On the basis of modeling analysis by software, the traditional deglycosylation way was changed while new N-glycosylation with enhanced aromatic sequences (EAS) was introduced into the reverse turn of PGL through semi-rational design.In consequence, the thermostability of alkaline polygalacturonate lyase (PGL) was improved and a PGL engineering bacteria with higher quality was obtained.Removing the N-glycosylation in N353 by replacing the threonine at site 355 with tryptophan that has low mutation energy raised the half-life of PGL by 177%, which was still far from the target yet.Then, introducing new N-glycosylation in site 127 and 137 brought the half-life of recombinase PGL up to 1.35 h that was close to the target value.Introducing N-glycosylation with EAS into the reverse turn of enzyme could play a significant role in improving its thermostability.