Study on the Antibacterial Activity of Secondary Metabolites from Lysobacter brunescens OH23 and Optimization of Fermentation Medium

doi:10.16409/j.cnki.2095-039x.2019.03.009

Abstract

Abstract: In this study, the antimicrobial spectrum of secondary metabolites of Lysobacter brunescens OH23 was tested. The results showed that OH23 was a potential biocontrol strain with specific activity against Xanthomonas sp. At the same time, the secondary metabolites of OH23 showed different antagonistic activity against different strains of Xanthomonas oryzae. Stability test indicated that the active secondary metabolites were stable with high temperature, ultraviolet light, protease and non-strong alkaline environment. However, the antagonistic activity almost completely lost under strong alkaline conditions (pH=11.0). Single factor test and orthogonal design were used to screen and optimize the carbon source, nitrogen source and metal ion of OH23 fermentation medium. The final formulation for high yield of active secondary metabolites included high polypeptone at 5 g/L, yeast powder at 5 g/L, soluble starch at 6 g/L, FeSO₄·7H₂O at 15 mg/L and ZnSO₄·7H₂O at 5 mg/L.

Key words: Lysobacter brunescens, Xanthomonas oryzae, secondary metabolites, antibacterial activity, medium optimization

CLC Number:

S476

ZHU Runjie, ZHAO Yancun, LING Jun, SONG Zhiwei, ZHAO Yangyang, XUE Xue, LIU Fengquan. Study on the Antibacterial Activity of Secondary Metabolites from Lysobacter brunescens OH23 and Optimization of Fermentation Medium[J]. journal1, 2019, 35(3): 426-436.

