Preclinical studies on melanogenesis proteins using a resveratrol-nanoformula as a skin whitener.

A naturally occurring polyphenol called trans-resveratrol has received a lot of attention due to its possible health advantages for humans. The low solubility of trans-resveratrol and its isomerization upon UV exposure strongly limit its application as a skin-whitening agent. In the present study, to increase trans-resveratrol solubility, a new nanoformula was created by combining hydrophilic surfactants and oils. Trans-Resveratrol nanoformula has been prepared, characterized, and applied as a skin-whitening agent on the dorsal skin of Guinea pigs. The optimized trans-resveratrol nanoformula with a particle size of 63.49 nm displayed a single peak and a polydispersity index [0.36 ± 0.02]. In addition, the zeta potential of the optimized formula was -30.4 mV, confirming the high stability of this nanoformula. The melanin contents in the trans-resveratrol nanoformula-treated group were substantially lower than those of the control and the blank nanoformula-treated groups after staining of the dorsal skins [black areas] of guinea pigs with Fontana Mountain dye. The pigmentation index in the control, blank nanoformula, and optimized trans-resveratrol nanoformula were 329.4 ± 36.9, 335.8 ± 71.4, and 124.8 ± 19.6 respectively. Confirming this finding, immunohistochemistry analysis of skin tissues revealed that the expressions of melanogenesis-regulating proteins such as tyrosinase and microphthalmia-associated transcription factor were down-regulated. The safety of topical application of trans-resveratrol nanoformula was validated by no changes in free radical levels and oxidative stress markers proteins in the livers and kidneys of guinea pigs at the end of the experiment. Conclusions: A novel trans-resveratrol nanoformula as well as the mechanism whereby it promotes skin whitening effects were presented. Furthermore, the study illustrated that trans-resveratrol nanoformula is safe, non-toxic, and can be applied for skin whitening, although more research on human skin is needed.

Evaluating the Antioxidants, Whitening and Antiaging Properties of Rice Protein Hydrolysates.

Plant-derived protein hydrolysates have potential applications in nutrition. Rice protein hydrolysates (RPHs), an excellent source of proteins, have attracted attention for the development of cosmeceuticals. However, few studies have reported the potential application of RPH in analysis, and this study examined their antioxidant activities and the inhibitory activities of skin aging enzymes. The results indicated that the total phenolic and flavonoid concentrations were 2.06 ± 0.13 mg gallic acid equivalent/g RPHs and 25.96 ± 0.52 µg quercetin equivalent/g RPHs, respectively. RPHs demonstrated dose-dependent activity for scavenging free radicals from 1,1-diphenyl-2-picrylhydrazyl [half-maximal inhibitory concentration (IC50) = 42.58 ± 2.1 mg/g RPHs] and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (IC50 = 2.11 ± 0.88 mg/g RPHs), dose-dependent reduction capacity (6.95 ± 1.40 mg vitamin C equivalent/g RPHs) and oxygen radical absorbance capacity (473 µmol Trolox equivalent/g RPHs). The concentrations of the RPH solution required to achieve 50% inhibition of hyaluronidase and tyrosinase activities were determined to be 8.91 and 107.6 mg/mL, respectively. This study demonstrated that RPHs have antioxidant, antihyaluronidase, and antityrosinase activities for future cosmetic applications.

Immobilization of peroxidase on textile carrier materials and their application in the bleaching of colored whey.

Textiles represent promising support materials for enzymes. The goal of the present work was to investigate the immobilization of commercial peroxidase on a polyester needle felt and the repeated use in the gentle degradation of norbixin in whey from dairy cheese as a practical application. High enzyme loads were obtained by a 2-step immobilization procedure. First, the number of functional groups on the textile surface was increased by a modification with amino-functional polyvinylamine. Second, the enzyme was immobilized by using 2 types of crosslinking agents. Due to the iron content of peroxidase, inductively coupled plasma-optical emission spectrometry was used for the quantitative determination of the enzyme load on the textile. The enzyme activity was evaluated using common 2,2'-azino-di-(3-ethylbenzthiazoline-6-sulfonic acid) assay for peroxidases. By the variation of enzyme input and crosslinker concentration, a maximal enzyme load of 80 mg/g of textile was achieved, and a maximum specific activity of 57 U/g of textile. For the visualization of the enzyme on the fiber surface, fluorescence microscopy as well as scanning probe microscopy were used. The immobilized peroxidase showed significant activity, even after 50 reuse cycles. In addition, the potential of the new support and enzyme combination in commercial whey bleaching was demonstrated successfully on a 10-L scale.

