Production and Characterisation of Low-Cost Amylase from Fermented Cassava Mash Using Saccharomyces cerevisiae and Lactobacillus sp. for Industrial Applications.

UNLABELLED: Cassava mash wastewater is a largely underutilised agro-industrial byproduct with potential as a sustainable substrate for enzyme production and waste valorisation. This study investigated the biochemical properties and potential applications of amylase produced via solid-state fermentation of cassava root wastewater using Saccharomyces cerevisiae and Lactobacillus sp. (SLL). The crude enzyme was extracted and purified through ammonium sulfate precipitation, ion-exchange, and gel-filtration chromatography. Purification increased enzyme purity 12.9-fold with a recovery yield of 17%. The partially purified amylase exhibited broad catalytic activity across a wide pH and temperature spectrum, with an optimum at 30 °C and pH 2. It retained 90-70% activity between pH 2-12 and 86-76% activity at 30-90 °C after 3 h, demonstrating exceptional stability. Activity was largely unaffected by Na⁺, K⁺, Ca2⁺, Mg2⁺, Cu2⁺, and Zn2⁺, but inhibited by Hg2⁺ at 10 mM. Strong inhibition was observed with SDS (10%) and mercaptoethanol (5-10%), while no significant inhibition was recorded with L-cysteine, sodium azide, Tween-20, or urea. Remarkably, polar organic solvents enhanced enzyme activity. Furthermore, the amylase displayed high compatibility with commercial detergents and effective stain removal, underlining its industrial relevance. Collectively, these findings highlight the enzyme's robustness, solvent tolerance, and detergent compatibility, positioning it as a promising candidate for detergent formulations and broader biotechnological applications. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12088-025-01527-3.