Bioethanol production improvement by developing of ethanol-high tolerate Saccharomyces cerevisiae mutant and Taguchi statistical experimental-design method

Articles in Press

Document Type : Original Article- English

Authors

1 Department of biology

2 biology

10.22108/bjm.2026.148143.1667

Abstract

Saccharomyces cerevisiae is widely recognized as one of the most promising unicellular fungi, primarily due to its essential role in biotechnology and bioethanol production. As the global demand for sustainable energy sources grows, the need for cost-effective bioethanol production has become increasingly critical. Traditionally, the bioethanol industry relies on S. cerevisiae to ferment molasses as a carbon source. However, the rising cost of molasses has led to a continuous increase in production expenses, necessitating the development of alternative, more economical strategies. This study focuses on identifying innovative approaches to enhance bioethanol yield while simultaneously reducing production costs. To achieve this, a random mutagenesis technique using ethidium bromide exposure was employed to improve S. cerevisiae strains. Selection of the most efficient strains was carried out through adaptive evolution and evolutionary engineering, aiming to optimize fermentation performance and drive more cost-efficient bioethanol production. We developed the mutant strains which produced and selection was applied based on adaptive evolution and evolutionary engineering produce higher than 12.4±0.7% (v/v) alcohol whereas the early strains were unable to accept more than 10% (v/v) ethanol. The optimization of ethanol production was carried out using the Taguchi statistical experimental design method. Key medium components, including Brix molasses, ammonium sulfate, yeast extract, magnesium sulfate, and inoculation rate, were carefully adjusted to enhance fermentation efficiency. As a result, Saccharomyces cerevisiae ZF1 achieved a bioethanol yield of 13.25±0.50% (v/v) in flask cultures after medium optimization. Further scaling up in a 2-liter bioreactor with the optimized medium yielded 13.1±0.35% (v/v) and 13.40±0.50% (v/v) bioethanol, demonstrating its potential for industrial-scale molasses-based ethanol production. These findings highlight the effectiveness of the Taguchi method in optimizing fermentation conditions by significantly reducing the number of experimental trials and time required, ultimately improving efficiency in bioethanol production.

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Journal of Microbial Biology

Articles in Press, Accepted Manuscript
Available Online from 13 October 2026

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