fluctuating nature of the relationship between antibiotic resistance and the biodegradation of antibiotics. This distinction mainly depends on the genetic background of the microbe, as general resistance genes can be recruited to function in a biodegradation pathway.

Koji is commonly used in manufacturing Japanese fermented products and promotes enzymatic degradation. In recent years, a seaweed koji has been developed by culturing Aspergillus oryzae on nori Pyropia yezoensis. In the present study, we prepared nori koji by culturing A. oryzae on high- and low-quality noris and then mixing this preparation with additional dried nori. Suitable fermentation conditions for increasing the taste-active components of the nori mixed with nori koji were investigated. The mixture of nori and koji prepared from high-quality nori, with added water, cultured for 120 days at 10 °C, provided the greatest increase (7 times) in free amino acids. The changes in taste after culturing the nori and koji mixture were evaluated using a taste-sensing system. The sourness score of the nori and koji mixture increased significantly, but the scores for other attributes such as bitterness and umami richness did not increase after culture. The present study has demonstrated a clear increase in the free amino acid content of nori and a modification in the taste score by aging the culture with nori koji. These results will encourage the development of ‘aged seaweed,’ a novel value-added product with nutritional and taste elements modified using seaweed koji.

Tuliposides (Pos) are major defensive secondary metabolites in tulip (genus Tulipa), having 4-hydroxy-2-methylenebutanoyl and/or (3S)-3,4-dihydroxy-2-methylenebutanoyl groups at the C-1 and/or C-6 positions of d-glucose. The acyl group at the C-6 position is converted to antimicrobial lactones, tulipalins, by tuliposide-converting enzymes (TCEs). In the course of a survey of tulip tissue extracts to identify novel Pos, we found a minute high-performance liquid chromatography peak that disappeared following the action of a TCE, and whose retention time differed from those of known Pos. Spectroscopic analyses of the purified compound, as well as its enzymatic degradation products, revealed its structure as 5″-O-(6-O-(4′-hydroxy-2′-methylenebutanoyl))-β-d-glucopyranosyl-(2″R)-2″-hydroxymethyl-4″-butyrolactone, which is a novel glucoside ester-type Pos. We gave this compound the trivial name ‘tuliposide G’ (PosG). PosG accumulated in bulbs, at markedly lower levels than 6-PosA (the major Pos in bulbs), but was not found in any other tissues. Quantification of PosG in bulbs of 52 types of tulip, including 30 cultivars (Tulipa gesneriana) and 22 wild Tulipa spp., resulted in the detection of PosG in 28 cultivars, while PosG was present only in three wild species belonging to the subgenus Tulipa, the same subgenus to which tulip cultivars belong, suggesting the potential usefulness of PosG as a chemotaxonomic marker in tulip.

The work is devoted to the research of the enzymatic destruction process of solid waste of the primary winemaking enterprises the introduction of advanced technology of their utilization and creation of optimally balanced feed additive. The processing of grapes produces waste, the storage and accumulation of which in environmental components leads to the loss of valuable resource in the composition of waste and leads to the creation of increased levels of environmental hazards. Enzymatic degradation of cellulose in the composition of waste by the cellulase enzyme through the microorganisms' action is the basis of the biotechnological process. To implement this process, it is necessary to determine the main parameters and modes of the bioconversion process, investigation of which the study is devoted. The basic characteristics of the enzymatic degradation process were determined and data were obtained for the technology improvement of solid waste utilization of the wine industry enterprises. Comparative analysis of bioconversion of different types of waste proves that the cellulose enzymatic degradation is the most effective for the grape pomace in the waste, allows us to obtain valuable feed additive and reduce the environmental danger level.

Disadvantages in the use of polylactic acid (PLA) as a base material for Tissue Engineering applications include the low osteoconductivity of this biomaterial, its acidic degradation and the deficient cellular adhesion on its surface. In order to counteract these drawbacks, calcium carbonate (CaCO3) and β-tricalcium phosphate (Ca3(PO4)2, β-TCP) were proposed in this work as additives of PLA-based support structures. Composite scaffolds (PLA:CaCO3: β-TCP 95:2.5:2.5) manufactured by fused deposition modeling (FDM) were tested under enzymatic degradation using proteinase K enzymes to assess the modification of their properties in comparison with neat PLA scaffolds. The samples were characterized before and after the degradation test by optical microscopy, scanning electron microscopy, compression testing and thermogravimetric and calorimetric analysis. According to the results, the combination of the PLA matrix with the proposed additives increases the degradation rate of the 3D printed scaffolds, which is an advantage for the application of the composite scaffold in the field of Tissue Engineering. The higher degradation rate of the composite scaffolds could be explained by the release of the additive particles and the statistically higher microporosity of these samples compared to the neat PLA ones.

To more precisely control the degradation rate of poly(trimethylene carbonate) (PTMC), self-blending films were prepared via the ternary self-blending of pure PTMC with a molecular weight of 334, 152, and 57 kg/mol. The in vitro enzymolysis degradation of the ternary self-blending films was performed in lipase solutions. The results showed that ternary self-blending could control the degradation of PTMC by adjusting the mass ratio of high/medium/low molecular weight PTMC in the composition, and the PTMC334/PTMC152/PTMC57 films with a mass ratio of 1/4/16 showed mass loss of 85.96% after seven weeks of degradation, while that of PTMC334/PTMC152/PTMC57 films with a mass ratio of 1/1/1 was 96.39%. The former and latter’s degradation rate constant was 13.263 and 23.981%/w, respectively, and the former presented better morphology stability than the latter. The strategy of ternary self-blending could simultaneously bestow PTMC with a lower degradation rate and good morphology stability, indicating that ternary self-blending is an efficient way to control the degradation performance of PTMC more precisely.

