Sporosarcina pasteurii(S. pasteurii) is a soil-derived Gram-positive bacterium that has been used to develop a variety of application scenarios using its efficient urease activity to induce calcium carbonate production. However, the problems of uncontrollable and unstable mineralization reactions still existed in the current mineralization application studies of S. pasteurii, which were limited by the lack of basic theoretical studies, especially lacking the research on the structure and expression regulation mechanism of urease genes which play a central role in mineralization. The further discovery of the urease gene information of S. pasteurii urease will not only help to analyze the regulation mechanism of urease expression, but also promote the application of S. pasteurii mineralization materials. Based on the above situation, this research first compared the growth and urease gene expression of S. pasteurii under three different culture conditions by transcriptome sequencing, and then predicted the urease gene structure of S. pasteurii by bioinformatics method, and then verified the prediction results by molecular biology method. Finally, it was found that the urease gene of S. pasteurii is a double operons structure. This paper reveals for the first time the double operons structure of S. pasteurii urease, which greatly advances the mechanistic study of S. pasteurii-induced mineralization and is a crucial step forward in the regulation of urease expression. The discovery of the double operons structure is also of great significance for genetic engineering studies of urease.