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Abstract: Paper and pulp industry effluent was enzymatically hydrolysed using crude cellulase enzyme (0.8–2.2FPU/ml) obtained from Trichoderma reesei and from the hydrolysate biohydrogen was produced using Enterobacter aerogenes. The influence of temperature and incubation time on enzyme production was studied. The optimum temperature for the growth of T. reesei was found to be around 29°C. The enzyme activity of 2.5FPU/ml was found to produce about 22g/l of total sugars consisting mainly of glucose, xylose and arabinose. Relevant kinetic parameters with respect to sugars production were estimated using two fraction model. The enzymatic hydrolysate was used for the biohydrogen production using E. aerogenes. The growth data obtained for E. aerogenes were fitted well with Monod and Logistic equations. The maximum hydrogen yield of 2.03mol H2/mol sugar and specific hydrogen production rate of 225mmol of H2/g cell/h were obtained with an initial concentration of 22g/l of total sugars. The colour and COD of effluent was also decreased significantly during the production of hydrogen. The results showed that the paper and pulp industry effluent can be used as a substrate for biohydrogen production. [Copyright &y& Elsevier]

There have been limited studies on the potential use of enzymes for enhancing the dewaterability of biosludge. The mechanisms for such enhancement have not been investigated despite the environmental advantages of using enzymes over synthetic polymers for biosludge conditioning. In order to find enzymes with this potential, a screening of commercially available enzymes was carried out using capillary suction time to assess biosludge dewaterability. The only enzyme that showed dewatering improvements in the screening tests was a lysozyme which reduced the capillary suction time by 36% and increased the cake solids content from 5.6 to 8.9 DS%. Lysozyme aided in the flocculation of particles reducing the polymer demand from 11% to 6%. Active and inactive lysozyme exhibited a similar ability for enhancing sludge dewatering, indicating that the conditioning mechanism of lysozyme is similar to that of a flocculant. [ABSTRACT FROM AUTHOR]

Combining multibranched hybridization chain reaction (mHCR), the photoelectrochemical (PEC) immunosensor was fabricated with a microfluidic paper-based analytical devices using different sizes of CdTe quantum dots (QDs) sensitized flower-like 3D ZnO superstructures as photoactive materials. Firstly, 4-aminothiophenol (PATP) functioned ZnO was anchored on gold-paper working electrode. With the aid of PATP, large-sized CdTe-COOH QDs (QDs1) were conjugated onto the ZnO surface because of the formation of a strong bond (Zn-S) between the thiol of PATP molecule and the ZnO, and the remaining amino group formed an amide bond with carboxylic acid group capping CdTe. Then the small-sized CdTe-NH 2 QDs (QDs2) were modified on the QDs1 by forming amide bond, which leaded to a very strong photocurrent response because of the formation of cosensitized structure. The designed mHCR produced long products with multiple branched arms, which could attached multiple PdAu nanoparticles and catalyze the oxidation of hydroquinone (HQ) using H 2 O 2 as anoxidant. Double strands DNA with multiple branched arms (mdsDNA) was formed by mHCR. In the presence of carcinoembryonic antigen (CEA), PdAu-mdsDNA conjugates-labeled CEA antibody was captured. The concentrations of CEA were measured through the decrease in photocurrent intensity resulting from the increase in steric hindrance of the immunocomplex and the polymeric oxidation product of HQ. In addition, the oxidation product of HQ deposited on the as-obtained electrode, which could efficiently inhibit the photoinduced electron transfer. Under optimal conditions, the PEC immunosensor exhibited excellent analytical performance: the detection range of CEA was from 0.001 to 90 ng mL −1 with low detection limit of 0.33 pg mL −1 . The as-obtained immunosensor exhibited excellent precision, prominent specificity, acceptable stability and reproducibility, and could be used for the detection of CEA in real samples. The proposed assay opens a promising platform of clinical immunoassay for other biomolecules. [ABSTRACT FROM AUTHOR]

Microwire electrodes are presented as an alternative to screen-printed electrodes for detection in electrochemical paper-based analytical devices (ePADs). Compared to carbon ink electrodes, microwire electrodes offer lower resistance and a significant increase in current density relative to carbon ink electrodes. Various microwire compositions and diameters, including 30 μm Pt, 25 μm Au, 18 μm Pt with 8% W, and 15 μm Pt with 20% Ir, were tested and compared to theoretically predicted behavior. The measured current in static solution was below predicted levels for cylindrical microelectrodes but greater than levels predicted for hemi-cylindrical electrodes most likely as a result of the proximity of the electrode to the paper surface. Furthermore, the current response was indicative of semi-thin layer behavior, likely due to the confined solution volume in the paper. After electrode characterization, a device was developed for the non-enzymatic detection of glucose, fructose, and sucrose using a Cu electrode in alkaline solution. The limits of detection for glucose, fructose, and sucrose were 270 nM, 340 nM, and 430 nM, respectively, which are significantly below sugar concentrations found in sweetened beverages or glucose levels in serum. [ABSTRACT FROM AUTHOR]

In this work, we demonstrate a novel low-cost, self-powered paper-based biosensor for glucose monitoring. The device operating mechanism is based on a glucose/oxygen enzymatic fuel cell using an electrochemical energy conversion as a transducing element for glucose monitoring. The self-powered glucose biosensor features (i) a 3D origami paper-based structure for easy system integration onto paper, (ii) an air-cathode on paper for low-cost production and easy operation, and (iii) a screen printed chitosan/glucose oxidase anode for stable current generation as an analytical signal for glucose monitoring. The sensor showed a linear range of output current at 1–5 mM glucose ( R 2 =0.996) with a sensitivity of 0.02 µA mM −1 . The advantages offered by such a device, including a low cost, lack of external power sources/sophisticated external transducers, and the capacity to rapidly generate reliable results, are well suited for the clinical and social settings of the developing world. [ABSTRACT FROM AUTHOR]

Highlights: [•] Develope a semiquantitative method for the detection of glucose with self-calibration based on bienzyme colorimetry by using tree-shaped paper strip. [•] Provide direct outcomes which can be observed by the naked eye or recorded by a simple camera. [•] Judge the level of glucose concentration in blood serum by using Y-shaped paper strip and directly provide a result of “Yes or No”. [•] Provide an affordable, rapid, equipment-free, deliverable and easy-to-use tool for the detection of glucose in human serum sample. [ABSTRACT FROM AUTHOR]

Abstract: Response surface methodology was employed to optimize mixed substrate solid state fermentation for the production of cellulases and xylanase by Aspergillus fumigatus ABK9. Among 11 different parameters, fermentation time (86–88h), medium pH (6.1–6.2), substrate amount (10.0–10.5g) and substrate ratio (wheat bran:rice straw) (1.1) had significantly influences on enzyme production. Under these conditions endoglucanase, β-glucosidase, FPase (filter paper degrading activity) and xylanase activities of 826.2, 255.16, 102.5 and 1130.4U/g, respectively were obtained. The enzyme cocktail extracted (solid to water ratio of 1:10) from the ferments increased brightness of waste office paper pulp by 82.8% ISO, InkD value by 82.1%, removed chromophores (2.53 OD; A237 nm) and hydrophobic compounds (1.15 OD; A465 nm) and also decreased the kappa number to 13.5 from 16.8. [Copyright &y& Elsevier]

