The present relates to a method for generating variants of a protein based on a native protein regulated by allosteric pathway, the method comprising: - i) providing 3D structures of the native protein; - ii) identifying at least one pair of coupled allosteric sites within the amino acid sequence of the native protein named microswitch; - iii) generating in silico mutations of said identified microswitch to generate a pool of variants; - iv) computing at least one score reflecting the variation in allosteric coupling; and/or the variation in the relative stability - v) predicting the activity of each variant compared to the native protein based on the computed score. The invention also concern a computer implemented program to carry out said method, and a variant of a protein or an active fragment thereof, a polynucleotide.