Bioactivation of Konjac Glucomannan Films by Tannic Acid and Gluconolactone Addition.

Wound healing is a dynamic process that requires an optimal extracellular environment, as well as an accurate synchronization between various cell types. Over the past few years, great efforts have been devoted to developing novel approaches for treating and managing burn injuries, sepsis, and chronic or accidental skin injuries. Multifunctional smart-polymer-based dressings represent a promising approach to support natural healing and address several problems plaguing partially healed injuries, including severe inflammation, scarring, and wound infection. Naturally derived compounds offer unique advantages such as minimal toxicity, cost-effectiveness, and outstanding biocompatibility along with potential anti-inflammatory and antimicrobial activity. Herein, the main driving idea of the work was the design and development of konjac glucomannan d-glucono-1,5-lactone (KG) films bioactivated by tannic acid and d-glucono-1,5-lactone (GL) addition. Our analysis, using attenuated total reflectance-Fourier transform infrared, atomic force microscopy, and surface energy measurements demonstrated that tannic acid (TA) clearly interacted with the KG matrix, acting as its cross-linker, whereas GL was embedded within the polymer structure. All developed films maintained a moist environment, which represents a pivotal property for wound dressing. Hemocompatibility experiments showed that all tested films exhibited no hemolytic impact on human erythrocytes. Moreover, the presence of TA and GL enhanced the metabolic and energetic activity in human dermal fibroblasts, as indicated by the MTT assay, showing results exceeding 150%. Finally, all films demonstrated high antibacterial properties as they significantly reduced the multiplication rate of both Staphylococcus aureus and Escherichia coli in bacterial broth and created the inhibition zones for S. aureus in agar plates. These remarkable outcomes make the KG/TA/GL film promising candidates for wound healing applications.

6-Phosphogluconolactonase is critical for the efficient functioning of the pentose phosphate pathway.

The metabolic networks of microorganisms are remarkably robust to genetic and environmental perturbations. This robustness stems from redundancies such as gene duplications, isoenzymes, alternative metabolic pathways, and also from non-enzymatic reactions. In the oxidative branch of the pentose phosphate pathway (oxPPP), 6-phosphogluconolactone hydrolysis into 6-phosphogluconate is catalysed by 6-phosphogluconolactonase (Pgl) but in the absence of the latter, the oxPPP flux is thought to be maintained by spontaneous hydrolysis. However, in Δpgl Escherichia coli, an extracellular pathway can also contribute to pentose phosphate synthesis. This raises question as to whether the intracellular non-enzymatic reaction can compensate for the absence of 6-phosphogluconolactonase and, ultimately, on the role of 6-phosphogluconolactonase in central metabolism. Our results validate that the bypass pathway is active in the absence of Pgl, specifically involving the extracellular spontaneous hydrolysis of gluconolactones to gluconate. Under these conditions, metabolic flux analysis reveals that this bypass pathway accounts for the entire flux into the oxPPP. This alternative metabolic route-partially extracellular-sustains the flux through the oxPPP necessary for cell growth, albeit at a reduced rate in the absence of Pgl. Importantly, these findings imply that intracellular non-enzymatic hydrolysis of 6-phosphogluconolactone does not compensate for the absence of Pgl. This underscores the crucial role of Pgl in ensuring the efficient functioning of the oxPPP.

Formulation of Ascorbic Acid and Betaine-based Therapeutic Deep Eutectic System for Enhanced Transdermal Delivery of Ascorbic Acid.