Recently, fruit-drying industries are showing great interest in producing dry fruits that preserve a high enzyme content. Therefore, this study aimed to investigate the effect of hot-air convective drying on activity retention of amylase and invertase in dried mango of varieties Sindri, Samar Bahisht (SB) Chaunsa, and Tommy Atkins. Convection drying was conducted under over-flow mode at five temperatures (40, 50, 60, 70, and 80 °C), two air velocities (1.0 and 1.4 m s−1), and constant specific humidity of 10 g kg−1 dry air. The enzymatic degradation data were fitted to the first-order reaction kinetics model, in which the temperature dependence of the rate constant is modelled by the Arrhenius-type relationship. Results showed that the maximum amylase and invertase activity for dried mango of all three varieties was best preserved in samples dried at a temperature of 80 °C and an air velocity of 1.4 m s−1. In contrast, a lower drying temperature and an air velocity of 1.0 m s−1 contributed to a significant decrease (p < 0.05). Exploration of different temperatures and air velocities to save amylase and invertase in dried mango is useful from an industrial point of view, as mango can be a natural dietary source of digestive enzymes to improve digestion.

Background Flonicamid (N-cyanomethyl-4-trifluoromethylnicotinamide, FLO) is a new type of pyridinamide insecticide that regulates insect growth. Because of its wide application in agricultural production and high solubility in water, it poses potential risks to aquatic environments and food chain. Results In the present study, Ensifer adhaerens CGMCC 6315 was shown to efficiently transform FLO into N-(4-trifluoromethylnicotinoyl) glycinamide (TFNG-AM) via a hydration pathway mediated by two nitrile hydratases, PnhA and CnhA. In pure culture, resting cells of E. adhaerens CGMCC 6315 degraded 92% of 0.87 mmol/L FLO within 24 h at 30 °C (half-life 7.4 h). Both free and immobilized (by gel beads, using calcium alginate as a carrier) E. adhaerens CGMCC 6315 cells effectively degraded FLO in surface water. PnhA has, to our knowledge, the highest reported degradation activity toward FLO, Vmax = 88.7 U/mg (Km = 2.96 mmol/L). Addition of copper ions could increase the enzyme activity of CnhA toward FLO by 4.2-fold. Structural homology modeling indicated that residue β-Glu56 may be important for the observed significant difference in enzyme activity between PnhA and CnhA. Conclusions Application of E. adhaerens may be a good strategy for bioremediation of FLO in surface water. This work furthers our understanding of the enzymatic mechanisms of biodegradation of nitrile-containing insecticides and provides effective transformation strategies for microbial remediation of FLO contamination. Supplementary Information The online version contains supplementary material available at 10.1186/s12934-021-01620-4., Highlights E. adhaerens efficiently degrades the insecticide FLO via a hydration pathway.Both free and immobilized cells effectively degrade FLO in surface water.E. adhaerens nitrile hydratases CnhA and PnhA both hydrolyze FLO to TFNG-AM.PnhA has, to our knowledge, the highest reported degradation activity toward FLO.The key residue (β-Glu56) may cause a significant difference in two NHase activities. Supplementary Information The online version contains supplementary material available at 10.1186/s12934-021-01620-4.

During development, the fruit of some paprika (Capsicum annuum L.) cultivars shows a change in color from green to dark purple (e.g., ‘Mavras’) or lilac (e.g., ‘Tequila’). However, this purple coloration is rare among paprika cultivars and disappears in ripened fruit, which are red. Therefore, we investigated the mechanism causing this color change in the cultivars Mavras and Tequila to better understand how purple ripened fruit could be generated. High-performance liquid chromatography (HPLC) analyses of the anthocyanin contents of the fruit indicated that anthocyanin was undetectable in green fruit, accumulated in dark purple or lilac ones, and then decreased again in red ones in both cultivars. Furthermore, expressions of most of the analyzed anthocyanin biosynthesis–related genes and genes for their transcription factors increased in dark purple or lilac fruit and decreased in red ones, i.e., it was synchronized with the changes in anthocyanin contents. Furthermore, anthocyanin degradation activity as a result of peroxidases was detected at all stages but increased when the lilac or dark purple color started to fade. Thus, the development of purple coloration is caused by increased anthocyanin biosynthesis, whereas the fading of this coloration is a result of both a decrease in anthocyanin biosynthesis and an increase in anthocyanin degradation. At the ripening stage, the green pigment (chlorophyll) contents decreased, whereas the red pigment (carotenoid, particularly capsanthin) contents increased. However, these timings did not completely coincide with the timing of anthocyanin degradation, suggesting that the content of each pigment is individually regulated, and so purple, green, and red coloration could be freely expressed in mature paprika fruit.

O-antigens of Gram-negative bacteria modulate the interactions of bacterial cells with diverse external factors, including the components of the immune system and bacteriophages. Some phages need to acquire specific adhesins to overcome the O-antigen layer. For other phages, O-antigen is required for phage infection. In this case, interaction of phage receptor binding proteins coupled with enzymatic degradation or modification of the O-antigen is followed by phage infection. Identification of the strategies used by newly isolated phages may be of importance in their consideration for various applications. Here we describe an approach based on screening for host LPS alterations caused by selection by bacteriophages. We describe an optimized LPS profiling procedure that is simple, rapid and suitable for mass screening of mutants. We demonstrate that the phage infection strategies identified using a set of engineered E. coli 4 s mutants with impaired or altered LPS synthesis are in good agreement with the results of simpler tests based on LPS profiling of phage-resistant spontaneous mutants.

