The esterase SulE is known to be an important and primary enzyme for the degradation of many sulfonylurea herbicides such as tribenumn-methyl, metsulfuron-methyl, and chlorimumn-ethyl. The disadvantages of applying free enzyme include poor stability and rapid deactivation in the natural environment, but these problems can be mitigated by using immobilized enzymes. In this study, cross-linked gelatin/chitosan (GLT/CTS) was selected as the enzyme immobilization carrier because its SulE activity recovery was higher than those of gamma-polyglutamate/gelatin (gamma-PGA/GLT) and gamma-polyglutamate/chitosan (gamma-PGA/CTS). The immobilization conditions were further optimized, and the effects of pH and temperature on GLT/CTS-SulE and free SulE activities to degrade tribenuron-methyl and metsulfuron-methyl were investigated. A microcosm experiment was conducted to assess the effectiveness of GLT/CTS-SulE in remediating soils polluted with tribenuron-methyl and metsulfumn-methyl. The results showed that, compared with free SulE, GLT/CTS-SulE displayed better pH and temperature adaptabilities and higher degradation efficiencies for both herbicides in soil, especially tribenuronmethyl. The results indicate that this is an excellent method to immobilize SulE for enhanced remediation of soils polluted with tribenuron-methyl or metsulfumn-methyl in situ.