L-ascorbic acid (AA), a potent antioxidant, is commonly used topically in the pharmaceutical and cosmetic fields. However, the incorporation of AA into topical formulations is difficult because of its highly unstable nature and relatively poor skin permeability. In this study, we propose an alternative strategy for improving the solubility and topical delivery of AA through its conversion to a therapeutic deep eutectic system (THEDES). AA and betaine (Bet)-based THEDESs were prepared at certain molar ratios and characterized using polarized optical microscopy, Fourier transform infrared spectroscopy, and differential scanning calorimetry. Solubility tests showed that AA in the form of THEDES was readily soluble in various polyols (glycerin, 1,3-butylene glycol, dipropylene glycol, and 1,3-propanediol) at a high concentration (approximately 40%). Furthermore, compared to AA alone or the physical mixture of AA and Bet, AA-based THEDES significantly enhanced AA delivery through porcine skin. In an in vivo human study, THEDES-containing serum reduced the markers of aging and induced an even skin tone. These findings indicate the utility of AA and Bet-based THEDES as novel transdermal delivery systems for AA. Furthermore, our approach also showed good extension to developing gluconolactone, a well-known natural antioxidant, and Bet-based THEDES, showing potential application in transdermal delivery systems.

Bio-Based Vitrimeric Silicone Materials with High-Strength, Reprocessable, Healing, and Transparent Properties.

Developing reprocessable polymeric materials from earth-abundant elements and renewable biomass is attractive for dealing with fossil resource crisis and achieving sustainable development. Based on the unique reactivity of biomass-derived gluconolactone, polydimethylsiloxane (PDMS) terminated with glucosamide groups is synthesized and used for preparing a series of silicone boronic ester based vitrimers. The whole preparation process is quite straightforward without any purification required and highly efficient with water as the only byproduct. The mechanical properties of obtained vitrimers can be precisely controlled by adjusting the content of 1,4-benzenediboronic acid or the molecular weight of PDMS precursor, producing boronic ester based vitrimers ranging from soft elastomers to rigid plastics. The obtained vitrimers exhibit excellent thermal stability, robust reprocessability, and efficient healing capacity. By encapsulating green-emitting CsPbBr3 nanocrystals, these materials are fabricated into hydrophobic, transparent, and luminescent coatings, promising for applications in flexible optical devices.

Split-face evaluation: Gluconolactone plus oxybrasion versus gluconolactone plus microneedling. The effects on skin parameters.

BACKGROUND: The application of polyhydroxy acids and alpha-hydroxy acids to the skin is often used in cosmetology. To enhance the effect of gluconolactone chemical peeling, a combined method including water-oxygen oxybrasion or microneedle mesotherapy can be used. OBJECTIVES: To evaluate skin parameters such as hydration, sebum, pH and TEWL after application of a 10% gluconolactone chemical peel in combination with oxybrasion and microneedling. MATERIALS AND METHODS: Twenty-one Caucasian women participated in the study. A series of three split face treatments was carried out at 1-week intervals. Oxybrasion was performed on the right side of the face and micro-needle mesotherapy on the left side. A 10% gluconolactone solution was applied to the entire face. Before the first and third treatments and 2 weeks after the last treatment, skin parameters were evaluated. Photographic documentation was also made before and after the treatment series. RESULTS AND CONCLUSION: Evaluation of skin parameters using Courage & Khazaka 580 Multi Probe Adapter probes (Courage + Khazaka electronic GmbH, Cologne, Germany) showed an increase in hydration and a decrease in sebum, pH and TEWL for both treatments. There were no statistically significant differences between the treatments. Combining chemical peeling of gluconolactone with oxybrasion and microneedle mesotherapy is a good method to seal the hydrolipid barrier and increase skin hydration.

Evaluation of the effects of 10% and 30% gluconolactone chemical peel on sebum, pH, and TEWL.

