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Antimicrobial activity of a honeybee (Apis cerana) venom Kazal-type serine protease inhibitor
- Kim, Bo Yeon, Lee, Kwang Sik, Zou, Feng Ming, Wan, Hu, Choi, Yong Soo, Yoon, Hyung Joo, Kwon, Hyung Wook, Je, Yeon Ho, Jin, Byung Rae
- Toxicon 2013 v.76 pp. 110-117
- Apis cerana, Bacillus subtilis, Bacillus thuringiensis, Beauveria bassiana, Escherichia coli, Fusarium graminearum, Gram-positive bacteria, antibacterial properties, antifungal properties, cysteine, elastase, entomopathogenic fungi, honey bees, peptidase K, plasmin, proteinase inhibitors, subtilisin, t-plasminogen activator, threonine, thrombin, trypsin, venoms
- Insect-derived Kazal-type serine protease inhibitors exhibit thrombin, elastase, plasmin, proteinase K, or subtilisin A inhibition activity, but so far, no functional roles for bee-derived Kazal-type serine protease inhibitors have been identified. In this study, a bee (Apis cerana) venom Kazal-type serine protease inhibitor (AcKTSPI) that acts as a microbial serine protease inhibitor was identified. AcKTSPI contained a single Kazal domain that displayed six conserved cysteine residues and a P1 threonine residue. AcKTSPI was expressed in the venom gland and was present as a 10-kDa peptide in bee venom. Recombinant AcKTSPI Kazal domain (AcKTSPI-Kd) expressed in baculovirus-infected insect cells demonstrated inhibitory activity against subtilisin A (Ki 67.03 nM) and proteinase K (Ki 91.53 nM), but not against α-chymotrypsin or trypsin, which implies a role for AcKTSPI as a microbial serine protease inhibitor. However, AcKTSPI-Kd exhibited no detectable inhibitory effects on factor Xa, thrombin, tissue plasminogen activator, or elastase. Additionally, AcKTSPI-Kd bound directly to Bacillus subtilis, Bacillus thuringiensis, Beauveria bassiana, and Fusarium graminearum but not to Escherichia coli. Consistent with these findings, AcKTSPI-Kd showed antibacterial activity against Gram-positive bacteria and antifungal activity against both plant-pathogenic and entomopathogenic fungi. These findings constitute molecular evidence that AcKTSPI acts as an inhibitor of microbial serine proteases. This paper provides a novel view of the antimicrobial functions of a bee venom Kazal-type serine protease inhibitor.