Human milk oligosaccharides (HMOs) have attracted much attention in recent years not only as a prebiotic factor, but in particular as an essential component in infant nutrition related to their impact in innate immunity. The backbone structures of complex HMOs generally contain single or repetitive lacto-N-biose (type 1) or lactosamine (type 2) units in either linear or branched chains extending from a lactose core. While all known branched structures originate from 3,6-substitution of the lactosyl core galactose, we here describe a new class of HMOs that tentatively branch at terminal galactose of 6’-galactosyllactose. Another novel feature of this class of HMOs was found in linear oligo-galactosyl chains linked to one of the N-acetylglucosamine (GlcNAc) branches. The novel structures exhibit general formulas with hexose vs. hexosamine contents of 5/2 to 8/2 and can be designated as high-galactose (HG)-HMOs. In addition, up to three fucosyl residues are linked to the octa- to dodecasaccharides, which were detected in two human milk samples from Lewis blood group defined donors. Structural analyses of methylated glycans and their alditols comprised MALDI mass spectrometry, ESI-(CID)MS and linkage analyses by GC-MS of the derived partially methylated alditol acetates. Enzymatic degradation by application of β1-3,4-specific galactosidase supported the presence of terminal galactose linked β1−6 to one of the two GlcNAc branches.

Gracilariopsis lemaneiformis polysaccharides (GLP) were degraded using pectinase, glucoamylase, cellulase, xylanase, and β-dextranase into low-molecular-weight polysaccharides, namely, GPP, GGP, GCP, GXP, and GDP, respectively, and their antioxidant capacities were investigated. The degraded GLP showed higher antioxidant activities than natural GLP, and GDP exhibited the highest antioxidant activity. After the optimization of degradation conditions through single-factor and orthogonal optimization experiments, four polysaccharide fractions (GDP1, GDP2, GDP3, and GDP4) with high antioxidant abilities (hydroxyl radical scavenging activity, DPPH radical scavenging activity, reduction capacity, and total antioxidant capacity) were obtained. Their cytoprotective activities against H2O2-induced oxidative damage in human fetal lung fibroblast 1 (HFL1) cells were examined. Results suggested that GDP pretreatment can significantly improve cell viability, reduce reactive oxygen species and malonaldehyde levels, improve antioxidant enzyme activity and mitochondria membrane potential, and alleviate oxidative damage in HFL1 cells. Thus, the enzyme degradation of GLP with β-dextranase can significantly improve its antioxidant activity, and GDP might be a suitable source of natural antioxidants.

In this work, the versatility of layer-by-layer technology was combined with the magnetic response of iron oxide nanobeads to prepare magnetic mesostructures with a degradable multilayer shell into which a dye quenched ovalbumin conjugate (DQ-OVA) was loaded. The system was specificallydesigned to prove the protease sensitivity of the hybrid mesoscale system and the easy detection of the ovalbumin released. The uptake of the nanostructures in the breast cancer cells was followed by the effective release of DQ-OVA upon activation via the intracellular proteases degradation of the polymer shells. Monitoring the fluorescence rising due to DQOVA digestion and the cellular dye distribution, together with the electron microscopy studying, enabled us to track the shell degradation and the endosomal uptake pathway that resulted in the release of the digested fragments of DQ ovalbumin in the cytosol.

Synthesis, physicochemical characterization, and the enzymatic degradation of the amphiphilic miktoarm star-shaped polymers is reported herein. First, star-shaped macroinitiators, based on N,N′-dimethylaminoethyl methacrylate (DMAEMA) and glycerol dimethacrylate (GDMA) ((PDMAEMA)n-PGDMA), were synthesized. Due to the presence of hydroxyl groups in the macroinitiator core, polyesters such as poly(ɛ-caprolactone) (P(ɛ-CL)), polylactide (PLA) and poly(lactide-co-glycolide) (PLGA) were synthesized using ring opening polymerization (ROP). Comprehensive degradation studies on enzymatic degradation, using a lipase from Pseudomonas cepacia, were performed. Enzymatic degradation was monitored by weight measurements and nuclear magnetic resonance spectroscopy (1H NMR). The fastest degradation rate was observed for the polymer with the lowest molecular weight. Amphiphilic miktopolymers may find application as biomaterials for long- or mid-term period drug-delivery systems.

Background: Sugarcane is an important agro-industrial crop as a worldwide manufacturing plant. Scientifically it is the so-called Saccharum officinarum. Sugarcane juice is dark brown golden in color with medium flavor intensity contains caramel, butterscotch and green flavor and sucrose hydrolyzed to an equimolar mixture of glucose and fructose providing instant energy transported by fluid thermodynamics. The expected research based on a new implementation of harmful pathogens against roadside sellers of sugarcane juices. The work is currently reviewed and renewed for automated analysis techniques with verified street juice samples at Guwahati city in Assam. Following spring to summer seasons, the favorite juice to people of all ages in India is supposed to health-threatening for public health concerns. Respective doctors and Practitioner carry out their treatment against this pathogen along with antibiotic supplements unknowingly the etiological source about UTI or GI tract infections. Methods: Here researchers finding (Chowdhury et al., 2019) are to analyze and categorize the biochemical fermentation activities of pathogens with significant enzyme degrading analyzed from sugarcane juice against gut acting probiotics, responsible for food toxicity and intestinal disorder. Diagnostic microbial activities in well diffusion studies are followed by automated analysis for significant provoking of health status.Result: Epidemiological studies were concerned to identify risk factors through the comparison of incidence and prevalence between groups exposed and not exposed to risk factors. Probabilities of disease occurrence can be compared using measures of strength of association or potential impact. The studies are compared with the graph and its data implementations to find out the significant level for beneficial sources.

