South Asian Journal of Research in Microbiology

The need for environmentally sustainable enzyme extraction methods has increased due to the growing demand for industrial bioprocesses. In this study, slime from Achatina achatina combined with syringe-needle hydraulic pressure was used to lyse Saccharomyces cerevisiae cells and recover a crude intracellular enzyme extract. Cells from a stock of a highly ethanol-tolerant strain of Saccharomyces cerevisiae isolated from palm wine were cultured on modified Potato Dextrose Agar (PDA) supplemented with 0.5% potassium phosphate (K₂HPO₄) and incubated in a candle jar for 3 days. A standardised yeast suspension was prepared from the resulting colonies. Equal volumes of Saccharomyces cerevisiae suspension and snail slime (1:1 ratio) were mixed and left to stand for 1 hour. The mixture was repeatedly homogenized by hydraulic pressure through repeated syringe-needle (340 µm) passage for three rounds. The lysed cell suspension was filtered through a 0.45 µm membrane filter to obtain a crude enzyme extract. The extract was evaluated for its ethanol production potential using glucose solutions at different concentrations (1%, 2.5%, 5%, 7.5%, and 10% v/v). The results showed that no ethanol was detected at 1% glucose, while ethanol yield increased progressively with increasing glucose concentration, from 0.27% v/v at 2.5% glucose concentration to 0.37% v/v at 10% glucose concentration. The absence of ethanol at 1% glucose likely reflected insufficient carbon substrate, whereas higher glucose levels supported measurable enzymatic activity. These findings indicated that snail slime-assisted lysis combined with hydraulic pressure effectively disrupted yeast cells and yielded active enzyme extracts capable of ethanol production. This approach offers a simple, eco-friendly, and low-cost alternative to conventional crude enzyme extraction techniques and may have potential applications in industrial biotechnology and cell-free ethanol production systems.