Strain Improvement Through UV and Chemical Mutagenesis for Enhanced Citric Acid Production in Molasses-Based Solid State Fermentation

Abstract

Aspergillus niger was subjected to UV radiation and chemical mutagenesis to develop its hyper-producing mutants for enhanced citric acid production. Ethyl methane sulfonate (EMS) and Ethidium bromide (EB) were used for chemical mutagenesis of Aspergillus niger. UV, and chemically treated mutants of Aspergillus niger were identified by using 2-deoxy, D-glucose as selective marker. The selected mutants were cultured in solid state fermentation (SSF) of sugarcane molasses medium (10%) using corn cobs, banana stalk, sugarcane bagasse, wheat straw, and wheat bran as carrier substrates. After pH adjustment and sterilization, the triplicate flasks were inoculated with 5 mLof homogenous spore suspensions of selected mutants of A. niger and the flasks were subjected to SSF under still culture conditions. The mutant EB-3 (treated with 1 mg/mL ethidium bromide for 120 min) giving highest citric acid yield (64.2 mg/mL) in 72 h was selected as hyper-producing mutant. Citric acid production process using EB-3 mutant was then optimized to enhance citric acid production by the mutant in SSF. Aspergillus niger EB-3 mutant could produce 67.72 mg/mL citric acid in 72 h using banana stalks as support material under optimum conditions of pH (pH 6), incubation temperature (35°C) and inoculum size (5 mL) in SSF.