[EN] Alginate is a polysaccharide used extensively in biomedical applications due to its biocompatibility and suitability for hydrogel fabrication using mild reaction chemistries. Though alginate has commonly been crosslinked using divalent cations, covalent crosslinking chemistries have also been developed. Hydrogels with tuneable mechanical properties are required for many biomedical applications to mimic the stiffness of different tissues. Here, we present a strategy to engineer alginate hydrogels with tuneable mechanical properties by covalent crosslinking of a norbornene-modified alginate using ultraviolet (UV)-initiated thiol-ene chemistry. We also demonstrate that the system can be functionalised with cues such as full-length fibronectin and protease-degradable sequences. Finally, we take advantage of alginate's ability to be crosslinked covalently and ionically to design dual crosslinked constructs enabling dynamic control of mechanical properties, with gels that undergo cycles of stiffening-softening by adding and quenching calcium cations. Overall, we present a versatile hydrogel with tuneable and dynamic mechanical properties, and incorporate cell-interactive features such as cell-mediated protease-induced degradability and full-length proteins, which may find applications in a variety of biomedical contexts., This researchwas funded by Deutsche Forschungsgemeinschaft (DFG) grant CI 203/1-2, DFG Emmy Noether grant (CI 203/2-1) and the Berlin-Brandenburg School for Regenerative Therapies (BSRT) Extension grant. This work was also supported by an EPSRC Program Grant (EP/P001114/1) and from the UK Regenerative Medicine Platform "Acellular/Smart Materials-3D Architecture" (MR/R015651/1). This research was also supported by the Spanish State Research Agency (AEI) through the PID2019-106000RB-C21/AEI/10.13039/501100011033 project (including the FEDER funds). The article processing charge was funded by the Max Planck Society.

Hydrolytic degradation of poly(4-hydroxybutyrate) (P4HB) films has been studied considering media of different pH values (i.e., 3, 7 and 10) and temperatures (i.e., 37 and 55 °C). Enzymatic degradation has also been evaluated at physiological conditions using two different lipases: Pseudomonas cepacia and Rhizopus oryzae. Different bulk and surface erosion mechanisms with random chain scissions and successive removal of monomer units have been supported through weight loss measurements, molecular weight determinations by GPC and NMR spectroscopy and changes on thermal properties by DSC. Thermal annealing during exposure to different media and even degradation influenced on the melting temperature and crystallinity of samples, as well as on the lamellar geometrical parameters as evaluated by SAXS. Enzymatic degradation was ideal to selectively eliminate the amorphous regions and highlight the spherulitic morphology. Presence of ringed textures were therefore evident in bright field optical micrographs in addition to SEM images, namely observations under polarized light was not necessary to distinguish the presence of banded spherulites. Rhizopus oryzae was revealed to be the most suitable enzyme to crop out the P4HB spherulites that form part of the initial smooth surfaces of solvent casting films. After determining the appropriate activity and exposure time, the presence of rings constituted by cooperative C-shaped edge-on lamellae and flat-on lamellae was highlighted.

White-rot fungus Rigidoporus sp. FMD21 is a lignin-modifying enzyme producing fungus that can degrade dioxin. Extracellular enzymes from FMD21 include laccase and manganese peroxidase which are promising enzymes for myco-remediation because of their wide substrate specificity and mild catalysis conditions. The FMD21 genome was sequenced using Ion Torrent technology and consists of 38.98 Mbps with a GC content of 47.4 %. Gene prediction using Augustus with Basidiomycota reference setting resulted in 8245 genes. Functional gene annotations were carried out by using several programs and databases. We focused on laccase and ligninolytic peroxidase genes, which are most likely involved in the degradation of aromatic pollutants. The genome of FMD21 contains 12 predicted laccase genes (10 out of 12 predicted as full length) and 13 putative ligninolytic peroxidases which were annotated as MnP or versatile peroxidases. Four predicted laccases showed a higher than 65 % binding chance to 2,3,7,8-TCDD with the highest at 72 % in in silico docking analysis. Heterologous expressed laccases showed activity towards three tested substrates included ABTS, guaiacol and 2,6-DMP. ABTS displayed two-stage oxidation which differed from natural FMD21 laccases. 2,3,7,8-TCDD was degraded by 50 % after two weeks of enzymatic treatment by three out of five laccase isozymes which were natural laccases secreted by FMD21. In this study, we provide direct evidence for the 2,3,7,8-TCDD biodegradation capability of fungal laccases.

Ethanol is one of the bioenergy sources with low environmental and high efficiency impact. The aim of this study was to screen for the bacterial isolate capable of degrading starch, investigate the enzymatic hydrolysis and fermentation of corn bran through submerged fermentation using co-culture technique for bioethanol production. The isolate was identified using 16S rRNA sequence technique as Pseudomonas aeruginosa AU4738. Corn bran was used as substrates with and without garlic powder (Allium sativum L.) as activator and subsequently optimized for production of bioethanol. Reducing sugar from the hydrolysate and ethanol concentration of the distillate were analyzed using spectrophotometry and gas chromatography mass spectrometry techniques respectively. There was an increase in glucose concentration (23.8% and 17.8%) in the culture medium with and without activator at 48 h respectively but steadily decreased from 72 h to 168 h. Maximum ethanol concentration obtained in substrate culture with activator was 35% higher compared with that without activator at 120 h fermentation time. Thus a cheap, renewable and readily available agricultural waste has been effectively utilized as substrate for bioethanol production and incorporation of activator also had significant effect on the viability of fermenting organisms thus subjugating the intolerance of alcohol concentration.