Abstract: Ubiquitin activating enzyme (UAE, UBE1, or E1) and seven known homologous “E1s” initiate the conjugation pathways for ubiquitin and 16 other ubiquitin-like modifiers (ULMs) found in humans. The initial step catalyzed by E1s uses adenosine triphosphate (ATP) to adenylate the C terminus of the appropriate ULM and results in the production of inorganic pyrophosphate (PPi). The mechanism of these enzymes can be studied with assays that measure the rate of ULM-dependent ATP:PPi exchange. The traditional method follows the initial velocity of [32P]PPi incorporation into ATP by capturing the nucleotide on activated charcoal powder to separate it from excess [32P]PPi and then measuring [32P]ATP in a scintillation counter. We have modified the method by using charcoal paper to capture the nucleotide and a phosphorimager to quantify the [32P]ATP. The significant increase in throughput that these modifications provide is accomplished without any sacrifice in sensitivity or accuracy compared with the traditional method. To demonstrate this, we reproduce and extend the characterization of the NEDD8 activating enzyme. [Copyright &y& Elsevier]

Abstract: A xylanase gene (xynG1-1) from Paenibacillus campinasensis G1-1 was expressed in Bacillus megaterium MS941 and a high level of extracellular xylansae activity (304.26IU/mL) was achieved after induction with 0.5% xylose. The purified recombinant xylanase (XynG1-1R) revealed optimal activity at 60°C and pH 7.0 and retained 79% and 81% activity after incubation without substrate at 60°C, pH 5.0 and pH 8.0 for 3h, respectively. Application of XynG1-1R (15IU/g pulp) to cotton stalk pulp bleaching increased brightness by 3.65% over that of the control without the xylanase and reduced the need for chlorine compounds by 50% without loss of brightness and pulp fiber qualities. When XynG1-1R (80IU/g paper sludge) was used in combination with mixed cellulolytic enzymes, the saccharification efficiency of recycled paper sludge was increased by 10%. These results indicated that XynG1-1R is a promising candidate for various industrial applications such as biobleaching and bioenergy conversion. [Copyright &y& Elsevier]

Abstract: Two novel cellulase genes, cbh6A and egGH45, were cloned from Phialophora sp. G5 and successfully expressed in Pichia pastoris. The putative polypeptide of CBH6A consists of a family 1 CBM and a catalytic domain of glycosyl hydrolase family 6 cellobiohydrolases, while deduced EgGH45 only contains a catalytic domain of family 45 endoglucanases. CBH6A and EgGH45 were optimally active at pH 7.0 and 65°C, and pH 6.0 and 60°C, respectively. Both enzymes exhibited high activities and stabilities over a wide pH range and had good thermostability at 70°C. CBH6A and EgGH45 had significant resistance to SDS (10mM), remaining 35% and 54% activities, respectively. These enzymes had synergic effect on the hydrolysis of filter paper, showing the highest efficiency in the ratio of CBH6A to EgGH45 at 80:20. The properties make this enzyme combination potential for application in textile and detergents industries. [Copyright &y& Elsevier]

Nearly 100 years ago Michaelis and Menten published their now classic paper [Michaelis, L., and Menten, M. L. (1913) Die Kinetik der Invertinwirkung. Biochem. Z. 49, 333-369] in which they showed that the rate of an enzyme-catalyzed reaction is proportional to the concentration of the enzyme-substrate complex predicted by the Michaelis-Menten equation. Because the original text was written in German yet is often quoted by English-speaking authors, we undertook a complete translation of the 1913 publication, which we provide as Supporting Information. Here we introduce the translation, describe the historical context of the work, and show a new analysis of the original data. In doing so, we uncovered several surprises that reveal an interesting glimpse into the early history of enzymology. In particular, our reanalysis of Michaelis and Menten's data using modern computational methods revealed an unanticipated rigor and precision in the original publication and uncovered a sophisticated, comprehensive analysis that has been overlooked in the century since their work was published. Michaelis and Menten not only analyzed initial velocity measurements but also fit their full time course data to the integrated form of the rate equations, including product inhibition, and derived a single global constant to represent all of their data. That constant was not the Michaelis constant, but rather Vmax/Km, the specificity constant times the enzyme concentration (kcat/Km × E0). [ABSTRACT FROM AUTHOR]

Abstract: Background: Fabry disease is an X-linked lysosomal storage disorder due to deficiency of alpha galactosidase A (AGAL, EC 3.2.1.22). Despite increasing utilization of dried blood spot (DBS) as samples for AGAL enzyme assays, the effects of blood sample collection techniques on enzyme activity have not been studied. Methods: DBS samples were prepared by spotting blood collected into an ethylenediaminetetraacetic acid (EDTA) tube and by direct application of blood from a finger prick or a venipuncture syringe. AGAL activity was measured quantitatively by detecting the fluorescence of 4-methylumbelliferone (4-MU) generated using the substrate 4-methylumbelliferyl-α-d-glucopyranoside (4-MUGal) in an acidic pH for 20 h. N-acetyl-d-galactosamine (GalNAc) was used to inhibit α-galactosidase B (EC 3.2.1.49). Results: We studied 88 previously diagnosed Fabry disease patients and 690 healthy controls. Average AGAL activity in DBS samples prepared using EDTA tubes was higher compared to those spotted directly irrespective of disease status. Conclusions: The study confirms the need for collection method-specific reference ranges using DBS samples. [Copyright &y& Elsevier]

The use of enzymes has been suggested as an environmentally friendly alternative to complement conventional chemical deinking in the recycling of recovered paper. This study compares the use of cellulases/hemicellulases versus the laccase-mediator system for deinking printed fibers from newspapers and magazines. For this purpose, two commercial enzyme preparations with endoglucanase and endoxylanase activities (Viscozyme Wheat from Aspergillus oryzae and Ultraflo L from Humicola insolens, Novozymes) and a commercial laccase (NS51002 from Trametes villosa, Novozymes), the latter in the presence of synthetic or natural (lignin-related) mediators, were evaluated. The enzymatic treatments were studied at the laboratory scale using a standard chemical deinking sequence consisting of a pulping stage; an alkaline stage using NaOH, sodium silicate and fatty acid soap; and a bleaching stage using hydrogen peroxide. The handsheets were then prepared and their brightness, residual ink concentration, and strength properties were measured. Among the different enzymatic treatments assayed, both carbohydrate hydrolases were found to deink the secondary fibers more efficiently. Brightness increased up to 3-4% ISO on newspaper fibers, being Ultraflo 20% more efficient in the ink removal. Up to 2.5% ISO brightness increase was obtained when magazine fibers were used, being Viscozyme 9% more efficient in the ink removal. Regarding the laccase-mediator system, alone or in combination with carbohydrate hydrolases, it was ineffective in deinking both newspaper and magazine fibers, resulting in pulps with worse brightness and residual ink concentration values. However, pulp deinking by the laccase-mediator system was displayed when secondary fibers from printed cardboard were used, obtaining up to 3% ISO brightness increase and lower residual ink concentrations. [ABSTRACT FROM AUTHOR]

