Screening and Mutagenesis of Broad-Spectrum Antagonistic Bacillus licheniformis and Purification and Identification of Antimicrobial Substances Produced by Its Mutant

被引：2

作者：

Gao Z. ^{[1
]}

Wang Q. ^{[2
]}

Ding F. ^{[1
]}

Xu X. ^{[1
]}

Zhao Y. ^{[3
]}

Jiao W. ^{[4
]}

Chen T. ^{[1
]}

机构：

[1] College of Food (Biological) Engineering, Xuzhou Institute of Technology, Xuzhou

[2] Haibin Forest Farm, Qinhuangdao

[3] Yangtze River Guiliu Food Suining Co. Ltd., Xuzhou

[4] Pizhou Golden Earth Fertilizer Co. Ltd., Xuzhou

来源：

Shipin Kexue/Food Science | 2021年 / 42卷 / 02期

关键词：

Antifungal activity; Bacillus licheniformis; Lipopeptides; Mutagenesis; Purification;

D O I：

10.7506/spkx1002-6630-20191112-161

中图分类号：

学科分类号：

摘要：

The aims of this study were (1) to isolate and characterize bacterial strains capable of producing broad-spectrum antimicrobial substances and (2) to separate and identify these antimicrobial compounds. Strain identification was performed based on morphology, physiological and biochemical characteristics and 16S rDNA sequence analysis. To obtain genetically stable strains able to produce a high yield of antimicrobial compounds, we carried out mutagenesis by ultraviolet irradiation combined with diethyl sulfate treatment. Then, the antimicrobial substances were isolated and purified sequentially using methyl alcohol extraction, gel filtration chromatography (Sephadex G-10) and semi preparative high performance liquid chromatography (HPLC), and were identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). In this study, a total of 67 Bacillus strains were isolated from soils, eight of which showed strong antimicrobial effects, including strains XF32, which showed significant broad-spectrum antimicrobial and was identified as B. licheniformis. It displayed an antibacterial activity against seven pathogenic bacterial species, including Staphylococcus aureus, B. subtilis, B. cereus, Escherichia coli, Salmonella sp., Shigella sp., and Pseudomonas aeruginosa. The strain also exhibited antifungal activity against Saccharomyces cerevisiae, Aspergillus niger, Trichoderma atroviride and Fusarium oxysporum. Finally, mutant XF32-22 was obtained, whose antibacterial activity against S. aureus was increased significantly as compared to that of the original strain (XF32). Then, one antimicrobial substance was isolated and purified from the fermentation broth of XF32-22, and was identified as fengycin, a lipopeptide. The mutant produced significantly more fengycin than did its parental strain. Fengycin could strongly inhibit multiple bacterial pathogens as well as plant pathogenic fungi. Mutant XF32-22 holds great promise for application in food preservation and agricultural biological control. © 2021, China Food Publishing Company. All right reserved.

引用

页码：143 / 150

页数：7

共 27 条

[1]

INES M, DHOUHA G., Lipopeptide surfactants: production, recovery and pore forming capacity, Peptides, 71, pp. 100-112, (2015)

[2]

AYED H B, NASRI R, JEMIL N, Et al., Acute and sub-chronic oral toxicity profiles of lipopeptides from Bacillus mojavensis A21 and evaluation of their in vitro anticoagulant activity, Chemico-Biological Interactions, 236, pp. 1-6, (2015)

[3]

ONGENA M, JOURDAN E, ADAM A, Et al., Surfactin and fengycin lipopeptides of Bacillus subtilis as elicitors of induced systemic resistance in plants, Environmental Microbiology, 9, 4, pp. 1084-1090, (2007)

[4]

YU G Y, SINCLAIR J B, HARTMAN G L, Et al., Production of iturin A by Bacillus amyloliquefaciens suppressing Rhizoctonia solani, Soil Biology and Biochemistry, 34, 7, pp. 955-963, (2002)

[5]

SARWAR A, HASSAN M N, IMRAN M, Et al., Biocontrol activity of surfactin A purified from Bacillus NH-100 and NH-217 against rice bakanae disease, Microbiological Research, 209, pp. 1-13, (2018)

[6]

ROMERO D, DE VICENTE A, RAKOTOALY R H, Et al., The iturin and fengycin families of lipopeptides are key factors in antagonism of Bacillus subtilis toward Podosphaera fusca, Molecular Plant-Microbe Interactions, 20, 4, pp. 430-440, (2007)

[7]

BALAN S S, KUMAR C G, JAYALAKSHMI S., Pontifactin, a new lipopeptide biosurfactant produced by a marine Pontibacter korlensis strain SBK-47: purification, characterization and its biological evaluation, Process Biochemistry, 51, 12, pp. 2198-2207, (2016)

[8]

JIN P, WANG H, LIU W, Et al., A new cyclic lipopeptide isolated from Bacillus amyloliquefaciens HAB-2 and safety evaluation, Pesticide Biochemistry and Physiology, 147, pp. 40-45, (2018)

[9]

JHA S S, JOSHI S J, GEETHA S J., Lipopeptide production by Bacillus subtilis R1 and its possible applications, Brazilian Journal of Microbiology, 47, 4, pp. 955-964, (2016)

[10]

BANAT I M, SATPUTE S K, CAMEOTRA S S, Et al., Cost effective technologies and renewable substrates for biosurfactants' production, Frontiers in Microbiology, 5, (2014)

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