Most proteins in cell and tissue lysates are soluble. We show here that in lysate from human neurons, more than 1,300 proteins are maintained in a soluble and functional state by association with endogenous RNA, as degradation of RNA invariably leads to protein aggregation. The majority of these proteins lack conventional RNA‐binding domains. Using synthetic oligonucleotides, we identify the importance of nucleic acid structure, with single‐stranded pyrimidine‐rich bulges or loops surrounded by double‐stranded regions being particularly efficient in the maintenance of protein solubility. These experiments also identify an apparent one‐to‐one protein‐nucleic acid stoichiometry. Furthermore, we show that protein aggregates isolated from brain tissue from Amyotrophic Lateral Sclerosis patients can be rendered soluble after refolding by both RNA and synthetic oligonucleotides. Together, these findings open new avenues for understanding the mechanism behind protein aggregation and shed light on how certain proteins remain soluble., This study shows that RNA plays an important and direct role in maintaining protein solubility and preventing protein aggregation.

International audience; In the present study, absorption in an organic solvent followed by enzymatic biodegradation by cross-linked peroxidase from Brassica rapa (BRP-CLEAs) was successfully applied as a solution for toluene removal from waste-air in batch experiments. Silicone oil (PDMS 47V20) showed a high capacity for toluene absorption, regarding its air-PDMS partition coefficient (2.08 Pa m(3) mol(-1)) when compared to air-water (666 Pa m(3) mol(-1)). Box-Behnken restricted duplicate design was employed for the optimization of the enzymatic degradation of toluene in PDMS. Effects of four independent variables, namely, BRP-CLEAs activity, toluene and hydrogen peroxide concentrations and sonication, were studied by choosing two responses conversion yield and initial rate of reaction. Analysis of the results showed that both hydrogen peroxide and toluene concentrations presented the largest effect on the process while sonication had no effect. On the other hand, the coefficient of determination R-2 and the adjusted R-2 were 0.828 and 0.770 for conversion and 0.919 and 0.911 for initial rate, respectively. The optimum conversion determined was 60% of toluene removal.

As a biomaterial, silk presents unique features with a combination of excellent mechanical properties, biocompatibility and biodegradability. The biodegradability aspects of silk biomaterials, especially with options to control the rate from short (days) to long (years) time frames in vivo, make this protein-based biopolymer a good candidate for developing biodegradable devices used for tissue repairs and tissue engineering, as well as medical device implants. Silk materials, including native silk fibers and a broad spectrum of regenerated silk materials have been investigated in vitro and in vivo to demonstrate degradation by proteolytic enzymes. In this mini review, we summarize the findings on these studies on the enzymatic degradation of Bombyx mori (B. mori) silk materials. We also present a discussion on the factors that dictate the degradation properties of silk materials. Finally, in future perspectives, we highlight some key challenges and potential directions toward the future study of the degradation of silk materials.

Basidiomycota species that cause “brown rot” or “white rot” of wood are the primary organisms involved in depolymerization of the two most abundant polymeric materials on earth: cellulose and lignin. Ascomycota species are also involved in the deconstruction of wood, and although less ubiquitous compared to the Basidiomycota decays, these Ascomycota species cause a “soft rot” decay of wood that is important in some environments. Wood decay fungi are important from the standpoint of cycling and sequestration of recalcitrant lignocellulose carbon, and from an economic standpoint, they also cause significant destruction of the built environment. This chapter will review these decay fungi but also will overview Ascomycota molds and staining (stain) fungi that live on the surface or attack parenchyma of harvested wood and that typically do not cause decay or structural deterioration of timber. Over the last 70 years, several genera and species of fungi discussed in this chapter, and the extracellular enzymes and metabolic systems from these fungi, have been examined for potential applications in industrial processing, particularly in biorefinery applications.

Ascomycete fungi have a long history of use in biotechnology, in particular with respect to the production of enzymes and metabolites. A well-developed area is the production of enzymes to degrade or modify plant-based polymers, in particular polysaccharides. However, recent insights due to the availability of genome sequences have revealed a much larger potential in these fungi that was previously assumed. Several studies have made use of this information, but many more opportunities await to be exploited. In contrast, enzymatic degradation or modification of algal polysaccharides has received much less attention and is only recently becoming a booming research field. In this chapter, we will review the current knowledge on enzymes from ascomycete fungi involved in degradation and modification of plant and algal polysaccharides. In addition, we will discuss the potential of omics-based studies to further advance this field and increase the efficiency of obtaining valuable products from algal and plant biomass.

Alginate consists of long chain polysaccharides, which has high molecular weight and viscosity. This high viscosity limits its use as a matrix in spray drying encapsulation. Hence, it needs viscosity modification. The viscosity of polysaccharides can be decreased by breaking the large molecule into smaller monomer. Alginate is a copolymer of α (1 → 4)-linked l-guluronic acid and β (1→4)-linked d-mannuronic acid. This β (1→4) linkage is a target in degradation using cellulase. This work aimed to study viscosity of alginate enzymatic degradation by using cellulase under variation of alginate and cellulase concentrations. Direct reducing sugar (DRS) was observed a long with the degradation. The viscosity dropped from 5.56 ml/g to 1.59 ml/g in degradation using 350 ppm cellulase of 3% alginate for 300 min. This degradation resulted in increasing of DRS from 0.105 mg/ml to 0.441 mg/ml. This result highlights the potency of enzymatic degradation to be applied in preparing viscosity of alginate to be applied as matrix of spray drying encapsulation method.

