No significant effect of interactions among variables was observed. The variables of Eq. (1) were determined by multiple regression analysis by the application of RSM. The overall linear regression equation showing the empirical relationship between laccase activity (Y) and four test variables in coded

units is represented by Eq. (2). equation(2) Y=1399.9+956(RH)+82.5(pH)+67.6(gramflour)−124(time) RAD001 solubility dmso Multiple regression model assumes a linear relationship between independent variable (RH, pH, gram flour, time) and dependent variable Y. It was observed that over incubation of the experimental setup for 20 days had a negative impact on laccase production. The goodness-of-fit of the model was checked by determining coefficient of determination (R2) and adjusted R2. The observed values of R2 explained that the fitted model could explain 97.6% of the total variation and hence proves the adequacy of model. The adjusted R2 corrects the R2 value for the sample size and for number of terms in the model. The adjusted R2 value (94.3%) in the present study shows the

high significance of the model. Previous SSF studies have shown low laccase production by different wood rotting fungi with increase in incubation time. 18 This may be attributed to the exhaustion in available nutrient HKI-272 cell line and gaseous exchange with progress of time. 17 Main effects graphs showed that basic pH is more significant than acidic pH for enzyme production. Previous studies have shown acidic conditions to be stimulatory for laccase production. It may due to the habitat from which fungal strains

have been isolated. Fungi growing in acidic environment come in contact with various acidic plant phenols or pesticides.19 However, efficient laccase production under both, acidic and alkaline conditions suggests Coriolus sp. as versatile source that can thrive and produce enzyme irrespective of environmental pH condition. Gram flour supplementation, good source of organic carbon and nitrogen, is also significant for laccase production (Eq. 2). Previous studies have shown nitrogen Rolziracetam supplementation to be an important component for laccase production with high C/N ratio. 19 C/N ratio of gram flour was 0.85. The total laccase activity reported is higher than most of the previous reports making this indigenous isolate a suitable strain for laccase production. The indigenous isolate Coriolus sp. was found to be one of the good laccase producers. SSF resulted in 8870 fold increase in laccase activity at RH 89%; gram flour 1 g/5 gds; pH 5.0 and 10 days of incubation, compared to SmF. This is the first report of the cumulative effect of bioprocess variables (pH, RH and incubation time) and alternative nitrogen source (gram flour) on laccase production using Coriolus sp. All authors have none to declare. We acknowledge Jaypee University of Information Technology for providing financial assistance for the project.