The use of artificial zinc finger chimeras to manipulate the expression of a gene of interest is a promising approach because zinc finger proteins can be engineered to bind any given DNA sequence in the genome. We have previously shown that a zinc finger chimera with a VP16 activation domain can activate a reporter gene in transgenic Arabidopsis thaliana (Sánchez, J.P., Ullman, C., Moore, M., Choo, Y. and Chua, N.H. (2002) Regulation of gene expression in Arabidopsis thaliana by artificial zinc finger chimeras. Plant Cell Physiol. 43, 1465–1472). Here, we report the use of artificial zinc finger chimeras to specifically regulate the 4-coumarate:coenzyme-A ligase-1 ( At4CL1) gene in A. thaliana. At4CL1 is a key enzyme in lignin biosynthesis and the down-regulation of At4CL1 can lead to a decrease in lignin content, which has a significant commercial value for the paper industry. To this end, we designed zinc finger chimeras containing either an activation or a repression domain, which bind specifically to the At4CL1 promoter region. Transgenic lines expressing a zinc finger chimera with the VP16 activation domain showed an increase in At4CL1 expression and enzyme activity. In contrast, transgenic lines expressing a chimera with the KOX (KRAB) repression domain displayed repression of At4CL1 expression and enzyme activity. The activation of At4CL1 expression produced an increase in lignin content, and transgenic plant stems showed ectopic lignin distribution. Repression of the At4CL1 gene resulted in reduced lignin content, and lignin distribution in transgenic stems was severely diminished. Our results confirm and extend previous studies of gene regulation using various artificial zinc finger chimeras in animal and plant systems, and show that this system can be used to up- and down-regulate the expression of an endogenous plant gene such as At4CL1. [ABSTRACT FROM AUTHOR]

Selenium was discovered in the first quarter of the 19th century and classified as a chalcogen belonging to the 16th group, along with oxygen, sulfur, tellurium, and polonium. Selenium plays a crucial role in the activation of antioxidant enzymes in the body and helps to reduce oxidative stress by preventing cell damage. It is believed to have cancer-protective effects, including mechanisms such as reducing DNA damage, regulating cell growth, supporting the immune system, and engaging in epigenetic interactions. These are attributed to the antioxidant properties of selenium. The purpose of this paper was to elucidate the effects of selenium exposure on the incidence and mortality of various cancer types using the meta-meta-analysis method. [ABSTRACT FROM AUTHOR]

This paper is a comprehensive review of the techniques for the detection of pancreatic enzymes, which are common biochemical indicators of pancreatitis, including amylase, trypsin, chymotrypsin, elastase, and lipase. Pancreatitis is a disease with self-digestion due to the abnormal activation of digestive enzymes in the pancreas. Hospitalization is often required due to the lack of convenient therapeutic agents. The main recent results are reported in this review, especially the techniques that enable portability and Point-of-Care testing (POCT). This is because timely diagnosis at the early stage and avoiding recurrence after recovery are the keys to treatment. It is also important to reduce the rate of misdiagnosis and to avoid overtreatment. Various detection methods are discussed, with particular attention given to the implementation of chemical sensing and probe design. The new sensing technology for digestive enzymes makes it possible to perform early screening for pancreatitis in remote areas or in one's own home. [ABSTRACT FROM AUTHOR]

Nitrogen (N) and phosphorus (P) additions to forest ecosystems are known to influence various above-ground properties, such as plant productivity and composition, and below-ground properties, such as soil nutrient cycling. However, our understanding of how soil microbial communities and their functions respond to nutrient additions in subtropical plantations is still not complete. In this study, we added N and P to Chinese fir plantations in subtropical China to examine how nutrient additions influenced soil microbial community composition and enzyme activities. The results showed that most soil microbial properties were responsive to N and/or P additions, but responses often varied depending on the nutrient added and the quantity added. For instance, there were more than 30% greater increases in the activities of β-Glucosidase (βG) and N-acetyl-β-D-glucosaminidase (NAG) in the treatments that received nutrient additions compared to the control plot, whereas acid phosphatase (aP) activity was always higher (57 and 71 %, respectively) in the P treatment. N and P additions greatly enhanced the PLFA abundanceespecially in the N2P treatment, the bacterial PLFAs (bacPLFAs), fungal PLFAs (funPLFAs) and actinomycic PLFAs (actPLFAs) were about 2.5, 3 and 4 times higher, respectively, than in the CK. Soil enzyme activities were noticeably higher in November than in July, mainly due to seasonal differences in soil moisture content (SMC). (5G or NAG activities were significantly and positively correlated with microbial PLFAs. There were also significant relationships between gram-positive (G+) bacteria and all three soil enzymes. These findings indicate that G+ bacteria is the most important microbial community in C, N, and P transformations in Chinese fir plantations, and that (βG and NAG would be useful tools for assessing the biogeochemical transformation and metabolic activity of soil microbes. We recommend combined additions of N and P fertilizer to promote soil fertility and microbial activity in this kind of plantation. [ABSTRACT FROM AUTHOR]

High pressure processing (HPP) offers the benefits of safety, uniformity, energy-efficient, and low waste, which is widely applied for microbial inactivation and shelf-life extension for foods. Over the past forty years, HPP has been extensively researched in the food industry, enabling the inactivation or activation of different enzymes in future food by altering their molecular structure and active site conformation. Such activation or inactivation of enzymes effectively hinders the spoilage of food and the production of beneficial substances, which is crucial for improving food quality. This paper reviews the mechanism in which high pressure affects the stability and activity of enzymes, concludes the roles of key enzymes in the future food processed using high pressure technologies. Moreover, we discuss the application of modified enzymes based on high pressure, providing insights into the future direction of enzyme evolution under complex food processing conditions (e.g. high temperature, high pressure, high shear, and multiple elements). Finally, we conclude with prospects of high pressure technology and research directions in the future. Although HPP has shown positive effects in improving the future food quality, there is still a pressing need to develop new and effective combined processing methods, upgrade processing modes, and promote sustainable lifestyles. [ABSTRACT FROM AUTHOR]