This work examined the effects of filler content and chemical treatment on the biodegradation of poly(lactic acid) (PLA)/coconut shell (CS) biocomposites in a diastase enzyme-containing buffer medium. CS was treated with two distinct chemical treatments: maleic acid and silanation with 3-aminopropyltriethoxysilane (3-APE). The CS was incorporated into PLA composites and their biodegradation patterns were studied. Both of the treated PLA/CS biocomposites exhibited lower biodegradation rates than the untreated biocomposites due to their enhanced interfacial adhesion, which reduced the area exposed to enzyme hydrolysis. Scanning electron micrographs taken after 30 days of biodegradation displayed surface roughening on both of the treated biocomposites, with fewer voids compared to the untreated biocomposites. The differential scanning calorimetry indicated that the glass transition temperature and melting temperature values of the treated biocomposites increased but that crystallinity declined. The crystallization temperature peak apparently disappeared due to the polymer chain alignment and rearrangement of the shorter PLA chains caused by the degradation. Fourier transform infrared analysis revealed the structural changes in the biocomposites after biodegradation, indicating the presence of soluble lactic acid as was confirmed by ultraviolet–visible spectroscopy analysis.

Cow's milk is one of the most common allergenic foods. Cow's milk allergy is mainly an IgE-mediated hypersensitivity reaction, and the major allergens from cow's milk have been found to be caseins, β-lactoglobulin, and α-lactalbumin. Several peptides derived from bovine casein are known allergens in cow's milk. To reduce their allergenicity, these proteins can be degraded by food-grade peptidases. We succeeded in detection of two peptides, VLPVPQK and FFVAPFPEVFGK, from bovine casein-derived allergen peptides by using an ion trap LC-MS apparatus. This study focuses on the synergistic effects of Streptomyces aminopeptidases belonging to the M1, M24, and M28 families on the degradation of the allergen peptides. From these results, we demonstrated that the combination of M1 and M24 aminopeptidases was the most effective for degrading the abovementioned allergenic peptides.

The development and introduction of new dietary protein sources has the potential to improve food supply sustainability. Understanding the potential allergenicity of these new or modified proteins is crucial to ensure protection of public health. Exposure to new proteins may result in de novo sensitization, with or without clinical allergy, or clinical reactions through cross-reactivity. In this paper we review the potential of current methodologies (in silico, in vitro degradation, in vitro IgE binding, animal models and clinical studies) to address these outcomes for risk assessment purposes for new proteins, and especially to identify and characterise the risk of sensitization for IgE mediated allergy from oral exposure. Existing tools and tests are capable of assessing potential crossreactivity. However, there are few possibilities to assess the hazard due to de novo sensitization. The only methods available are in vivo models, but many limitations exist to use them for assessing risk. We conclude that there is a need to understand which criteria adequately define allergenicity for risk assessment purposes, and from these criteria develop a more suitable battery of tests to distinguish between proteins of high and low allergenicity, which can then be applied to assess new proteins with unknown risks. © 2017 The Authors Chemicals/CAS: immunoglobulin E, 37341-29-0

Textile and food industries wastewaters are characterized by a strong coloration due to use of dyes, among which azo dyes are the most frequently used. These compounds are harmful and their discharge without an adequate treatment causes a negative impact on the environment. Given that conventional biological treatment processes are usually not sufficient to degrade azo dyes, other treatment alternatives must be studied. Enzymes have demonstrated their effectiveness to degrade several types of recalcitrant compounds. In this study, enzymatic degradation of Crystal Ponceau 6R (CP6R) azo dye by turnip (Brassica rapa) peroxidase was assessed in the presence of 100 μM redox mediator (1-hydroxybenzotriazole). The treatment performance was evaluated at different pH values (3, 4, 5, 6 and 7) and the most appropriate pH and contact time were determined. Moreover, a kinetic model of the enzyme-catalyzed reaction was developed by mathematical description of the degradation mechanism, being further validated with experimental data. For the estimation of the kinetic parameters and model validation, a central composite design of type “start points” was made. The results showed that the enzymatic treatment was very effective. CP6R dye degradation higher than 97% was achieved in less than 1 min of reaction at the optimum pH (i.e., 4), at which the maximum reaction rate (43 μM min−1) was obtained. At the other tested pH values, degradation performance was substantially lower (

The field of enzymatic degradation of lignocellulose is actively growing and the recent updates of the last few years indicate that there is still much to learn. The growing number of protein sequences with unknown function in microbial genomes indicates that there is still much to learn on the mechanisms of lignocellulose degradation. In this review, a summary of the progress in the field is presented, including recent discoveries on the nature of the structural polysaccharides, new technologies for the discovery and functional annotation of gene sequences including omics technologies, and the novel lignocellulose-acting enzymes described. Novel enzymatic activities and enzyme families as well as accessory enzymes and their synergistic relationships regarding biomass breakdown are described. Moreover, it is shown that all the valuable knowledge of the enzymatic decomposition of plant biomass polymers can be employed towards the decomposition and upgrading of synthetic polymers, such as plastics.

The enzymatic degradation of blood clots, fibrinolysis, is an important part of a healthy hemostatic system. If intrinsic fibrinolysis is ineffective, thrombolysis—the clinically-induced enzymatic degradation of blood clots—may be necessary to treat life-threatening conditions. In this review, we discuss recent models of fibrinolysis and thrombolysis, and open questions that could be resolved through modeling and modeling-experimental collaboration. In particular, we focus on 2- and 3-dimensional models that can be used to study effects of fibrin network structure and realistic blood vessel geometries on the phenomena underlying lytic outcomes. Significant open questions such as the role of clot contraction, network and inherent fiber tension, and fibrinolytic inhibitors in lysis could benefit from mathematical models aimed at understanding the underlying biological mechanisms.