References

[1] Christensen P, Cook F. Lysobacter, a new genus of nonhiting, gliding bacteria with a high base ratio[J]. International Journal of Systematic Bacteriology, 1978, 28:367-393.
[2] Folman L, Postma J, van Veen J. Characterisation of Lysobacter enzymogenes (Christensen and Cook 1978) strain 3.1T8, a powerful antagonist of fungal diseases of cucumber[J]. Microbiological Rresearch, 2003, 158:107-115.
[3] Folman L, Klein M, Postma J, et al. Production of antifungal compounds by Lysobacter enzymogenes isolate 3.1T8 under different conditions in relation to its efficacy as a biocontrol agent of Pythium aphanidermatum in cucumber[J]. Biological Control, 2004, 31(2):145-154.
[4] Giesler L, Yuen G. Evaluation of Stenotrophomonas maltophilia strain C3 for biocontrol of brown patch disease[J]. Crop Protect, 1998, 17:509-513.
[5] Ko H, Jin R, Krishnan H, et al. Biocontrol ability of Lysobacter antibioticus HS124 against Phytophthora blight is mediated by the production of 4-hydroxyphenylacetic acid and several lytic enzymes[J]. Current Microbiology, 2009, 59:608-615.
[6] Kobayashi D Y, Reedy R M, Palumbo J D, et al. A clp gene homologue belonging to the Crp gene family globally regulates lytic enzyme production, antimicrobial activity, and biological control activity expressed by Lysobacter enzymogenes strain C3[J]. Applied and Environmental Microbiology, 2005, 71(1):261.
[7] Zhang W, Li Y, Qian G, et al. Identification and characterization of the anti-methicillin-resistant Staphylococcus aureus WAP-8294A2 biosynthetic gene cluster from Lysobacter enzymogenes OH11[J]. Antimicrob Agents Chemother, 2011, 55(12):5581-5589.
[8] Zhao Y, Qian G, Ye Y, et al. Heterocyclic aromatic N-oxidation in the biosynthesis of phenazine antibiotics from Lysobacter antibioticus[J]. Organic Letters, 2016, 18(10):2495-2498.
[9] Puopolo G, Cimmino A, Palmieri M C, et al. Lysobacter capsici AZ78 produces cyclo(L-Pro-L-Tyr), a 2,5-diketopiperazine with toxic activity against sporangia of Phytophthora infestans and Plasmopara viticola[J]. Journal of Applied Microbiology, 2015, 117(4):1168-1180.
[10] O'Sullivan J, McCullough J, Tymiak A, et al. Lysobactin, a novel antibacterial agent produced by Lysobacter sp. I. Taxonomy, isolation and partial characterization[J]. Journal of Antibiotics, 1988, 41:1740-1744.
[11] Bonner D, Osullivan J, Tanaka S, et al. Lysobactin, a novel antibacterial agent produced by Lysobacter sp. Ⅱ. Biological properties[J]. Journal of Antibiotics, 1988, 41:1745-1751.
[12] Hashizume H, Igarashi M, Hattori S, et al. Tripropeptins, novel antimicrobial agents produced by Lysobacter sp. I. Taxonomy, isolation and biological activities[J]. Journal of Antibiotics, 2001, 54(12):1054-1059.
[13] Hashizume H, Hattori S, Igarashi M, et al. Tripropeptin E (I), a new tripropeptin group antibiotic produced by Lysobacter sp. BMK333-48F3[J]. Journal of Antibiotics, 2004, 57(6):394.
[14] Hashizume H, Sawa R, Harada S, et al. Tripropeptin C blocks the lipid cycle of cell wall biosynthesis by complex formation with undecaprenyl pyrophosphate[J]. Antimicrob Agents Chemother, 2011, 55(8):3821-3828.
[15] Sangnoi Y, Anantapong T, Kanjanaopas A. Antibacterial activity of aquatic gliding bacteria[J]. Springerplus, 2016, 5(1):116.
[16] 张越, 张琦, 贺润平, 等. 植物病害防控方向和措施的研究探讨[J]. 山西农业科学, 2015, 43(1):75-78.
[17] 孔祥睿. 化学农药对环境的污染及其防控对策建议[J]. 南方农药, 2017(6):32-34.
[18] 杜士杰. 微生物源农药的应用现状及发展研究W[J]. 科学技术创新, 2016(25):155-155.
[19] 束长龙, 曹蓓蓓, 袁善奎, 等. 微生物农药管理现状与展望[J]. 中国生物防治学报, 2017, 33(3):297-303.
[20] 符泽, 邱光. 微生物农药的研发推广及未来发展趋势[J]. 农药市场信息, 2018(6):6-10.
[21] 孙洋, 王树桐, 曹克强. 影响生防菌田间药效环境因素的研究进展[C]//粮食安全与植保科技创新, 2009.
[22] 田囡. 产酶溶杆菌C3产生热稳定性抗真菌因子(HSAF)的研究[D]. 山东轻工业学院, 2011, 62-65.
[23] 张孟, 王艳婷, 汪立平, 等. 海洋抑菌放线菌Y15的筛选、鉴定及其代谢产物理化性质[J]. 微生物学通报, 2017, 44(3):513-524.
[24] 畅涛, 冯中红, 杨成德, 等. 一株抗马铃薯坏疽病生防细菌B-401产抑菌物质条件优化及稳定性[J]. 中国生物防治学报, 2016, 32(3):365-371.
[25] 章四平, 刘圣明, 王建新, 等. 枯草芽孢杆菌生防菌株NJ-18的发酵条件优化[J]. 南京农业大学学报, 2010, 33(2):58-62.
[26] 王云霞, 钱国良, 胡白石, 等. 产酶溶杆菌OH11菌株摇瓶发酵条件研究[J]. 中国生物防治学报, 2008, 24(3):267-271.
[27] 王亚娇, 纪莉景, 栗秋生, 等. 列当生防菌层出镰刀菌Br-1发酵条件的优化及田间防治效果[J]. 中国生物防治学报, 2017, 33(5):692-698.
[28] 乔俊卿, 刘邮洲, 张荣胜, 等. 枯草芽胞杆菌T-500产脂肽类抗生素的摇瓶发酵工艺优化[J]. 南京农业大学学报, 2018, 41(3):447-455.