Dynamic constitutional materials are constructed from reversibly connecting components adaptively self-assembling under the pressure of internal or external factors. Functional macromonomeric poly-(ethylene glycol) segments and 3D connecting cores can be used to conceive biomimetic dynamic constitutional frameworks (DCFs) for selective/adaptive biomolecular encapsulation of proteins, enzymes, etc. Gold nanoparticles (AuNps) can be involved as core connectors to expand the networks of DCFs at the nanoscale, thereby increasing the biomolecular encapsulation and activity. Efficient encapsulation of bovine carbonic anhydrase (bCA) in host matrixes is of tremendous importance for maintaining its stability, catalytic activity, and durability. In this paper, we demonstrate that AuNps-DCFs-bCA bioconjugates, synthesized via reversible imine/amino-carbonyl as sulfide-AuNps chemistries, assemble into nanometric aggregates of around 200 nm with high stability for bCA encapsulation. This system showed an impressive improved catalytic activity, more than 30 times higher than that of bCA alone. The Km values were two to three times lower compared to free bCA indicating a stronger affinity for bCA under encapsulation, while Vmax was higher. This performance is very promising for current state-of-the-art carbon dioxide capture systems with equivalent amounts of enzyme, even after heating for a prolonged period at 100 °C, translating into a direct perspective for enhancing flue gas capture and conversion performances. [ABSTRACT FROM AUTHOR]

Microbes are major players in leaf litter decomposition and therefore advances in the understanding of their control on element cycling are of paramount importance. Our aim was to investigate the influence of leaf litter stoichiometry in terms of carbon (C): nitrogen (N): phosphorus (P) on the decomposition process, and to follow changes in microbial community structure and function in response to temperature-stress treatments. To elucidate how the stoichiometry of beech litter (Fagus sylvatica L.) and stress treatments interactively affect the decomposition processes, a terrestrial microcosm experiment was conducted. Beech litter from different Austrian sites covering C:N ratios from 39 to 61 and C:P ratios from 666 to 1729 were incubated at 15 °C and 60% moisture for six months. Part of the microcosms were then subjected to severe changes in temperature (+30 °C and -15 °C) to monitor the influence of temperature stress. Extracellular enzyme activities were assayed and respiratory activities measured. A semi-quantitative metaproteomics approach (1D-SDS PAGE combined with liquid chromatography and tandem mass-spectrometry; unique spectral counting) was employed to investigate the impact of the applied stress treatments in dependency of litter stoichiometry on structure and function of the decomposing community. In litter with narrow C:nutrient ratios microbial decomposers were most abundant. Cellulase, chitinase, phosphatase and protease activity decreased after heat and frost treatments. Decomposer communities and specific functions varied with site i.e. stoichiometry. The applied stress evoked strong changes of enzyme activities, dissolved organic nitrogen and litter pH. Freeze treatments resulted in a decline in residual plant litter material, and increased fungal abundance indicating slightly accelerated decomposition. Overall, we could detect a strong effect of litter stoichiometry on microbial community structure as well as function. Temperature perturbations resulted in short- to medium-term alterations of microbial function, especially heat blocked decomposing enzymes. [ABSTRACT FROM AUTHOR]

Two fungi (unidentified) were isolated from soil and marine environments. These isolates were used for bioremediation of pulp and paper mill effluent at the laboratory scale. The treatment resulted in the reduction of color, lignin, and COD of the effluent in the order of 78.6%, 79.0%, and 89.4% in 21 days. A major part of reductions in these parameters occurred within 5 days of the treatment, which was also characterized by a steep decline in the pH of the effluent. The enzyme activity of these fungi was also tested, and the clearance zone was obtained in the plate assay. [ABSTRACT FROM AUTHOR]

Cytochrome P450s (Cyt P450s) and peroxidases are enzymes featuring iron heme cofactors that have wide applicability as biocatalysts in chemical syntheses. Cyt P450s are a family of monooxygenases that oxidize fatty acids, steroids, and xenobiotics, synthesize hormones, and convert drugs and other chemicals to metabolites. Peroxidases are involved in breaking down hydrogen peroxide and can oxidize organic compounds during this process. Both heme-containing enzymes utilize active FeIV＝O intermediates to oxidize reactants. By incorporating these enzymes in stable thin films on electrodes, Cyt P450s and peroxidases can accept electrons from an electrode, albeit by different mechanisms, and catalyze organic transformations in a feasible and cost-effective way. This is an advantageous approach, often called bioelectrocatalysis, compared to their biological pathways in solution that require expensive biochemical reductants such as NADPH or additional enzymes to recycle NADPH for Cyt P450s. Bioelectrocatalysis also serves as an ex situ platform to investigate metabolism of drugs and bio-relevant chemicals. In this paper we review biocatalytic electrochemical reactions using Cyt P450s including C–H activation, S-oxidation, epoxidation, N-hydroxylation, and oxidative N-, and O-dealkylation; as well as reactions catalyzed by peroxidases including synthetically important oxidations of organic compounds. Design aspects of these bioelectrocatalytic reactions are presented and discussed, including enzyme film formation on electrodes, temperature, pH, solvents, and activation of the enzymes. Finally, we discuss challenges and future perspective of these two important bioelectrocatalytic systems. [ABSTRACT FROM AUTHOR]

Severe acute pancreatitis (SAP) begins with premature activation of enzymes, promoted by the immune system, triggering a potential systemic inflammatory response that leads to organ failure with increased mortality and a bleak prognosis. Interleukin-22 (IL-22) is a cytokine that may have a significant role in SAP. IL-22, a member of the IL-10 cytokine family, has garnered growing interest owing to its potential tissue-protective properties. Recently, emerging research has revealed its specific effects on pancreatic diseases, particularly SAP. This paper provides a review of the latest knowledge on the role of IL-22 and its viability as a therapeutic target in SAP. Highlights: 1. Interleukin-22 (IL-22) is a member of the IL-10 family of cytokines. 2. IL-22 is a potential indicator for monitoring organ dysfunction and severity of severe acute pancreatitis (SAP). 3. IL-22 acts as a protective factor against acinar cell damage by suppressing autophagy and reducing the infiltration of inflammatory cells. 4. IL-22 alleviate SAP-induced multiorgan injury. 5. IL-22 protect SAP-induced intestinal barrier damage. [ABSTRACT FROM AUTHOR]

Abstract: To produce bioactive papers, i.e. papers incorporating biomolecules that are useful for analyte detection, adequate immobilization strategies should be devised. In this article, the physical immobilization behavior and activity of the enzyme horseradish peroxidase (HRP) on various papermaking substrates were studied. The papermaking substrates included amorphous and crystalline cellulose, calcium carbonate, styrene butadiene latex, polystyrene, and both negatively charged rayon and rayon with a positively charged layer. It was found that HRP adsorption improves as the hydrophobicity of the substrate increases; however, excessive hydrophobicity produces enzyme deactivation. HRP–calcium carbonate binding was weak and the enzyme loading was scant. These results provided a possible explanation for the poor analytical signals observed in pigment-coated papers when used as bioactive paper supports. Electrostatic effects played a minor role in HRP adsorption behavior. [Copyright &y& Elsevier]