The success of the regenerative process resulting from the implantation of a scaffold or a tissue-engineered structure into damaged tissues depends on a series of factors, including, crucially, the biodegradability of the implanted materials. The selection of a scaffold with appropriate biodegradation characteristics allows for synchronization of the degradation of the construct with the processes involved in new tissue formation. Thus, it is extremely important to characterize the biodegradation properties of potential scaffold materials at the stage of in vitro studies. We have analyzed the biodegradation of hybrid fibrin–collagen scaffolds in both PBS solution and in trypsin solution and this has enabled us to describe the processes of both their passive and enzymatic degradation. It was found that the specific origin of the collagen used to form part of the hybrid scaffolds could have a significant effect on the nature of the biodegradation process. It was also established, during comparative studies of acellular scaffolds and scaffolds containing stem cells, that the cells, too, make a significant contribution to changes in the biodegradation and structural properties of such scaffolds. The study results also provided evidence indicating the dependency between the pre-cultivation period for the cellular scaffolds and the speed and extent of their subsequent biodegradation. Our discussion of results includes an attempt to explain the mechanisms of the changes found. We hope that the said results will make a significant contribution to the understanding of the processes affecting the differences in the biodegradation properties of hybrid, biopolymer, and hydrogel scaffolds.

Blend films of Poly (trimethylene carbonate) (PTMC) homopolymers with different molecular weights were prepared. The in vitro enzymatic degradation tests were carried out in lipase solutions to investigate the effect of self-blending on the degradation performance of PTMC. It was observed that the mass loss decreased as the proportion of low molecular weight PTMC in the blend film increased. And the sample's mass loss with the mass ratio of PTMC334/PTMC57=1/16 reached almost 30% after seven weeks, which corresponded to the minimum degradation rate constant k value of 5.219. It is also indicated by the changes in weight as well as the macroscopic and microscopic morphology. Furthermore, during the degradation cycle, the thickness of the film also became thinner with the mass loss, confirming the surface erosion degradation mechanism of the PTMC blend films. The conclusion that PTMC does not produce acidic degradation products has also been confirmed. Finally, decreases in glass transition temperature (Tg) and thermal stability were found along with the film degraded, attributing to the accumulation of low molecular weight degradation products on the surface. In brief, the long-acting application of PTMC in biomedical fields could be satisfied by precisely tailoring the degradation properties via its self-blending with different molecular weights.

This study deals with the design and comprehensive evaluation of novel hydrogels based on whey protein isolate (WPI) for tissue regeneration. So far, WPI has been considered mainly as a food industry by-product and there are very few reports on the application of WPI in tissue engineering (TE). In this work, WPI-based hydrogels were modified with bioactive glass (BG), which is commonly used as a bone substitute material. Ready-to-use, sterile hydrogels were produced by a simple technique, namely heat-induced gelation. Two different concentrations (10 and 20% w/w) of sol–gel-derived BG particles of two different sizes (2.5 and

A material whose enzymatic degradation is initiated by an external stimulus is of great importance from the viewpoint of practical biodegradable polymers. In this context, we developed a method that controls the initiation of enzymatic degradation by light. 4,4′-Didecyloxy-3-methylazobenzene (Azo), which melts upon exposure to UV radiation, was coated onto biodegradable poly(L-lactide) and the enzymatic degradation of the polymer by proteinase K was successfully initiated and controlled by tuning the solid–liquid transition of the azo-based compound. This stimulus-triggered enzymatic degradation is a significant contribution to the development of lifetime-controlled biodegradable materials.

Enzymes are promising candidates for removing environmental carbon nanotubes (CNTs). However, pristine CNTs are difficult to degrade by plant or animal enzymes, such as horseradish peroxidase (HRP) and lactoperoxidase (LPO), and functionalization is often needed for CNT biodegradation. By constructing 12 systems comprising two well-known CNT-degrading enzymes (HRP and LPO) with or without carboxylated or pristine single-walled CNTs (SWCNTs), we answer questions as to how functionalization changes the energetic properties and enzymatic conformations to facilitate the occurrence of SWCNT degradation and how these analysed enzymes respond to different carboxylated SWCNTs using molecular dynamics (MD) simulations. Significant conformational changes in the enzymes were found after the SWCNTs were carboxylated, and functionalization improved the structural stability of enzyme–substrate complexes. Numerous carboxylated SWCNTs induced a similar variation trend of global conformations for both HRP and LPO at the late stages, but their cavity volumes showed totally different change patterns with time. Our study provides a molecular-level understanding of functionalization's roles in SWCNT biodegradation and a molecular basis to develop more biocompatible and biodegradable CNTs.

Aging is an unavoidable progression in everyone's life and influenced by lifestyle, genetic as well as environmental factors. Herbal and plant extracts are used as antiaging since ancient times, but the evidence are still limited. Recent developments in antiaging investigation anticipated the use of natural products as the main ingredient in the formulations. Hence, this presentation focused to highlight the importance of twelve most popular medicinal plant extracts that have reported to have skin aging prevention potential. All these natural product extracts have a capacity to scavenge free radicals and defend skin matrix over the inhibition of enzymatic degradation. Some of the extracts promotes collagen synthesis in the skin and also affect the skin tightness and elasticity. However, the use of natural product extracts as an antiaging and anti-wrinkling it should be further explored using a wide range of in-vitro and in-vivo models to confirm its safety and efficacy before proceeding into the development of cosmetic products.

