Identifying influential factors in the bacterial leaching of nickel laterites using a mixed culture of chemolithotrophic micro-organisms was explored using the approach of statistical design of experiments. In a series of experiments, pH, particle size, pulp density, type of substrate and inoculum size were statistically combined using a quarter fractional factorial designs 2 5−2 III and tested for their influence on nickel recovery using chemolithotrophic microorganisms. The results indicated that inoculum size was not statistically significant while the rest of the factors were statistically significant. Under the ranges studied the interaction between the variables was found to be weak. The results also showed that recovery was maximized at low pH and low pulp density. In the range studied, particles of less than 38μm had a negative influence on nickel recovery. Sulphur substrate also showed better effects than pyrite.