Endo-ß-1,4-glucanase AkEG21 belonging to glycosyl hydrolase family 45 (GHF45) is the most abundant cellulase in the digestive fluid of sea hare (Aplysia kurodai). The specific activity of this 21-kDa enzyme is considerably lower than those of other endo ß-1,4-glucanases in the digestive fluid of A. kurodai, therefore its role in whole cellulose hydrolysis by sea hare is still uncertain. Although AkEG21 has a catalytic domain without a cellulose binding domain, it demonstrated stable binding to cellulose fibers, similar to that of fungal cellobiohydrolase (CBH) 1 and CBH 2, which is strongly inhibited by cellohexaose, suggesting the involvement of the catalytic site in cellulose binding. Cellulose-bound AkEG21 hydrolyzed cellulose to cellobiose, cellotriose and cellotetraose, but could not digest an external substrate, azo-carboxymethyl cellulose. Cellulose hydrolysis was considerably stimulated by the synergistic action of cellulose-bound AkEG21 and AkEG45, another ß-1,4-endoglucanase present in the digestive fluid of sea hare; however no synergy in carboxymethylcellulose hydrolysis was observed. When AkEG21 was removed from the digestive fluid by immunoprecipitation, the cellulose hydrolyzing activity of the fluid was significantly reduced, indicating a critical role of AkEG21 in cellulose hydrolysis by A. kurodai. These findings suggest that AkEG21 is a processive endoglucanase functionally equivalent to the CBH, which provides a CBH-independent mechanism for the mollusk to digest seaweed cellulose to glucose. [ABSTRACT FROM AUTHOR]

A comprehensive density functional theory investigation has been presented towards the comparison of the C–H activation reactivity between high-valent iron-oxo and ruthenium-oxo complexes. A total of four compounds, e.g., [Ru(IV)O(tpy-dcbpy)] (1), [Fe(IV)O(tpy-dcbpy)] (1′), [Ru(IV)O(TMCS)] (2), and [Fe(IV)O(TMCS)] (2′), have been considered for this investigation. The macrocyclic ligand framework tpy(dcbpy) implies tpy = 2,2′:6′,2′′-terpyridine, dcbpy = 5,5′-dicarboxy-2,2′-bipyridine, and TMCS is TMC with an axially tethered –SCH2CH2 group. Compounds 1 and 2′ are experimentally synthesized standard complexes with Ru and Fe, whereas compounds 1′ and 2 were considered to keep the macrocycle intact when switching the central metal atom. Three reactants including benzyl alcohol, ethyl benzene, and dihydroanthracene were selected as substrates for C–H activation. It is noteworthy to mention that Fe(IV)O complexes exhibit higher reactivity than those of their Ru(IV)O counterparts. Furthermore, regardless of the central metal, the complex featuring a tpy-dcbpy macrocycle demonstrates higher reactivity than that of TMCS. Here, a thorough analysis of the reactivity-controlling characteristics—such as spin state, steric factor, distortion energy, energy of the electron acceptor orbital, and quantum mechanical tunneling—was conducted. Fe(IV)O exhibits the exchanged enhanced two-state-reactivity with the quintet reactive state, whereas Ru(IV)O has only a triplet reactive state. Both the distortion energy and acceptor orbital energy are low in the case of Fe(IV)O supporting its higher reactivity. All the investigated C–H activation processes involve a significant contribution from hydrogen tunneling, which is more pronounced in the case of Ru, although it cannot alter the reactivity pattern. Furthermore, it has also been found that, independent of the central metal, aliphatic hydroxylation is always preferable to aromatic hydroxylation. Overall, this work is successful in establishing and investigating the cause of enzymes' natural preference for Fe over Ru as a cofactor for C–H activation enzymes. [ABSTRACT FROM AUTHOR]

Many chronic diseases such as Alzheimer's disease, diabetes, and cardiovascular diseases are closely related to in vivo oxidative stress caused by excessive reactive oxygen species (ROS). Natural polysaccharides, as a kind of biomacromolecule with good biocompatibility, have been widely used in biomedical and medicinal applications due to their superior antioxidant properties. In this review, scientometric analysis of the highly cited papers in the Web of Science (WOS) database finds that antioxidant activity is the most widely studied and popular among pharmacological effects of natural polysaccharides. The antioxidant mechanisms of natural polysaccharides mainly contain the regulation of signal transduction pathways, the activation of enzymes, and the scavenging of free radicals. We continuously discuss the antioxidant activities of natural polysaccharides and their derivatives. At the same time, we summarize their applications in the field of pharmaceutics/drug delivery, tissue engineering, and antimicrobial food additives/packaging materials. Overall, this review provides up-to-date information for the further development and application of natural polysaccharides with antioxidant activities. [ABSTRACT FROM AUTHOR]

The enormous quantity of plant waste produced from greenhouse tomato crops is an environmental problem that should be solved by recycling that waste into valuable organic products through low-cost technologies, such as vermicomposting. Feasibility of vermicomposting greenhouse tomato-plant waste (P) using paper-mill sludge (S) as complementary waste was investigated by this study. Earthworm development in P, S, and two mixtures of both wastes was monitored over 24 weeks and compared with that in cow dung (D), an optimum organic-waste to be vermicomposted. The effectiveness of vermicomposting to biostabilize those wastes was assessed by analysing phospholipid fatty acid composition, chemical features, plant-nutrient content, metal concentration, enzyme activities, and germination index (GI). A commercial vermicompost was also analyzed and taken as a reference of vermicompost quality. Earthworms did not survive in P alone, but a mixture of P with S at a ratio of 2:1 or 1:1 resulted in earthworm development similar to that observed in D. Phospholipid fatty acid analysis revealed that earthworm activity strongly transformed initial microbiota inhabiting the wastes, giving rise to vermicompost microbial communities which were similar to that of a commercial vermicompost. Both mixtures of P and S were properly biostabilized through vermicomposting, as indicated by decreases in their C:N ratio and enzyme activities together with increases in their degree of maturity (GI ~ 100 %) after the process. This study demonstrates that the vermicomposting of tomato-plant waste together with paper-mill sludge allows the recycling of both wastes, thereby improving the environmental sustainability of greenhouse crops. [ABSTRACT FROM AUTHOR]

Crude Manganese Peroxidase (MnP) preparations from the fungus Bjerkandera sp. BOS55 were tested for their abilities to bleach and delignify oxygen delignified Eucalyptus Kraft pulp in a sequence combining a chelating and a peroxide stage. The inclusion of the enzymatic treatment in the bleaching sequence helped considerably to improve the final brightness. Other interesting results included moderate Kappa reduction, slight yield loss and paper sheets with an acceptable breaking length and burst index. [ABSTRACT FROM AUTHOR]

