The hybrid peptide cecropin A (1⁻8)⁻LL37 (17⁻30) (C⁻L), derived from the sequence of cecropin A (C) and LL-37 (L), showed significantly increased antibacterial activity and minimized hemolytic activity than C and L alone. To obtain high-level production of C⁻L, the deoxyribonucleic acid sequence encoding C⁻L with preferred codons was cloned into pET-SUMO to construct a fusion expression vector, and overexpressed in Escherichia coli (E. coli) BL21 (DE3). The maximum fusion protein (92% purity) was obtained with the yield of 89.14 mg/L fermentation culture after purification with Ni-NTA Sepharose column. The hybrid C⁻L was cleaved from the fusion protein by SUMO-protease, and 17.54 mg/L pure active C⁻L was obtained. Furthermore, the purified C⁻L showed identical antibacterial and hemolytic activity to synthesized C⁻L. Stability analysis results exhibited that the activity of C⁻L changed little below 80 °C for 20 min, but when the temperature exceeded 80 °C, a significant decrease was observed. Varying the pH from 5.0 to 10.0 did not appear to influence the activity of C⁻L, however, pH below 4.0 decreased the antibacterial activity of C⁻L rapidly. Under the challenge of several proteases (pepsin, trypsin, and proteinase K), the functional activity of C⁻L was maintained over 50%. In summary, this study not only supplied an effective approach for high-level production of hybrid peptide C⁻L, but paved the way for its further exploration in controlling infectious diseases of farm animals or even humans.