Purification and identification of a novel antifungal protein from Bacillus subtilis XB-1
This study aimed to characterize a powerful antifungal component from bacteria. Bacillus subtilis strain XB-1, which showed maximal inhibition of Monilinia fructicola, was isolated and identified, and an antifungal protein was obtained from it. Ammonium sulfate precipitation, ion exchange chromatography, and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) were used to purify and identify the proteins secreted by B. subtilis XB-1. Analyses revealed that purified fraction V had the strongest antifungal effect, with the largest pathogen inhibition zone diameter of 4.15 cm after 4 days (P < 0.05). This fraction showed a single band with a molecular weight of approximately 43 kDa in SDS-PAGE. Results from SDS-PAGE and liquid chromatography electrospray ionization tandem mass spectrometry analyses demonstrated that fraction V was likely a member of the chitosanase family. These results suggest that B. subtilis XB-1 and its antifungal protein may be useful in potential biocontrol applications.
KeywordsAntifungal protein Bacillus subtilis Chitosanase Monilinia fructicola
This work was financially supported by the Science Foundation of Changzhou University (Grant No. ZMF18020316), Hebei Cixin Environmental Technology Co., Ltd, China (Grant No. 2018K0948), and Shaanxi Xintiandi Grass Industry Co., Ltd, China (Grant No. 2018K0947).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflicts of interest.
- Cao XH, Liao ZY, Wang CL, Yang WY, Lu MF (2009) Evaluation of a lipopeptide biosurfactant from Bacillus natto TK-1 as a potential source of anti-adhesive, antimicrobial and antitumor activities. Braz J Microbiol 40:373–379. https://doi.org/10.1590/S1517-838220090002000030 CrossRefPubMedPubMedCentralGoogle Scholar
- Casals C, Elmer PAG, Vinas I, Teixido N, Sisquella M, Usall J (2012) The combination of curing with either chitosan or Bacillus subtilis CPA-8 to control brown rot infections caused by Monilinia fructicola. Postharvest Biol Technol 64:126–132. https://doi.org/10.1016/j.postharvbio.2011.06.004 CrossRefGoogle Scholar
- Gell I, De Cal A, Torres R, Usall J, Melgarejo P (2008) Relationship between the incidence of latent infections caused by Monilinia spp. and the incidence of brown rot of peach fruit: factors affecting latent infection. Eur J Plant Pathol 121:487–498. https://doi.org/10.1007/s10658-008-9268-3 CrossRefGoogle Scholar
- Hernandez-Rodriguez A, Heydrich-Perez M, Acebo-Guerrero Y, Velazquez-del Valle MG, Hernandez-Lauzardo AN (2008) Antagonistic activity of Cuban native rhizobacteria against Fusarium verticillioides (Sacc.) Nirenb. in maize (Zea mays L.). Appl Soil Ecol 39:180–186. https://doi.org/10.1016/j.apsoil.2007.12.008 CrossRefGoogle Scholar
- Kilani-Feki O, Ben Khedher S, Dammak M, Kamoun A, Jabnoun-Khiareddine H, Daami-Remadi M, Tounsi S (2016) Improvement of antifungal metabolites production by Bacillus subtilis V26 for biocontrol of tomato postharvest disease. Biol Control 95:73–82. https://doi.org/10.1016/j.biocontrol.2016.01.005 CrossRefGoogle Scholar
- Kumar NSS, Nazeer RA, Jaiganesh R (2012) Purification and identification of antioxidant peptides from the skin protein hydrolysate of two marine fishes, horse mackerel (Magalaspis cordyla) and croaker (Otolithes ruber). Amino Acids 42:1641–1649. https://doi.org/10.1007/s00726-011-0858-6 CrossRefGoogle Scholar
- Liu J, Sui Y, Wisniewski M, Droby S, Tian SP, Norelli J, Hershkovitz V (2012) Effect of heat treatment on inhibition of Monilinia fructicola and induction of disease resistance in peach fruit. Postharvest Biol Technol 65:61–68. https://doi.org/10.1016/j.postharvbio.2011.11.002 CrossRefGoogle Scholar
- Srikhong P, Lertmongkonthum K, Sowanpreecha R, Rerngsamran P (2018) Bacillus sp. strain M10 as a potential biocontrol agent protecting chili pepper and tomato fruits from anthracnose disease caused by Colletotrichum capsici. Biocontrol 63:833–842. https://doi.org/10.1007/s10526-018-9902-8 CrossRefGoogle Scholar
- Tan TM, Zhu JX, Shen AR, Li JL, Yu YT, Zhang MJ, Zhao MR, Li ZM, Chen J, Gao CS, Cheng Y, Guo LT, Yan L, Sun XP, Zeng LB, Yan Z (2019) Isolation and identification of a Bacillus subtilis HZ-72 exhibiting biocontrol activity against flax seedling blight. Eur J Plant Pathol 153:825–836. https://doi.org/10.1007/s10658-018-1595-4 CrossRefGoogle Scholar