A DyP-Type Peroxidase of Pleurotus sapidus with Alkene Cleaving Activity.

Alkene cleavage is a possibility to generate aldehydes with olfactory properties for the fragrance and flavor industry. A dye-decolorizing peroxidase (DyP) of the basidiomycete Pleurotus sapidus (PsaPOX) cleaved the aryl alkene trans-anethole. The PsaPOX was semi-purified from the mycelium via FPLC, and the corresponding gene was identified. The amino acid sequence as well as the predicted tertiary structure showed typical characteristics of DyPs as well as a non-canonical Mn2+-oxidation site on its surface. The gene was expressed in Komagataella pfaffii GS115 yielding activities up to 142 U/L using 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) as substrate. PsaPOX exhibited optima at pH 3.5 and 40 °C and showed highest peroxidase activity in the presence of 100 µM H2O2 and 25 mM Mn2+. PsaPOX lacked the typical activity of DyPs towards anthraquinone dyes, but oxidized Mn2+ to Mn3+. In addition, bleaching of ß-carotene and annatto was observed. Biotransformation experiments verified the alkene cleavage activity towards the aryl alkenes (E)-methyl isoeugenol, α-methylstyrene, and trans-anethole, which was increased almost twofold in the presence of Mn2+. The resultant aldehydes are olfactants used in the fragrance and flavor industry. PsaPOX is the first described DyP with alkene cleavage activity towards aryl alkenes and showed potential as biocatalyst for flavor production.

Biobleaching of mechanical paper pulp using Streptomyces rutgersensis UTMC 2445 isolated from a lignocellulose-rich soil.

AIMS: The utilization of micro-organisms in pulp and paper industries has proved biobleaching technology as an environmentally friendly alternative to the conventional approach. In this paper, the effect of actinobacterial fermentation broth on pulp biobleaching has been investigated. METHODS AND RESULTS: Actinobacterial colonies were isolated from lignocellulose-rich soil samples and screened for xylanase production and bleaching activity. The most efficient isolate in bleaching activity showed 100% similarity to Streptomyces rutgersensis. Pulp treatment with 5-day fermentation broth of this strain showed up to 7% increase in brightness (30°C for 6 h, pH (5-7)) compared to untreated (control) pulp. Also, after 60 min biotreatment, significant reduction (12·5%) in consumption of bleaching chemicals was achieved to obtain final brightness of 55%. CONCLUSION: Actinobacterial fermentation broth can be considered as a rich source of effective biobleaching agents which may be considered as environmental friendly and cost-effective technique in comparison with traditional method. SIGNIFICANCE AND IMPACT OF THE STUDY: Our findings showed ability of S. rutgersensis UTMC 2445 in bleaching chemomechanical paper pulp. Also, two strains of Saccharothrix, a rare actinobacterium, with biobleaching activity were introduced. In the proposed method, there is no need to use purified enzymes, and biobleaching process can be done using the fermentation broth.

Xylanases from marine microorganisms: A brief overview on scope, sources, features and potential applications.

Global economic growth often leads to depletion of raw materials and generation of greenhouse gases, as industry manufactures goods at ever increasing levels to keep up with the demand. The currently implemented production processes mostly rely on non-renewable resources, they suffer from high energy consumption, and generate waste that often has a negative environmental impact. Eco-friendly production methods are therefore intensely searched for. Among them, enzyme-based processes are appealing, because of their high substrate and reaction specificity and the relatively mild operation conditions required by these catalysts. In addition, renewable raw materials that allow sustainable production processes are also widely explored. Marine xylanases, which catalyze the hydrolysis of xylan, the major component of lignocellulose, are promising biocatalysts. Since they are produced by microorganisms that thrive in a wide variety of environmental conditions, the enzymes may be active at widely different ranges of pH, temperature, and salt concentrations. These properties are important for their successful application in various industrial processes, such as production of bioethanol, bleaching of paper and pulp, and in the food and feed sector. The present work gives a brief overview of marine sources of xylanases, their classification and features, and of the potential applications of these marine enzymes, especially in sustainable processes in the scope of circular economy.