BACKGROUND: Gluconolactone (GLA) exhibits antioxidant and moisturizing effects. It also presents soothing effects, protects elastin fibers from UV-induced degradation, and improves the function of the skin barrier. AIMS: Evaluation of skin parameters such as pH, transepidermal water loss (TEWL), sebum levels before, during, and after a series of applications of 10% and 30% GLA chemical peel in a split-face model. MATERIALS AND METHODS: The study involved 16 female subjects. Three split-face procedures were performed using two concentrations of GLA solution applied on two sides of the face. The skin parameters were measured before treatments and 7 days after the last procedure at four measurement sites on either side of the face, that is, on the forehead, around the eye, on the cheek, and on the nose wing. RESULTS: Measurement of sebum demonstrated some statistically significant changes between sebum levels in the cheeks after a series of treatments. The pH measurement showed that the pH value was reduced after each treatment at all measurement points. The level of TEWL after treatments was significantly lower around the eyes, on the left forehead, and on the right cheek. There were no significant differences between the use of different concentrations of the GLA solution. CONCLUSIONS: The results of the study show that GLA has a significant influence on lowering skin pH and TEWL. GLA also has seboregulatory properties.

Investigation on the Biological Safety and Activity of a Gluconolactone-Based Lotion for Dermocosmetic Application.

(1) Background: Cosmeceuticals are formulas enriched with active ingredients that exert efficacy on different skin molecular targets. (2) Methods: Cell viability and the absence of potential irritant risk were evaluated on keratinocytes (HaCaT), fibroblasts (NHDF), adipocytes (3T3-L1), sebocytes (PCi-SEB_CAU) and reconstructed human epidermis (RHE), respectively. Several treatments were performed to evaluate the ability of the lotion to stimulate the production of collagen and elastin, stimulate the differentiation of keratinocytes and reduce the number of senescent cells following UVB stimulation. In addition, the modulation of genes involved in the production, storage and accumulation of sebum were investigated. (3) Results: The results obtained demonstrated the biosafety of the formula in all cell lines tested. The 24-h treatment with non-cytotoxic concentrations determined an increase in the expression of the collagen (COL1A1), elastin (ELN) and involucrin (IVL) genes, while a reduction of peroxisome proliferator-activated receptor-gamma (PPARγ) gene expression and a reduction of SA-ßgal-positive cells were found. Moreover, the treatment did not interfere with normal steroid 5-alpha reductase (5RDA3) gene expression levels. (4) Conclusions: Data collected demonstrated the biosafety of the lotion, the non-comedogenic property and a multi targets anti-aging effect. In particular, data collected on the booster lotion make it a valid way to counteract the pore dilatation aging related.

Gluconolactone Alleviates Myocardial Ischemia/Reperfusion Injury and Arrhythmias via Activating PKCε/Extracellular Signal-Regulated Kinase Signaling.

Gluconolactone (D-glucono-1,5-lactone or GDL) is a food additive which presents in dietary products such as tofu, yogurt, cheese, bread, wine, etc. GDL has long been considered as a free radical scavenger; however, its role in cardioprotection remains elusive. In this study, using a mouse model of myocardial ischemia/reperfusion (I/R) injury and a model of hypoxia/reoxygenation (H/R) in neonatal rat cardiomyocytes (NRCM), we explored the role of GDL in I/R injury. We found that GDL (5 mg/kg, i.p.) attenuated myocardial I/R injury as evidenced by decreased infarct size, release of cardiac injury markers and apoptosis. Additionally, GDL decreased reperfusion-induced arrhythmias and oxidative stress. These effects were also observed in parallel in vitro studies. Mechanistically, we found that GDL treatment was strongly associated with activation of pro-survival extracellular signal-regulated kinase (ERK) signaling both in vivo and in vitro, and pharmacological inhibition of ERK signaling via U0126 attenuated GDL-induced cardioprotection against H/R injury in NRCM cells. To reveal how GDL regulates ERK signaling, we predicted the putative targets of GDL by Swiss Target Prediction, and protein kinase C (PKC) emerged as the most promising target for GDL. By pharmacological intervention and immunofluorescence, we found that PKCε, an important member of the PKC family, was activated after GDL treatment in heart, thereby leading to ERK activation and cardioprotection against I/R injury. Taken together, our results demonstrated that GDL acts as a potent activator of PKCε and, thus, provides cardioprotection against I/R injury via activation of ERK signaling.

