The in vitro effects of artificial and natural sweeteners on the immune system using whole blood culture assays

This article investigates the effects of commercially available artificial (aspartame, saccharin, sucralose) and natural sweeteners (brown sugar, white sugar, molasses) on the immune system. Human whole blood cultures were incubated with various sweeteners and stimulated in vitro with either phytohemagglutinin or endotoxin. Harvested supernatants were screened for cytotoxicity and cytokine release. Results showed that none of the artificial or natural sweeteners proved to be cytotoxic, indicating that no cell death was induced in vitro. The natural sweetener, sugar cane molasses (10 ug=mL), enhanced levels of the inflammatory biomarker IL-6 while all artificial sweeteners (10 ug=mL) revealed a suppressive effect on IL-6 secretion (P<0.001). Exposure of blood cells to sucralose-containing sweeteners under stimulatory conditions reduced levels of the biomarker of humoral immunity, Interleukin-10 (P<0.001). The cumulative suppression of Interleukin-6 and Interleukin-10 levels induced by sucralose may contribute to the inability in mounting an effective humoral response when posed with an exogenous threat.Web of Scienc

Biotechnological production and application of fructooligosaccharides

Currently, prebiotics are all carbohydrates of relatively short chain length. An important group is the fructooligosaccharides, which are a special kind of prebiotics associated to their selective stimulation of the activity of certain groups of colonic bacteria that have a positive and beneficial effect on intestinal microbiota, reducing incidence of gastrointestinal infections, respiratory and also possessing a recognized bifidogenic effect. Traditionally, these prebiotic compounds have been obtained through extraction processes from some plants, as well as through enzymatic hydrolysis of sucrose. However, different fermentative methods have also been proposed for the production of fructooligosaccharides, such as solid-state fermentation utilizing various agroindustrial by-products. By optimizing the culture parameters, fructooligosaccharides yields and productivity can be improved. The use of immobilized enzymes and cells has also been proposed as being an effective and economic method for large-scale production of fructooligosaccharides. This paper is an overview on the results of recent studies on fructooligosacharides biosynthesis, physicochemical properties, sources, biotechnological production and applications.The authors thank the National Council of Science and Technology of Mexico (CONACYT) for funding this study. D. A. Flores-Maltos thank the CONACYT for the financial support provided for her postgraduate studies in the Food Science and Technology Program, Universidad Autonoma de Coahuila, Mexico

Partial purification and biochemical characterization of a new highly acidic NYSO laccase from Alcaligenes faecalis

Abstract Background Due to the multitude industrial applications of ligninolytic enzymes, their demands are increasing. Partial purification and intensive characterization of contemporary highly acidic laccase enzyme produced by an Egyptian local isolate designated Alcaligenes faecalis NYSO were studied in the present investigation. Results Alcaligenes faecalis NYSO laccase has been partially purified and intensively biochemically characterized. It was noticed that 40–60% ammonium sulfate saturation showed maximum activity. A protein band with an apparent molecular mass of ~ 50 kDa related to NYSO laccase was identified through SDS-PAGE and zymography. The partially purified enzyme exhibited maximum activity at 55 °C and pH suboptimal (2.5–5.0). Remarkable activation for enzyme activity was recognized after 10-min exposure to temperatures (T) 50, 60, and 70 °C; time elongation caused inactivation, where ~ 50% of activity was lost after a 7-h exposure to 60 °C. Some metal ions Cu2+, Zn2+, Co2+, Ni2+, Mn2+, Cd2+, Cr2+, and Mg2+ caused strong stimulation for enzyme activity, but Fe2+ and Hg2+ reduced the activity. One millimolar of chelating agents [ethylenediamine tetraacetic acid (EDTA), sodium citrate, and sodium oxalate] caused strong activation for enzyme activity. Sodium dodecyl sulfate (SDS), cysteine-HCl, dithiothreitol (DTT), β-mercaptoethanol, thioglycolic acid, and sodium azide caused strong inhibition for NYSO laccase activity even at low concentration. One millimolar of urea, imidazole, kojic acid, phenylmethylsulfonyl fluoride (PMSF), H2O2, and Triton X-100 caused activation. The partially purified NYSO laccase had decolorization activity towards different dyes such as congo red, crystal violet, methylene blue, fast green, basic fuchsin, bromophenol blue, malachite green, bromocresol purple eriochrome black T, and Coomassie Brilliant Blue R-250 with various degree of degradation. Also, it had a vast range of substrate specificity including lignin, but with high affinity towards p-anisidine. Conclusion The promising properties of the newly studied laccase enzyme from Alcaligenes faecalis NYSO strain would support several industries such as textile, food, and paper and open the possibility for commercial use in water treatment. It will also open the door to new applications due to its ligninolytic properties in the near future

Application of Experimental Designs for Optimization the Production of <em>Alcaligenes Faecalis</em> Nyso Laccase

713-722A sequential optimization strategy based on statistical experimental designs was implemented in order to enhance laccase production by a local isolate Alcaligenes faecalis NYSO in a submerged culture. To screen the parameters significantly influencing the laccase productivity, a 2-level Plackett-Burman design was applied. Among the studied variables, the pH, yeast extract, (NH4)2SO4, glucose, and CuSO4.5H2O were selected based on their high positive significant effect on laccase productivity. In order to find out the combination among the most significant variables that brings maximum yield, Response Surface Methodology was applied, where a 3-level Box-Behnken design was utilized to create a polynomial quadratic model correlating the relationship between the five variables and the laccase productivity. The optimal combination of the major medium constituents for laccase production was evaluated using the JMP program, was as follows: yeast extract, 0.896%; (NH4)2SO4, 0.035%; CuSO4.5H2O, 0.0075%; FeSO4.7H2O, 0.000133%; glucose, 0.0943%, pH 10.6 and 30 oC for 24 hrs. The predicted optimum laccase activity was 791U ml-1 min-1, which was 700 times the activity with basal medium. In addition, the further optimization for both pH, CuSO4.5H2O concentration lead the yield to be 2435 U ml-1min at pH 11.0, 200 mg CuSO4.5H2O which achieved after 18 hrs incubation time