Influence of a short-time antioxidant application on the dentin bond strength after intracoronal bleaching.

This study evaluated the effects of an antioxidant application on the compromised bond strength of an adhesive to dentin bleached with 35% hydrogen peroxide. The dentin surfaces of the pulp chambers of 70 human third molars were ground, and the specimens were assigned randomly into seven groups, as follows: (a) control (unbleached); (b) bleached for 45 min, bonded immediately; (c) bleached for 45 min, treated with sodium ascorbate (SA) for 2 min, and bonded; (d) bleached for 45 min, bonded after 2 weeks; (e) bleached for 12 days, bonded immediately; (f) bleached for 12 days, treated with SA for 10 min, and bonded; and (g) bleached for 12 days, bonded after 2 weeks. In each group, the multimode adhesive was applied in etch-and-rinse and self-etch modes. The dentin surfaces were covered with a resin-based composite, and the bonded specimens were sectioned to produce composite-dentin sticks. The sticks were attached to a testing machine and subjected to a tensile force, and the representative specimens were examined via scanning electron microscopy. The bond strength was not affected by the application period of the bleaching agent. Both bleaching treatments significantly reduced the bond strength to the dentin in the self-etch or etch-and-rinse mode when compared with the control group. The bond strengths returned to normal levels with the SA applications or by waiting 2 weeks, regardless of the application period of the bleaching gel. The adhesive revealed a higher bond strength in the etch-and-rinse mode than in the self-etch mode.

The presence of fecal test soil protects bacteria during cleaning/disinfection.

Reusable medical devices (RMDs) must be reprocessed between uses to render them safe for each use and each patient. Cleaning used devices removes organic and inorganic soil making them either safe for reuse or ready for disinfection/sterilization depending on the device. Although cleaning is an important step in a RMD's life cycle, it is not always a priority during device design. In addition, when performing cleaning validation, it is recommended that the manufacturer takes into consideration, what the most appropriate or worst case conditions are in terms of type of soil or the presence of bacteria. This study compared the ability of three different cleaning/disinfecting agents (water, alcohol, and bleach) to remove bacteria and fecal test soil from two different polymers: polypropylene and ultrahigh molecular weight polyethylene (UHMWPE) with two different roughness. There were some differences in the effects of the cleaning/disinfecting agents, the materials, and the roughness depending on the particular circumstances. However, the most consistent effect on the removal of bacteria was the presence of soil, which protected the bacteria from being removed. Conversely, the presence of bacteria played little role in the removal of soil. Although the interactions between material type and roughness, soil type, and bacteria are complicated, they should be taken into account during device design and reprocessing validation to create a device that is easy and safe to use. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 1706-1710, 2019.

Optimization of physical and morphological regime for improved cellulase free xylanase production by fed batch fermentation using Aspergillus niger (KP874102.1) and its application in bio-bleaching.

The physiological and morphological changes were extensively studied during fed batch fermentation using newly isolated Aspergillus niger (KP874102.1). Significantly higher xylanase production was possible through optimization of environmental stresses by fed batch process. The fed batch fermentation was carried out for improved xylanase production (2524 U) where initial xylan was kept 1.5 g/L in the production medium. However, 3 g/L of xylan with 50 mM K2HPO4 having pH-7 was consecutively fed at 72 and 120 h of fermentation. K2HPO4 showed significant role both the morphology of the microorganism and produces enzymes in fed batch fermentation. During feeding phase, the pH was found in the range of 6.5 to 7 which was used as marker for the fed batch process. The crude enzyme was used for the bio-bleaching of banana pulp.

High level extracellular production of a recombinant alkaline catalase in E. coli BL21 under ethanol stress and its application in hydrogen peroxide removal after cotton fabrics bleaching.

