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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. The current study deals with bioethanol production from pretreated Saccharum spontaneum with employment of strain improvement of Saccharomyces cerevisiae. Alkaline pretreatment with 2 % NaOH resulted in increased cellulose content to 82 %. Optimal conditions for saccharification were found to has significantly (p 0.05) elevated saccharification rate with 149.47 ± 1.46 mg/ml release of glucose. It suggested that the utilization of feedstock (Saccharum spontaneum) under its optimum conditions of enzymatic hydrolysis can lead to the augmented sugars release which contributes significantly to the production of bioethanol with a future prospect of industrial exploitation.