Hyperproduction of Manganese Peroxidase through Chemical Mutagenesis of Trametes versicolor IBL-04 and Optimization of Process Parameters

Abstract

This is the first report on chemical mutagenesis of Trametes versicolor IBL-04 to develop a hyper-producing mutant for overproduction of manganese peroxidase (MnP) using sugarcane bagasse as a substrate. A freshly prepared inoculum of indigenously isolated T. versicolor IBL-04 was treated with 100 µg mL -1 (v/v) ethyl methane sulfonate (EMS) and ethidium bromide (EB) separately for different time periods. The selected mutants and parent strain were cultured in solid-state fermentation (SSF) conditions to select the hyper-producing mutants. After selection of hyper-producing EMS-and EB-treated mutants, the fermentation parameters, including substrate type, incubation time, initial pH of the medium, temperature, moisture level, and carbon-to-nitrogen ratio (C:N), were optimized by adopting the Classical Optimization Strategy. T. versicolor IBL-04 treated for 90 min with EMS (EMS-90 mutant) gave maximum MnP production (935 U mL -1 ) after 8 days of fermentation. Supplementation with carbon and nitrogen sources significantly enhanced mutant growth, and under optimum conditions, the maximum MnP production by the mutant strain increased to 3045 U mL -1. The results indicated that the random chemical mutagenesis significantly enhanced the MnP production. The increased production of MnP by the EMS-90 mutant strain suggest its potential for commercial-scale enzyme production and biotechnological applications.

Studies on a better laccase-producing mutant of Fusarium incarnatum LD-3 under solid substrate tray fermentation

“…Khanam and Prasuna (2014) reported a 15% increase in the laccase production from P. cinnabarinus after mutagenesis with UV radiation, X-ray, ethidium bromide and colchicine. Chemical mutagen ethyl methanesulfonate was used to obtain better Mn peroxidaseproducing mutant of T. versicolor IBL-04 and EMS-90 mutant showed a 3.26-fold increase in the Mn peroxidase production (Ramzan et al 2013).…”

“…Improvement in the laccase production can also be achieved by mutating wild-type fungal strains followed by selection of the strain with improved character (Ramzan et al 2013). Application of UV radiation as a physical mutagen has been reported as one of the best methods of strain improvement for better productivity in mycelia cell and sporophore production in P. florida, P. pulmoniarus (Adegoke et al 2012) and P.cinnabarinus (Khanam and Gyanprasuna 2014).…”

Purification and biochemical characterization of extracellular manganese peroxidase from Ganoderma lucidum IBL-05 and its application

“…Triplicate conical flasks containing 5 g semi-solid wheat bran were autoclaved and inoculated with 5 ml homogenous inoculum (Ramzan et al, 2013). The inoculated flasks were allowed to ferment at 30°C in a temperature controlled incubator for 5 to 7 days at pH 4.5.…”

“…In this study, a locally isolated fungal strain, G. lucidum IBL -5was exploited for ligninolytic enzyme (Lip, MnP and laccase) production potential in solid state medium of wheat bran under pre-optimized growth conditions such as moisture, 50%; substrate, 5 g; pH, 5.5; temperature, 30°C; carbon source, 2% glucose; nitrogen source, 0.02% yeast extract; C:N ratio, 25:1; fungal spore suspension, 5 ml and fermentation time period, 5 days (Ramzan et al, 2013). The enzyme extract contained 576.3, 717.7 and 323.2 UmL -1 of LiP, MnP and laccase, respectively.…”

Beta-Glucosidase 1 (bgl1) Gene Analysis in Mutant and Wild-type of Penicillium sp. ID10-T065

“…The combined mutation of UV-EMS can induce a wide genetic variability (El-Bondkly & Keera, 2007). Several studies using a combination of UV light and EMS have proved the increase in the production of commercial enzymes, such as lipases (El-Bondkly & Keera, 2007); α-amylase (Shafique et al, 2009); phosphatase (Rajeshkumar & Ilyas, 2011); and ligninase (Ramzan et al, 2014).…”