The effects of induction parameters, osmolytes and ethanol stress on the productivity of the recombinant alkaline catalase (KatA) in Escherichia coli BL21 (pET26b-KatA) were investigated. The yield of soluble KatA was significantly enhanced by 2% ethanol stress. And a certain amount of Triton X-100 supplementation could markedly improved extracellular ratio of KatA. A total soluble catalase activity of 78,762U/mL with the extracellular ratio of 92.5% was achieved by fed-batch fermentation in a 10L fermentor, which was the highest yield so far. The purified KatA showed high stability at 50°C and pH 6-10. Application of KatA for elimination of H2O2 after cotton fabrics bleaching led to less consumption of water, steam and electric power by 25%, 12% and 16.7% respectively without productivity and quality losing of cotton fabrics. Thus, the recombinant KatA is a promising candidate for industrial production and applications.

Alkaliphilic endoxylanase from lignocellulolytic microbial consortium metagenome for biobleaching of eucalyptus pulp.

Enzymatic pre-bleaching by modification of pulp fibers with xylanases is an attractive approach to reduce the consumption of toxic bleaching chemicals in the paper industry. In this study, an alkaliphilic endoxylanase gene was isolated from metagenomic DNA of a structurally stable thermophilic lignocellulose-degrading microbial consortium using amplification with conserved glycosyl hydrolase family 10 primers and subsequent genome walking. The full-length xylanase showed 78% sequence identity to an endo-beta-1,4-xylanase of Clostridium phytofermentans and was expressed in a mature form with an N-terminal His6 tag fusion in Escherichia coli. The recombinant xylanase Xyn3F was thermotolerant and alkaliphilic, working optimally at 65-70 degrees C with an optimal pH at 9- 10 and retaining >80% activity at pH 9, 60 degrees C for 1 h. Xyn3F showed a Vmax of 2,327 IU/mg and Km of 3.5 mg/ml on birchwood xylan. Pre-bleaching of industrial eucalyptus pulp with no prior pH adjustment (pH 9) using Xyn3F at 50 IU/g dried pulp led to 4.5-5.1% increase in final pulp brightness and 90.4-102.4% increase in whiteness after a single-step hypochlorite bleaching over the untreated pulp, which allowed at least 20% decrease in hypochlorite consumption to achieve the same final bleaching indices. The alkaliphilic xylanase is promising for application in an environmentally friendly bleaching step of kraft and soda pulps with no requirement for pH adjustment, leading to improved economic feasibility of the process.

One-copper laccase-related enzyme from Marasmius sp.: purification, characterization and bleaching of textile dyes.

In the culture filtrate of a Marasmius sp. strain isolated in Indonesia during a screening for fungi with the ability to decolorize textile dyes, two laccase-related enzymes (laccase-related enzyme I and II) were detected. Laccase-related enzyme I was purified to homogeneity by ion exchange and hydrophobic interaction chromatography. The native enzyme was shown to have a molecular mass of 53 kDa, an N-terminal amino acid sequence characteristically seen in laccases and an isoelectric point of pH 3.8. The enzyme accepts typical laccase substrates including 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), syringaldazine and guaiacol, but has no tyrosinase activity. The pH optimum is at pH 3.0 for ABTS and at 6.0 for syringaldazine and the enzyme is stable up to pH 10. The UV/vis spectrum of the laccase-related enzyme is non-typical for laccases and metal content analysis revealed that the enzyme contains only a single copper atom per enzyme molecule. This suggests that this enzyme could be related to the group of the so-called "white" laccases, however, no zinc or any other metal ion could be detected in this enzyme, suggesting that the enzyme is a unique laccase-related enzyme. Comparison of the bleaching activity of the whole fungus with that of the isolated laccase-related enzyme showed that this enzyme is the major bleaching enzyme produced by this Marasmius sp. strain and was able to bleach violet, red, orange and yellow dyes in addition to a number of blue dyes.

Treatment of bleached wool with trans-glutaminases to enhance tensile strength, whiteness, and alkali resistance.

Trans-glutaminases is known as a cross-linking enzyme for proteins. Wool is a proteinous fiber conventionally is treated through several processes to obtain the desirable characteristics. Bleaching is also one of the most important processes usually carried out by using an oxidizing agent in a conventional method. The tensile strength of wool yarns was reduced as a consequence of oxidative bleaching. Here, with the help of microbial trans-glutaminases (m-TGases), a novel bleaching process was disclosed in a way to obtain a bleached wool yarn with no significant reduction in the tensile strength. The results confirmed that the bleached wool yarns with H(2)O(2) could be modified by m-TGases post-treatment. The m-TGases treatment on the bleached wool yarns improved the tensile strength and whiteness along with the higher alkali resistance.