This review focuses on fibrin, starting from biological mechanisms (its production from fibrinogen and its enzymatic degradation), through its use as a medical device and as a biomaterial, and finally discussing the techniques used to add biological functions and/or improve its mechanical performance through its molecular engineering. Fibrin is a material of biological (human, and even patient's own) origin, injectable, adhesive, and remodellable by cells; further, it is nature's most common choice for an in situ forming, provisional matrix. Its widespread use in the clinic and in research is therefore completely unsurprising. There are, however, areas where its biomedical performance can be improved, namely achieving a better control over mechanical properties (and possibly higher modulus), slowing down degradation or incorporating cell‐instructive functions (e.g., controlled delivery of growth factors). The authors here specifically review the efforts made in the last 20 years to achieve these aims via biomimetic reactions or self‐assembly, as much via formation of hybrid materials.

In situ tissue engineering is a technology in which non-cellular biomaterial scaffolds are implanted in order to induce local regeneration of replaced or damaged tissues. Degradable synthetic electrospun scaffolds are a versatile and promising class of biomaterials for various in situ tissue engineering applications, such as cardiovascular replacements. Functional in situ tissue regeneration depends on the balance between endogenous neo-tissue formation and scaffold degradation. Both these processes are driven by macrophages. Upon invasion into a scaffold, macrophages secrete reactive oxygen species (ROS) and hydrolytic enzymes, contributing to oxidative and enzymatic biomaterial degradation, respectively. This study aims to elucidate the effect of scaffold microarchitecture, i.e., μm-range fiber diameter and fiber alignment, on early macrophage-driven scaffold degradation. Electrospun poly-ε-caprolactone-bisurea (PCL-BU) scaffolds with either 2 or 6 μm (Ø) isotropic or anisotropic fibers were seeded with THP-1 derived human macrophages and cultured in vitro for 4 or 8 days. Our results revealed that macroph age-induced oxidative degradation in particular was dependent on scaffold microarchitecture, with the highest level of ROS-induced lipid peroxidation, NADPH oxidase gene expression and degradation in the 6 μm Ø anisotropic group. Whereas, biochemically polarized macrophages demonstrated a phenotype-specific degradative potential, the observed differences in macrophage degradative potential instigated by the scaffold microarchitecture could not be attributed to either distinct M1 or M2 polarization. This suggests that the scaffold microarchitecture uniquely affects macrophage-driven degradation. These findings emphasize the importance of considering the scaffold microarchitecture in the design of scaffolds for in situ tissue engineering applications and the tailoring of degradation kinetics thereof.

Indoleamine 2,3-dioxygenase (IDO) is an immunomodulating enzyme that is overexpressed in many cancers with poor prognosis. IDO suppresses T cell immunity by catabolizing tryptophan into kynurenine (KYN), which induces apoptosis in T effector cells and enhances T regulatory cells, providing a powerful immunosuppressive mechanism in tumors. Thus, major efforts for developing IDO inhibitors have been undertaken. Among them, 1-Methyl-l-Tryptophan (MLT) and 1-Methyl-d-Tryptophan (MDT) effectively inhibit IDO in preclinical tumor models and the latter is under clinical evaluation. However, both MLT and MDT present poor pharmacokinetics, with the maximum serum concentration being below their 50% inhibitory concentration value. Herein, we have developed polymeric IDO inhibitors based on MLT, which can release active MLT after enzymatic degradation, toward establishing superior antitumor immunotherapies. These polymers were prepared by ring opening polymerization of an N-phenyl carbamate (NPC) derivative of MLT that was synthesized by carbamylation with diphenyl carbonate. By using &omega, amino-poly(ethylene glycol) (PEG-NH2) as the macroinitiator, we prepared amphiphilic PEG-poly(MLT) block copolymers, which self-assembled into polymeric micelles in aqueous conditions. The PEG-poly(MLT) block copolymers could be readily degraded by chymotrypsin and the micelles were able to reduce the levels of KYN in activated macrophages. These results provide a strong rationale for pursuing MLT-based polymeric micelles as tumor-targeted prodrug systems.

Enzymatic degradation of locust bean gum provides a Manβ(1 → 4)Man disaccharide, which may be converted into the core Manβ(1 → 4)GlcNAc disaccharide unit of all N-glycans via conversion to a 2-iodo-glycosyl azide, and Lafont rearrangement. The Manβ(1 → 4)GlcNAc disaccharide may be used as a key intermediate for elaboration into more complex N-glycan structures providing a route to N-glycan oxazolines as donor substrates for ENGase enzymes that is considerably shorter than those reported previously.

The aim of this article was to clarify which type of dye chromophores could be decolorized efficiently with the use of laccase enzyme. For this purpose, enzymatic degradation of different type of dye classes (4 reactive, 2 acid and 1 basic dye) having various chromophore groups was investigated by using commercial laccase from Cerrena unicolor. It was observed that the chromophore structure of dye is very important on enzymatic color removal efficiency. According to the experimental results, it was found that color removal efficiencies (20 mg/L initial dye) were 98.7% for RB220 (0.1 g/L enzyme after 6 h), 95.1% for RB19 (0.1 g/L enzyme after 48 h), 90.8% for AR42 (0.1 g/L enzyme after 48 h) while they were 60.9% for AR114 (0.25 g/L enzyme), 58.6% for RB21 (0.5 g/L enzyme), 39.7% for RR239 (0.25 g/L enzyme) even after seven days. As a result, it can be said that the highest decolorization rate was achieved for the reactive dye having formazan copper complex (RB220) chromophore. On the other hand, the enzymatic degradation of basic dye (BB9) was found to be rather difficult compared to the acid and reactive dyes used in this study and the maximum color removal was 42.8% after seven days.