Nanobodies and chemical cross-linking were used to gain information on the identity and positions of flexible domains of PI3Kα. The application of chemical cross-linking mass spectrometry (CXMS) facilitated the identification of the p85 domains BH, cSH2, and SH3 as well as their docking positions on the PI3Kα catalytic core. Binding of individual nanobodies to PI3Kα induced activation or inhibition of enzyme activity and caused conformational changes that could be correlated with enzyme function. Binding of nanobody Nb3-126 to the BH domain of p85α substantially improved resolution for parts of the PI3Kα complex, and binding of nanobody Nb3-159 induced a conformation of PI3Kα that is distinct from known PI3Kα structures. The analysis of CXMS data also provided mechanistic insights into the molecular underpinning of the flexibility of PI3Kα. [ABSTRACT FROM AUTHOR]

This paper describes the synthesis and characterization of new bivalent folate-targeted PEGylated doxorubicin (FA2-dPEG-DOX2) made by modular chemo-enzymatic processes using Candida antarctica lipase B (CALB) as a biocatalyst. Unique features are the use of monodisperse PEG (dPEG) and the synthesis of thiol-functionalized folic acid yielding exclusive γ-conjugation of folic acid (FA) to dPEG. The polymer-based drug conjugate is built up by a series of transesterification and Michael addition reactions all catalyzed be CALB. In comparison with other methods in the literature, the modular approach with enzyme catalysis leads to selectivity, full conversion and high yield, and no transition metal catalyst residues. The intermediate product with four acrylate groups is an excellent platform for Michael-addition-type reactions for a wide variety of biologically active molecules. The chemical structures were confirmed by nuclear magnetic resonance spectroscopy (NMR). Flow cytometry analysis showed that, at 10 µM concentration, both free DOX and FA2-dPEG-DOX2 were taken up by 99.9% of triple-negative breast cancer cells in 2 h. Fluorescence was detected for 5 days after injecting compound IV into mice. Preliminary results showed that intra-tumoral injection seemed to delay tumor growth more than intravenous delivery. [ABSTRACT FROM AUTHOR]

In the present study, thermophilic fungus Thermoascus aurantiacus, a local isolate, was used for production of cellulolytic enzymes using rice straw (RS) as substrate under submerged fermentation using Box-Behnken Design (BBD) of Response Surface Methodology (RSM). Themophilic fungus T. aurantiacus grew very well at 50-55°C temperature. BBD was used to study the influence of process parameters, pH (4-6), temperature (40-60°C) and substrate concentration (2-8 g/L) and their interactive effect on cellulase enzyme production. The R2 value 0.94 for Filter paper activity and 0.95 for carboxymethyl cellulase activity indicate that the model is appropriate and replicated to predict the effect of pH, temperature and substrate concentration on both enzyme activities. Under optimized conditions, 6.1 (U/gds) FPase activity and 28.2 (U/gds) CMCase activity was observed. [ABSTRACT FROM AUTHOR]

Excess of sludge generated during activated sludge process by various industries is causing sludge disposable problem worldwide. Sludge contains organic materials of a highly complex mixture of molecules with some toxic compounds. Thus, the harmful effect of sludge could be managed by processing of sludge and recovery of useful products. Sludge could undergo biodegradation and used for the recovery of enzymes and energy. Enzymes (amylase, alkaline phosphatase, lipase, proteases) are produced by microorganisms and released into the media to degrade organic matters of sludge. These enzymes can be extracted by means of different physical and chemical methods from activated sludge for the multifarious application. Current manuscript discussed all the methods utilized for the extraction of enzyme and their utilization in terms of energy generation. Extracted industrial enzymes are used in agriculture, dairy, detergents, pulp, paper, cosmetics, and pharmaceutics. Apart from enzyme extraction, production of energy (biofuels, electricity) is also done by key treatment of sludge. It is a compilation of current knowledge and expected competitiveness with respect to existing methods already applied in practice for enzyme recovery. This paper presents an overview of the production of valuable enzymes and various forms of energy from sludge, which is toxic and unwanted for life. [ABSTRACT FROM AUTHOR]

Aim. Currently available medicines have little to offer in terms of supporting the regeneration of injured hepatic cells. Previous experimental studies have shown that resveratrol and metformin, less specific activators of AMP-activated protein kinase (AMPK) and sirtuin 1 (SIRT1), can effectively attenuate acute liver injury. The aim of this experimental study was to elucidate whether modulation of AMPK and SIRT1 activity can modify drug/paracetamol (APAP)-induced hepatocyte damage in vitro. Methods. Primary rat hepatocytes were pretreated with mutual combinations of specific synthetic activators and inhibitors of SIRT1 and AMPK and followed by a toxic dose of APAP. At the end of cultivation, medium samples were collected for biochemical analysis of alanine-aminotransferase and nitrite levels. Hepatocyte viability, thiobarbituric reactive substances, SIRT1 and AMPK activity and protein expression were also assessed. Results. The harmful effect of APAP was associated with decreased AMPK and SIRT1 activity and protein expression alongside enhanced oxidative stress in hepatocytes. The addition of AMPK activator (AICAR) or SIRT1 activator (CAY10591) significantly attenuated the deleterious effects of AMPK inhibitor (Compound C) on the hepatotoxicity of APAP. Furthermore, CAY10591 but not AICAR markedly decreased the deleterious effect of APAP in combination with SIRT1 inhibitor (EX-527). Conclusion. Our findings demonstrate that decreased AMPK activity is associated with the hepatotoxic effect of APAP which can be significantly attenuated by the administration of a SIRT1 activator. These findings suggest that differentiated modulation of AMPK and SIRT1 activity could therefore provide an interesting and novel therapeutic opportunity in the future to combat hepatocyte injury.

Glycosyltransferases (GTs) are carbohydrate-active enzymes (CAZy) involved in the synthesis of natural glycan structures. The application of CAZy is highly demanded in biotechnology and pharmaceutics. However, it is being hindered by the lack of high-quality and comprehensive repositories of the research data accumulated so far. In this paper, we describe a new curated Carbohydrate Structure Glycosyltransferase Database (CSDB_GT). Currently, CSDB_GT provides ca. 780 activities exhibited by GTs, as well as several other CAZy, found in Arabidopsis thaliana and described in ca. 180 publications. It covers most published data on A. thaliana GTs with evidenced functions. CSDB_GT is linked to the Carbohydrate Structure Database (CSDB), which stores data on archaeal, bacterial, fungal and plant glycans. The CSDB_GT data are supported by experimental evidences and can be traced to original publications. CSDB_GT is freely available at http://csdb.glycoscience.ru/gt.html. [ABSTRACT FROM AUTHOR]