Optimization of xylanase production from Aspergillus niger for biobleaching of eucalyptus pulp.

A crude endo-xylanase produced by Aspergillus niger BCC14405 was investigated for its potential in pre-bleaching of chemical pulp from eucalyptus. The optimal fermentation conditions on the basis of optimization using response surface methodology included cultivation in a complex medium comprising wheat bran, rice bran, and soybean meal supplemented with yeast extract, glucose, peptone, and lactose with a starting pH of 6.0 for 7 d. This resulted in production of 89.5 IU/mL of xylanase with minor cellulase activity. Proteomic analysis using LC/MS/MS revealed that the crude enzyme was a composite of hemicellulolytic enzymes, including endo-ß-1,4-xylanase and other hemicellulolytic enzymes attacking arabinoxylan and mannan. Pretreatment of the pulp at a xylanase dosage of 10 IU/g increased the brightness ceiling after the C-Eop-H bleaching step up to 3.0% using a chlorine charge with a C-factor of 0.16-0.20. Xylanase treatment also led to reduction in chlorine charge of at least 20%, with an acceptable brightness level. The enzyme pretreatment resulted in a slight increase in pulp viscosity, suggesting an increase in relative cellulose content. The crude enzyme was potent in the enzyme-aided bleaching of chemical pulp in an environmentally friendly pulping process.

Bleach-boosting effect of crude xylanase from Bacillus stearothermophilus SDX on wheat straw pulp.

Pretreatment of wheat straw pulp using cellulase-free xylanase produced from Bacillus stearothermophilus SDX at 60°C for 120min resulted in 4.75% and 22.31% increase in brightness and whiteness, respectively. Enzyme dose of 10U/g of oven dried pulp at pH 9 decreased the kappa number and permanganate number by 7.14% and 5.31%, respectively. Further chlorine dioxide and alkaline bleaching sequences (CDED(1)D(2)) resulted in 1.76% and 3.63% increase in brightness and whiteness, respectively. Enzymatic prebleaching of pulp decreased 20% of chlorine consumption without any decrease in brightness. Improvement in various pulp properties like viscosity, burst factor, burstness, breaking length, double fold, gurley porosity, tear factor, and tearness were also observed after bleaching of xylanase treated wheat straw pulp.

Bio-conventional bleaching of wheat straw soda-AQ pulp with crude xylanases from SH-1 NTCC-1163 and SH-2 NTCC-1164 strains of Coprinellus disseminatus to mitigate AOX generation.

Two novel cellulase-poor xylanases from Coprinellus disseminatus SH-1 NTCC-1163 (enzyme-A) and SH-2 NTCC-1164 (enzyme-B) produced under solid-state fermentation mitigated kappa number of wheat straw soda-AQ pulps by 24.38 and 27.94% respectively after XE stages. The release of reducing sugars and chromophores was highest for both the enzymes at 10IU/g and reaction time 180min for 55°C at variable consistencies that is, 10% for enzyme-A and 5% for enzyme-B. (A)XECEHH and (B)XECEHH sequences improved brightness by 5.17 and 2.58% respectively at 4.5% chlorine charge. AOX in (A)XECEHH and (B)XECEHH sequences reduced by 56.11 and 55.75% respectively at 4.5% chlorine charge and 68.34 and 67.98% respectively at 2.25% chlorine charge respectively compared to control. Both the enzymes showed improvement in double fold and tear index with a decrease in burst and tensile index. SEM showed peeling, cracking and delamination in fibers due to enzyme treatment thus facilitating the penetration of bleach chemicals.

Characterization of a family GH5 xylanase with activity on neutral oligosaccharides and evaluation as a pulp bleaching aid.

A new bacterial xylanase belonging to family 5 of glycosyl hydrolases was identified and characterized. The xylanase, Xyn5B from Bacillus sp. strain BP-7, was active on neutral, nonsubstituted xylooligosaccharides, showing a clear difference from other GH5 xylanases characterized to date that show a requirement for methyl-glucuronic acid side chains for catalysis. The enzyme was evaluated on Eucalyptus kraft pulp, showing its effectiveness as a bleaching aid.