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.