Enhancing cellulase recyclability is a promising approach to reducing the high enzyme cost for activation of dissolving pulp. For this purpose, Tween 80, an amphiphilic surfactant, was employed to facilitate cellulase recovery from fibers. Results indicated that the Cellulase–Tween complex was formed (with a particle size increase of 140%) through a hydrophobic interaction. A much lower maximum binding capacity and affinity (i.e., absorption ability) of cellulase onto fibers (6.03 versus 24.3 mg protein/g cellulose and 7.45 versus 13.8 ml/mg protein) were achieved for the complex in comparison with the control. The recovered cellulase activity was increased up to 65.7% from the control one in five consecutive recycle rounds with Tween 80 addition. As a result, the recovered cellulase efficiency was improved in terms of viscosity and reactivity. Other properties, such as α-cellulose content, alkaline solubility, molecular weight distribution, morphology, and crystalline structure, were all supported by the positive effect of Tween 80 in cellulase recycling. [ABSTRACT FROM AUTHOR]

Background: Abnormalities in glycan biosynthesis have been conclusively related to various diseases, whereas the complexity of the glycosylation process has impeded the quantitative analysis of biochemical experimental data for the identification of glycoforms contributing to disease. To overcome this limitation, the automatic construction of glycosylation reaction networks in silico is a critical step. Results: In this paper, a framework K2014 is developed to automatically construct N-glycosylation networks in MATLAB with the involvement of the 27 most-known enzyme reaction rules of 22 enzymes, as an extension of previous model KB2005. A toolbox named Glycosylation Network Analysis Toolbox (GNAT) is applied to define network properties systematically, including linkages, stereochemical specificity and reaction conditions of enzymes. Our network shows a strong ability to predict a wider range of glycans produced by the enzymes encountered in the Golgi Apparatus in human cell expression systems. Conclusions: Our results demonstrate a better understanding of the underlying glycosylation process and the potential of systems glycobiology tools for analyzing conventional biochemical or mass spectrometry-based experimental data quantitatively in a more realistic and practical way. [ABSTRACT FROM AUTHOR]

[Display omitted] • In situ FZC honeycomb spheres for cellulase direct adsorption and immobilization. • ZIF-8 components of the carrier regulate secondary structure of cellulase. • Thermal activation of immobilized enzyme was reported. • Application of immobilized cellulase in biomass saccharification. This paper presents a promising heterogeneous biocatalyst-immobilized cellulase system, which can facilitate the targeted catalytic conversion of biomass to reducing sugars. Magnetic Zeolite Imidazole Framework-8/chitosan (Fe 3 O 4 /ZIF-8/CS, FZC) hybrid honeycomb microspheres were synthesized to immobilize cellulase using an in-situ growth strategy. The enhancement of immobilization efficiency induced by each constituent of the composite carrier was explored. This exploration was grounded in varying combinations of components within the honeycomb composite spheres, revealing the indispensability of ZIF-8. Through in-situ assembly, the composite material presents a microsphere with an internal honeycomb structure and multilevel pores. Effective immobilization, temperature-responsive conformational change, and cyclic stability were achieved due to the scaling effect of the channel, extensive specific surface area, and abundant amino group interactions enable immobilized enzymes to maintain excellent performance under challenging operating conditions, demonstrating an enhanced structure-performance relationship. With the obtained immobilized cellulase, we achieved a high load (461.67 mg/g) of cellulase and a significant reducing sugar yield (328.39 mg/g) from corncob waste. In addition, we investigated the enhancement effect of thermal activation on the immobilized enzyme activity of composite carrier. The results showed that the appropriate heat treatment was conducive to the activation of FZC immobilized enzyme with a reducing sugar yield 1.61 times higher than that of the control. The strategy provided in this study promotes the development of immobilized biocatalysts and provides a useful reference for the further design of biocatalytic systems. [ABSTRACT FROM AUTHOR]

Macrophages represent primary players of the innate immune system. Macrophage activation triggers several signaling pathways and is tightly associated with metabolic changes, which drive different immune subsets. Recent studies unveil the role of various metabolic enzymes in macrophage activation. Here, we show that malic enzyme 1 (ME1) is overexpressed in LPS-induced macrophages. Through chromatin immunoprecipitation, we demonstrate that ME1 transcriptional regulation is under control of NF-κB. Furthermore, ME1 activity is also increased in activated human PBMC-derived macrophages. Notably, ME1 gene silencing decreases nitric oxide as well as reactive oxygen species and prostaglandin E2 inflammatory mediators. Therefore, modulating ME1 provides a potential approach for immunometabolic regulation and in turn macrophage function. [ABSTRACT FROM AUTHOR]

The Sogatella furcifera (Horva'th) (Homoptera: Delphacidae) is a white-backed planthopper (WBPH) that causes "hopper burn" in rice, resulting in severe yield loss. Gamma-aminobutyric acid (GABA) is a well-known neurotransmitter that inhibits neurotransmission in insects by binding to specific receptors. In this study, we investigated the potential role of GABA in modulating rice resistance to WBPH and evaluated possible defense mechanisms. The experiment was conducted in green house in pots consist of four groups: control, GABAtreated, WBPH-infested, and WBPH-infested treated with GABA. Among the various tested concentration of GABA, 15 mM GABA was applied as a single treatment in water. The treatment was administered one week before WBPH infestation. The results revealed that 15 mM GABA treatment strongly increased WBPH resistance. A plate-based assay indicated that direct application of 15 mM GABA increased the mortality rate of WBPH and increased the damage recovery rate in rice plants. We found that GABA treatment increased the activation of antioxidant enzymes and reduced the reactive oxygen species content and malondialdehyde contents, and reduced the damage rate caused by WBPH. Interestingly, GABA-supplemented plants infested with WBPH exhibited increased phenylalanine ammonia-lyase and pathogenesis-related (PR) genes expression levels. GABA induced the accumulation of abscisic acid (ABA) and salicylic acid (SA) and enhanced the stomata closure and reduced leaf vessels to reduce water conductance during WBPH stress. Furthermore, we found that GABA application to the plant induced the expression of Jasmonic acid (JA) biosynthesis genes (LOX, AOS, AOC, and OPR) and melatonin biosynthesisrelated genes (TDC, T5H, ASMT, and SNAT). Our study suggested that GABA increases resistance against WBPH infestation by regulating antioxidant defense system, TCA cycle regulation, phytohormonal signaling, and PR gene regulation. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus signal peptidase IB (SpsB) is an essential enzyme for protein secretion. While inhibition of its activity by small molecules is a well-precedented mechanism to kill bacteria, the mode of activation is however less understood. We here investigate the activation mechanism of a recently introduced activator, the antibiotic compound PK150, and demonstrate by combined experimental and Molecular Dynamics (MD) simulation studies a unique principle of enzyme stimulation. Mass spectrometric studies with an affinity-based probe of PK150 unravel the binding site of PK150 in SpsB which is used as a starting point for MD simulations. Our model shows the localization of the molecule in an allosteric pocket next to the active site which shields the catalytic dyad from excess water that destabilizes the catalytic geometry. This mechanism is validated by the placement of mutations aligning the binding pocket of PK150. While the mutants retain turnover of the SpsB substrate, no stimulation of activity is observed upon PK150 addition. Overall, our study elucidates a previously little investigated mechanism of enzyme activation and serves as a starting point for the development of future enzyme activators. The study explores the activation mechanism of Staphylococcus aureus signal peptidase IB by the antibiotic PK150. We identified that PK150 shields the active site from excess water and stabilizes its catalytic geometry, thus enhancing catalysis. [ABSTRACT FROM AUTHOR]