Dissolution of Zinc Oxide Nanoparticles in the Presence of Slow Acid Generators.

We describe a preliminary investigation of the dissolution dynamics of zinc oxide nanoparticles in the presence of cyclic esters (δ-gluconolactone and propanesultone) as slow acid generators. The particles dissolution is monitored by means of turbidimetry and correlated with the evolution of pH over time. The results could be of interest for the design of chemically programmable colloidal systems.

Corneometric evaluation of skin moisture after application of 10% and 30% gluconolactone.

BACKGROUND: Dry skin, caused by improper care or genetic conditions, can affect people of all ages. Skin hydration is determined its lipid content, which inhibits water loss from the epidermis, as well as other substances such as polyhydroxy acids and gluconolactone that can bind water. The aim of this study was to evaluate skin hydration after the application of 10% and 30% gluconolactone solution in a split face model. MATERIALS AND METHODS: Sixteen healthy women were qualified for the study. Three split face treatments were performed, with 10% and 30% gluconolactone solution applied to two sides of the face. Skin moisture was measured before each treatment and a week after the last treatment at three measurement sites on either side of the face, that is, on the forehead, around the eye and on the cheek. RESULTS: Corneometric measurements showed a significant increase in facial skin hydration after gluconolactone treatment. No significant differences were observed between the application of 10% and 30% solution. CONCLUSION: Gluconolactone is a moisturizing substance which works well in dry skin care.

Cultivation of human skin cells under physiological oxygen concentration modulates expression of skin significant genes and response to hydroxy acids.

Physiological oxygen concentration (physioxia) ranges from 1 to 8% in human tissues while many researchers cultivate mammalian cells under an atmospheric concentration of 21% (hyperoxia). Oxygen is one of the significant gases which functions in human cells including energy production in mitochondria, metabolism in peroxidase, and transcription of various genes in company with HIF (Hypoxia-inducible factors) in the nucleus. Thus, mammalian cell culture should be deliberated on the oxygen concentration to mimic in vivo physiology. Here, we studied if the cultivation of human skin cells under physiological conditions could affect skin significant genes in barrier functions and dermal matrix formation. We further examined that some representative active ingredients in dermatology such as glycolic acid, gluconolactone, and salicylic acid work in different ways depending on the oxygen concentration. Taken together, we present the importance of oxygen concentration in skin cell culture for proper screening of novel ingredients as well as the mechanistic study of skin cell regulation.

Glycolipid-Based Oleogels and Organogels: Promising Nanostructured Structuring Agents.

Over the past few decades, the scientific community is actively involved in the development of edible structuring agents suitable for food, cosmetics, agricultural, pharmaceutical, and biotechnology applications. In particular, edible oil structuring using simple amphiphiles would be the best alternative for the currently used trans and saturated fatty acids, which cause deleterious health effects and cardiovascular problems. In this report, we have made an attempt to address the aforementioned consequences, by synthesizing a new class of structuring agents by a judicious combination of δ-gluconolactone and ricinoleic acid, compounds classified as GRAS, using simple steps in good yield. To our delight, the synthesized glycolipids self-assemble in a wide variety of vegetable oils and commercially viable glycerol, ethylene glycol, and polyethylene glycol via various intermolecular interactions to form a gel. The morphology of molecular gels was investigated by optical microscopy and FESEM analysis, which reveal the existence of a tubular architecture with a diameter ranging from 75 to 150 nm. Rheological studies disclosed the viscoelastic nature, thermal processability, and thixotropic behavior of both oleogels and organogels. Altogether, self-assembled oleogel and organogel reported in this paper would potentially be used in food, agricultural, cosmetics, pharmaceutical, and biotechnological applications.

Enzyme kinetics and inhibition parameters of human leukocyte glucosylceramidase.

