This article presents a detailed study on the conditions for achieving a stable biocatalyst to be used in the production of ethanol from starch. Different pellets were used depending on which characteristic of the biocatalyst was being studied: (a) Saccharomyces cerevisiae entrapped in pectin or calcium alginate gel particles; (b) silica containing immobilized glucoamylase entrapped in pectin gel particles; or (c) pectin gel particles, with the silica-enzyme derivative and yeast coimmobilized. The influence of several variables on the mechanical resistance of the particle, on the viability of the microorganism, and on the rate of substrate hydrolysis was studied with biocatalyst. The best conditions found were 6% pectin gel, 2-mm particle diameter, and cure in 0.2 M CaCl2.2H(2)O/60 mM acetate buffer, pH 4.2, for gel preparation; and 6.0 g/L of CaCl2.2H(2)O in the fermentation medium. Biocatalyst (c) was successfully tasted for the production of ethanol from liquefied manioc flour syrup.