Substrate impregnated paper discs were prepared using peptidyl derivatives of 7-amino-4-trfluoromethylcoumarin (AFC). After incubation with test solutions. the green. UV-induced fluorescence of AFC liberated by enzyme activity was distinguishable from the blue-vioIet fluorescence of the substrates. The AEC could then be coupled with p-dimethylaminocinnamaldehyde to form a colored Schiff base Semi-quantitatise assessments of disc fluorescence and color were made by comparison with AFC substrate standards Assays with discs impregnated with MeOSuc-Ala-Ala-Pro-Val-AFC, Z-Gly-Gly-Arg-AFC and Ala-Pro-AFC for elastase-, trypsin-. and dipeplidyl peptidase (DPP) IV like activities respectively were evaluated using purified DPP IV and 100 eluates of crevicular fluid collected on filter paper strips from 10 gingivitis and periodontitis patients .The results showed that, within their working ranges. scores of disc fluorescence and color were reasonably accurate and reliable by comparison with enzyme activities measured in parallel quantitative fluorimetric assays with the same substrates Using disc color which was more sensitive than fluorescence, it was generally possible to measure all three enzyme activities in crevicular fluid samples from 5 periodontitis patients with varying degrees of gingival inflammation and pocketing Disc color assays require no special apparatus and could be used for enzyme estimations in the clinical setting. [ABSTRACT FROM AUTHOR]

To enhance recovery yield of carboxymethylcellulase (CMCase), E. coli BL21/A-53 producing the histidine-tagged CMCase was constructed in this study. The recovery yield of the histidine-tagged CMCase using the His-tag affinity chromatography was 39.8%. The predicted molecular weight of the histidine-tagged CMCase was determined as 56,260 Da. Its Km and Vmax were 9.3 g l−1 and 76.3 g l−1·min−1, respectively. The histidine-tagged CMCase hydrolyzed avicel, carboxymethylcellulose (CMC), filter paper, pullulan, xylan, but there was no detectable activity on cellobiose, p-Nitrophenyl-β-D-glucopyranoside (pNPG). The optimal temperature and pH for the enzymatic reaction of the histidine-tagged CMCase was 50 °C and 5.0. The histidine-tagged CMCase was enhanced by CoCl2 until the concentration of 100 mM, but inhibited by EDTA, HgCl2, MnCl2, NiCl2, and RbCl2. The characteristics of the histidine-tagged CMCase produced by E. coli BL21/A-53 were compared with those of CMCase without the histidine-tag of Bacillus subtilis subsp. subtilis A-53. The little changed characteristics of the histidine-tagged CMCase compared to the CMCase without a His-tag seemed to be the conformational change in the structure due to a His-tag. [ABSTRACT FROM AUTHOR]

Abstract Progressing towards sustainable development remains a key global challenge. And yet, the various interpretations of the concept of sustainable development and the questions it raises about economic growth make its implementation difficult. Higher education institutions may help to overcome these difficulties by developing new processes of change. However, to achieve this they need to integrate sustainable development in all their areas of activity. The aim of this paper was to develop new insights into organisational change processes in universities relating to sustainable development. Contributing to this aim, this paper reports on a case study of United Kingdom higher education drawing on findings and conclusions from a survey of their policy frameworks relating to sustainable development. The method comprised a critical policy analysis in order to identify, differentiate and categorise stakeholder interactions. The data generated comprised the range of higher education stakeholders and the network of interactions that they formed. Theoretical insights from social network analysis, stakeholder theory and the normative business model were used to find opportunities to address the difficulties in the implementation of sustainable development. Results suggested that the existing networks identified in the policy frameworks may not support the effective integration of sustainable development in higher education. Low-density of the national networks; the lack of a clear governance vocabulary for national policy frameworks; and the lack of explicit funding flows between organisations all pose problems for organisational change towards sustainable development in higher education. Highlights • The paper develops insights in organisational change at universities in sustainable development. • Critical policy analysis identifies, differentiates and categorises stakeholder interactions. • Social network analysis, stakeholder theory, Normative Business Model inform interpretation. • Density, governance and funding of policy networks could aid change at universities. [ABSTRACT FROM AUTHOR]

The lytic polysaccharide monooxygenases (LPMOs) are a group of enzymes that have been extensively studied due to their ability to potentiate the deconstruction of cellulosic biomass to glucose as a feedstock for fermentation processes. On the other hand, the unique ability of LPMOs to introduce carbonyl groups (C4-keto or C1-carboxylate groups) at cleavage points in cellulose chains presents an opportunity for further chemical functionalisation using orthogonally reactive handles. Yet, enzyme reaction parameters likely require careful control to balance maximum functional group incorporation versus fibre erosion by lytic cleavage to soluble oligosaccharides. In this study, we explored the activity of a representative bacterial C1-oxidising cellulolytic LPMO, Cellulomonas flavigena LPMO10A (CflaLPMO10A), for the controlled introduction of reactive carboxylate groups on a range of exemplar wood pulp fibres, cellulose nanocrystals (CNC) derived from wood pulp, and bacterial cellulose. We optimized enzyme loading as a function of cellulose mass using HPLC analysis of soluble oligosaccharide products and conductometric titration of surface carboxylic acid groups as readouts of cellulose degradation and modification, respectively. (2,2,6,6-Tetramethylpiperidin-1-yl)oxyl (TEMPO)-mediated C6 oxidation served as a benchmark chemical reaction. Gratifyingly, CflaLPMO10A was effective for the controlled carboxylation of essentially all of the selected cellulosic substrates, with the exception of highly sulphated CNCs. Controlled LPMO oxidation generally did not alter the morphology of pulp fibres nor CNC, except in the case of mechanically sheared pulps, which evidenced surface smoothening. Furthermore, the resulting carboxylated materials were amenable to subsequent chemical modification under mild conditions, as demonstrated by the conjugation of fluorescent dyes and nanoparticles as examples. Our results highlight the potential of LPMOs as orthogonal tools for the controlled, chemo-enzymatic modification of cellulosic materials for advanced applications. [ABSTRACT FROM AUTHOR]

A light-driven deazaflavin-dependent direct enzyme regeneration system has been developed for a P450-BM3 catalyzed CH-activating hydroxylation, thereby avoiding the need for the expensive NADPH cofactor. [ABSTRACT FROM AUTHOR]

A simple and novel gold nanoparticle (Au-NPs) based colorimetric method has been developed for efficient screening of class A β-lactamase (Bla) activity and inhibitors in vitroand in bacterial strains. [ABSTRACT FROM AUTHOR]