Unstable oil markets with rising environmental concerns have revived widespread interest in production of biofuel from renewable materials. Cellulosic materials are abundant and prominent feedstocks for cost effective bioethanol production. A progressive move towards the sustainable and renewable energy sources has been started with the continuous depletion of conventional fossil fuels, rise in energy consumption and emission of greenhouse gas. Bioethanol is a proficient alternative fuel that can be biologically produced from waste materials and feedstock. In present investigation the production efficiency of thermotolerant and ethanol tolerant mutant strain was more than double as compared to parent strain was observed.