In this work, the production of fungal laccase was optimized from local isolate of Pleurotus ostreatus using solid state fermentation. Factorial design was used to study the effect of several nutrients on enzyme production. Purification and characterization of the enzyme and the effect of temperature, pH and gamma radiation on fungal growth and enzyme production was investigated. Optimization of production conditions yielded an enzyme with activity over 32,450 IU/g of fermented substrate. Factorial design was capable of establishing the conditions that multiplied the activity of the enzyme several folds, consequently, reducing the cost of production. The enzyme was capable of decolorizing several dyes with over 80% reduction in color confirming the aromatic degrading capability of laccase. The enzyme was also used in the synthesis of gold nanoparticles, proving that laccase from Pleurotus ostreatus has a strong potential in several industrial applications.

Enterococci are nosocomial pathogens that can form biofilms, which contribute to their virulence and antibiotic resistance. Although many genes involved in biofilm formation have been defined, their distribution among enterococci has not been comprehensively studied on a genome scale, and their diagnostic ability to predict biofilm phenotypes is not fully established. Here, we assessed the biofilm-forming ability of 90 enterococcal clinical isolates. Major patterns of virulence gene distribution in enterococcal genomes were identified, and the differentiating virulence genes were screened by polymerase chain reaction (PCR) in 31 of the clinical isolates. We found that detection of gelE in Enterococcus faecalis is not sufficient to predict gelatinase activity unless fsrAB, or fsrB alone, is PCR-positive (P = 0.0026 and 0.0012, respectively). We also found that agg is significantly enriched in isolates with medium and strong biofilm formation ability (P = 0.0026). Additionally, vancomycin, applied at sub minimal inhibitory concentrations, inhibited biofilm in four out of five strong biofilm-forming isolates. In conclusion, we suggest using agg and fsrB genes, together with the previously established gelE, for better prediction of biofilm strength and gelatinase activity, respectively. Future studies should explore the mechanism of biofilm inhibition by vancomycin and its possible use for antivirulence therapy.

Prodigiosin is a red pigment produced by Serratia marcescens. Prodigiosin is regarded as a promising drug owing to its reported characteristics of possessing anti-microbial, anti-cancer, and immunosuppressive activity. A factorial design was applied to generate a set of 32 experimental combinations to study the optimal conditions for pigment production using crude glycerol obtained from local biodiesel facility as carbon source for the growth of Serratia marcescens. The maximum production (870 unit/cell) was achieved at 22 °C, at pH 9 with the addition of 1% (w/v) peptone and 109 cell/ml inoculum size after 6 days of incubation. Gamma radiation at dose 200 Gy was capable of doubling the production of the pigment using the optimized conditions and manipulating production temperature. Our results indicate that we have designed an economic medium supporting enhanced Serratia marcescens MN5 prodigiosin production giving an added value for crude glycerol obtained from biodiesel industry.