Severe acute respiratory syndrome coronavirus (SARS-CoV) is a highly pathogenic virus which originated from bat SARS-like coronavirus (SL-CoV) and infects humans. Angiotensin-converting enzyme 2 (ACE2), the functional receptor of SARS-CoV, is a key molecular affecting SARS-CoV pathogenesis, host range, and cross-species transmission. Previous studies demonstrated that SARS-CoV and some SL-CoV strains (such as WIV1) can use human, civet, and bat ACE2 to enter cells, while SARS-CoV utilizes mouse ACE2 less effectively. Though rodents are the most diverse mammals and the most popular experimental model, the utilization of mouse ACE2 by SL-CoV has not been revealed. In the present study, pseudoviral infection experiments were conducted to compare SARS-CoV BJ01 and SL-CoV WIV1 on their ability of using human, civet, bat, mouse ACE2 and ACE2 mutants to enter cells. Meanwhile, the binding between receptor-binding domain (RBD) of the two viruses and different ACE2 was verified by protein binding experiments. The results showed that SL-CoV WIV1 can use mouse ACE2 to enter cells, and WIV1 RBD binds to mouse ACE2 as efficiently as human and civet ACE2 whereas more strongly than bat ACE2. The L440P mutation of ACE2 in different species can significantly reduce the ability of RBD binding and the efficiency of pseudovirus invasion. The studies indicate that mouse ACE2 is a functional receptor for SL-CoV WIV1 and L440 of ACE2 is a key amino acid site affecting interspecies infection. The study is helpful to further understand the receptor recognition and cross-species transmission mechanism of SL-CoV, and will be beneficial to the prevention and control of similar CoV in the future.