Glucosylceramidase (GCase) is a lysosomal enzyme that catalyzes the cleavage of ß-glucosidic linkage of glucocerebroside (GC) into glucose and ceramide; thereby, plays an essential function in the degradation of complex lipids and the turnover of cellular membranes. The growing list of 460 mutations in the gene coding for it-glucosylceramidase beta acid 1 (GBA1)-is reported to abolish its catalytic activity and decrease its enzyme stability, associating it with severe health conditions such as Gaucher disease (GD), Parkinson Disease (PD) and Dementia with Lewy bodies (DLB). Although the three-dimensional structure of wild type glucosylceramidase is elucidated, little is known about its features in human cells. Moreover, alternative sources of GCase that prove to be effective in the treatment of diseases with enzyme treatment therapies, impose the need for a simple and cost-effective procedure to study the enzyme behavior. This work, for the first time, shows a well-established, yet simple, cost- and time-efficient protocol for the study of GCase enzyme in human leukocytes by the artificial substrate p-Nitrophenyl-ß-D-glucopyranoside (PNPG). Characterization of the enzyme in human leukocytes for activation parameters (optimal pH, Km, and Vmax) and enzyme inhibition was done. The results indicate that the optimum pH of GCase enzyme with PNPG is 5.0. The Km and Vmax values are 12.6mM and 333 U/mg, respectively. Gluconolactone competitively inhibits GCase, with a Ki value of 0.023 mM and IC50 of 0.047 mM. Glucose inhibition is uncompetitive with a Ki of 1.94 mM and IC50 of 55.3 mM. This is the first report for the inhibitory effect of glucose, δ-gluconolactone on human leukocyte GCase activity.

Evaluation of the Neurobehavioural Toxic Effects of Taurine, Glucuronolactone, and Gluconolactone Used in Energy Drinks in Young Rats.

OBJECTIVES: The neurotoxic effects of food additives used in energy drinks have been investigated since the 1900s but safety concerns are rising and reassurance via safety testing in animals is demanded by the public. Rigorous safety testing is performed for dose optimisation and duration of treatment and to detect the methods to assess changes in mood and behaviour. Hence, we studied the neurobehavioral effects of selected food additives used in energy drinks and their combination in rats when consumed in high doses. MATERIALS AND METHODS: Young Sprague Dawley rats were divided into six groups. Group 1 was treated with the vehicle, group 2 was treated with 25 mg/kg p.o. caffeine, group 3 was treated with 5 mg/kg p.o. glucuronolactone, group 4 was treated with 8 mg/kg p.o. taurine, group 5 was treated with 84 mg/kg p.o. gluconolactone, and group 6 was treated with a combination of the three food additives. Neurobehavioral changes were evaluated on days 7, 14, and 21 using behavioural parameters. Neurobehavioral scoring and neurotransmitter estimation in rat brain tissue was performed on day 21. RESULTS: Significant changes were observed in the neurobehavioral parameters and neurobehavioural scoring in group 4 and group 6, compared with the control group (p<0.001). Furthermore, the significant decreases in neurotransmitter levels in the brains of rats that were treated with food additives indicated the neurotoxic effects of these substances. CONCLUSION: This study elaborated the neurobehavioral effects of selected food additives, namely glucuronolactone, taurine, and gluconolactone, when administered orally for 21 days in young rats. The highest toxic effects, including alterations in neurotransmitter levels, were observed in animals treated with a combination of food additives at high doses.

Feasibility of Using Gluconolactone, Trehalose and Hydroxy-Propyl Gamma Cyclodextrin to Enhance Bendroflumethiazide Dissolution Using Lyophilisation and Physical Mixing Techniques.

