The invention discloses a bacillus subtilis gene recombinant strain. The bacillus subtilis is introduced with a heat-resistant sucrose phosphorylase gene, and that nucleotide sequence of the heat-resistant sucrose phosphorylase gene is shown in SEQ ID NO. 2, wherein the nucleotide sequence of the heat-resistant sucrose phosphorylase gene is shown in SEQ ID NO. The invention also discloses a methodfor preparing glycerol glucoside by using the heat-resistant sucrose phosphorylase biocatalysis, wherein the glycerol glucoside obtained is high in purity and low in cost, and lays a foundation for realizing large-scale biological preparation of glycerol glucoside.