This was the continuation of our previous study along the same line with more focus on technical details because the data are usually divided into two datasets, one for model development and the other for model validation during the development of predictive model. The widely used validation method is the delete-1 jackknife validation. However, no systematical studies were conducted to determine whether the jackknife validation with different deletions works better because the number of validations with different deletions increases in a factorial fashion. Therefore it is only small dataset that can be used for such an exhausted study. Cellulase is an enzyme playing an important role in modern industry, and many parameters related to cellulase in enzymatic reactions were poorly documented. With increased interests in cellulases in bio-fuel industry, the prediction of parameters in enzymatic reactions is listed on agenda. In this study, two aims were defined (a) which amino acid property works better to predict the temperature optimum and (b) with which deletion the jackknife validation works. The results showed that the amino acid distribution probability works better in predicting the optimum temperature of catalytic reaction by cellulase, and the delete-4, more precisely one-fifth deletion, jackknife validation works better.