PURPOSE: Hydrophobic drugs are facing a major challenge in dissolution rate enhancement and solubility in aqueous solutions; therefore, a variety of methods have been used to improve dissolution rate and/or solubility of bendroflumethiazide as a model hydrophobic drug. METHODS: In this study, two main methods (physical mixing and lyophilisation) were used with gluconolactone, hydroxyl propyl γ-ccyclodextrin, and trehalose to explore this challenge. Bendroflumethiazide, practically insoluble in water, was mixed with one of the three excipients gluconolactone, hydroxyl propyl γ-cyclodextrin, and trehalose in three different ratios 1:1, 1:2, 1:5. To the best of our knowledge, the dissolution of the drug has not been previously enhanced by using either these methods or any of the used excipients. Samples containing drug and each of the excipients were characterized via dissolution testing, Fourier Transform infra-red spectroscopy, differential scanning calorimetry, and scanning electron microscopy. RESULTS: The used methods showed a significant enhancement in dug dissolution rate; physical mixing significantly, p < 0.05, increased the percentage of the drug released with time; for example, bendroflumethiazide dissolution in distilled water was improved from less than 20% to 99.79% within 90 min for physically mixed drug-cyclodextrin 1:5. The lyophilisation process was enhanced and the drug dissolution rate and the highest drug dissolution was achieved for (drug-gluconolactone 1:1) with 98.98% drug release within 90 min. CONCLUSIONS: the physical mixing and freeze drying processes significantly increased the percentage of drug release with time.

Industrial robustness linked to the gluconolactonase from Zymomonas mobilis.

The physiological characteristics and the potential gluconolactone production of the gluconolactonase-deficient strain, Zymomonas mobilis ZM4 gnlΔ, were investigated via growth inhibitory assay and biotransformation of glucose and fructose into gluconolactone and sorbitol, respectively. The results of ethanol fermentation studies performed in the presence of high concentration of glucose (>200 g l-1) under fermentative or aerobic conditions indicated that a significant reduction of volumetric ethanol productivity from the strain of ZM4 gnlΔ was noticeable due to the reduced rates of specific growth, sugar uptake, and biomass yield as compared with those of the parental strain ZM4. The biotransformation prepared at pH 6.0 using the permeabilized cell indicated that gluconic acid from ZM4 gnlΔ was still produced as a major product (67 g l-1) together with sorbitol (65 g l-1) rather than gluconolactone after 24 h. Only small amount of gluconolactone was transiently overproduced up to 9 g l-1, but at the end of biotransformation, all gluconolactone were oxidized into gluconic acid. This indicated that autolysis of gluconolactone at the pH led to such results despite under gluconolactonase inactivation conditions. The physiological characteristics of ZM4 gnlΔ was further investigated under various stress conditions, including suboptimal pH (3.5~6.0), temperature (25~40 °C), and presence of growth inhibitory molecules including hydrogen peroxide, ethanol, acetic acid, furfural, and so forth. The results indicated that ZM4 gnlΔ was more susceptible at high glucose concentration, low pH of 3.5, and high temperature of 40 °C and in the presence of 4 mM H2O2 comparing with ZM4. Therefore, the results were evident that gluconolactonase in Z. mobilis contributed to industrial robustness and anti-stress regulation.

Cr(VI) reduction by gluconolactone and hydrogen peroxide, the reaction products of fungal glucose oxidase: Cooperative interaction with organic acids in the biotransformation of Cr(VI).

The Cr(VI) reducing capability of growing cells of the environmental A. tubingensis Ed8 strain is remarkably efficient compared to reference strains A. niger FGSC322 and A. tubingensis NRRL593. Extracellular glucose oxidase (GOX) activity levels were clearly higher in colonies developed in solid medium and in concentrated extracts of the spent medium of liquid cultures of the Ed8 strain in comparison with the reference strains. In addition, concentrated extracts of the spent medium of A. tubingensis Ed8, but not those of the reference strains, exhibited the ability to reduce Cr(VI). In line with this observation, it was found that A. niger purified GOX is capable of mediating the conversion of Cr(VI) to Cr(III) in a reaction dependent on the presence of glucose that is stimulated by organic acids. Furthermore, it was found that a decrease in Cr(VI) may occur in the absence of the GOX enzyme, as long as the reaction products gluconolactone and hydrogen peroxide are present; this conversion of Cr(VI) is stimulated by organic acids in a reaction that generates hydroxyl radicals, which may involve the formation of an intermediate peroxichromate(V) complex. These findings indicated that fungal glucose oxidase acts an indirect chromate reductase through the formation of Cr(VI) reducing molecules, which interact cooperatively with other fungal metabolites in the biotransformation of Cr(VI).

