Thurincin H Is a Nonhemolytic Bacteriocin of Bacillus thuringiensis with Potential for Applied Use.

Thurincin H, a bacteriocin produced by Bacillus thuringiensis, exhibits antibacterial activity against Gram-positive and Gram-negative bacteria. While much is known about its expression and antimicrobial spectrum, its hemolytic property has yet to be established. In this study, thurincin H was produced in a plasmid-free acrystalliferous strain of B. thuringiensis (Bt CryB) that naturally lacked antimicrobial and hemolytic activities. When grown in Tryptic Soy Broth (TSB), the bacteriocin's maximal production in Bt CryB harboring the thurincin H genetic cluster (Bt CryB/pThur) was observed at 24 h. Thurincin H was purified as a sole peptide of ~5 kDa using three purification steps, i.e., salt precipitation, ultrafiltration, and gel filtration chromatography. The bacteriocin showed inhibitory activity against B. cereus (5631 U), Bt CryB (8827 U), E. faecium wild type (11,197 U), and E. faecium ATCC 19,434 (6950 U), but not against Bt CryB/pThurH and Bt CryB/pThurHΔThnA. In addition, a minimum inhibitory concentration (MIC) of 5.0 μg/mL against B. cereus 183 was observed. In silico predictions suggested that thuricin H lacks hemolytic activity, which was validated in vitro using 4 × the MIC, i.e., 20 μg/ml. Our data lay a foundation for the potential safe use of thurincin H as an antibacterial peptide for medical use, in food products, and for expression in probiotic bacteria.