The dissolution enhancement of piroxicam in its physical mixtures and solid dispersion formulations using gluconolactone and glucosamine hydrochloride as potential carriers.

The solid dispersion technique is one of the most effective methods for improving the dissolution rate of poorly water-soluble drugs; however this is reliant on a suitable carrier and solvent being selected. The work presented explores amino sugars (d-glucosamine HCl and d-gluconolactone) as potential hydrophilic carriers to improve dissolution rate of a poorly water-soluble drug, piroxicam, from physical mixtures and solid dispersion formulations. Solid dispersions of the drug and carrier were prepared using different ratios by the conventional solvent evaporation method. Acetone was used as solvent in the preparation of solid dispersions. Physical mixtures of piroxicam and carrier were also prepared for comparison. The properties of all solid dispersions and physical mixtures were studied using a dissolution tester, Fourier transform infrared, XRD, SEM and differential scanning calorimetry. These results showed that the presence of glucosamine or gluconolactone can increase dissolution rate of piroxicam compared to pure piroxicam. Glucosamine or Gluconolactone could be used as carrier in solid dispersion formulations and physical mixtures to enhance the dissolution rate. Solid state studies showed that no significant changes occurred for piroxicam in physical mixtures and solid dispersion.

Novel poly(ethylene oxide)-b-poly(propylene oxide) copolymer-glucose conjugate by the microwave-assisted ring opening of a sugar lactone.

In this work, we investigated for the first time the conjugation of gluconolactone to a poly(ethylene oxide)-poly(propylene oxide) block copolymer by a microwave-assisted ring opening reaction. The glucosylated copolymer was obtained with high yield (90%). A conjugation extent of approximately 100% was achieved within 15 min. The modification reduced the critical micellar concentration and increased the size of the micelles. The agglutination of the modified polymeric micelles by a soluble lectin that binds glucose confirmed the recognizability of the modified nanocarrier. Finally, the solubilization of darunavir, an anti-HIV protease inhibitor, showed a sharp increase of the aqueous solubility from 91 microgram/mL to 14.2 and 18.9 mg/mL for 10% w/v pristine and glucosylated polymeric micelles, respectively.

Assessment of the enzymatic hydrolysis profile of cellulosic substrates based on reducing sugar release.

The activity profile of a 1:0.30 mixture of Celluclast 1.5L FG and Novozym 188 (Novozymes) was investigated using Whatman #1 filter paper (W1FP) as a single substrate for hydrolysis. The procedure was based on the ability of the enzymes to release total (RS(Tot)), insoluble (RS(Insol)) and soluble (RS(Sol)) reducing sugars from W1FP. RS(Insol) was used to estimate endoglucanase (EnG) activity whereas exoglucanases (ExG) were assessed by measuring RSSol in the presence of δ-gluconolactone. Finally, the ß-glucosidase (ßG) activity was derived from the difference between RS(Sol) measurements in the presence and absence of δ-gluconolactone. When this analytical procedure was applied to W1FP using 9.64 mg mL(-1) of the enzyme mixture, the relative contributions of EnG, ExG and ßG to the total cellulase activity were 63.28%, 12.02% and 24.70%, respectively. Also, this ratio changed with changes in the enzyme loading, giving a new insight into the synergy that exists among the enzymes.