Synthesis of Biologically Active Gibberellins and Surface-Active Substances by Nocardia vaccinii IMV B-7405 in the Presence of Erythritol
DOI:
https://doi.org/10.15407/Keywords:
phytohormones, biosynthesis precursor, surfactants, antimicrobial activity, destruction of biofilms, phytopathogenic bacteriaAbstract
A promising direction today is the creation of integrated technologies for obtaining complex microbial preparations for crop production. The ability of Nocardia vaccinii IMV B-7405 to simultaneously synthesize surfactants with antimicrobial activity and stimulating phytohormones (auxins, cytokines, gibberellins) was previously established. However, the concentration of synthesized gibberellins was low. One of the approaches to the intensification of the target metabolites synthesis is the introduction of a precursor of their biosynthesis into the cultivation medium. Since in most bacteria gibberellins are formed in the methyl-erythritol-4-phosphate pathway, it was assumed that erythritol could be the precursor of the synthesis of these phytohormones. In addition, surfactants are secondary metabolites, the biological activity of which can vary depending on the cultivation conditions. The aim of the work is to investigate the possibility of increasing the synthesis of gibberellic phytohormones by the surfactant producer N. vaccinii IMV B-7405 in the presence of exogenous erythritol in the culture medium and to determine the effect of erythritol on the biological properties of surfactants synthesized under such conditions. Methods. Cultivation of N. vaccinii IMV B-7405 was carried out in a medium with refined or waste oil (2%, volume fraction) containing 100–500 mg/L erythritol. The concentration of gibberellins was determined by the method of high-performance liquid chromatography, while the concentration of surfactants - by the weight method after extraction and modification with a mixture of chloroform and methanol. The antimicrobial activity of surfactants against phytopathogenic bacteria was analyzed by the indicator of the minimum inhibitory concentration, and the degree of destruction of biofilms – by the spectrophotometric method. Results. It was established that the introduction of 300–400 mg/L erythritol into the culture medium of N. vaccinii IMV B-7405 was accompanied by the 2–14 times increase in the concentration of biologically active gibberellins GA3 and GA4 compared to the indicators of synthesis in the medium without the precursor. Data on the formation of gibberellins by N. vaccinii IMV B-7405 were correlated with the activity of one of the key enzymes of their biosynthesis: in the presence of erythritol, the activity of 2-C-methyl-D-erythritol-4-phosphate-cytidyltransferase was 3–12 times higher than in cells of the IMV B-7405 strain grown without a precursor. In the presence of erythritol, the formation of surfactants with high antimicrobial activity against phytopathogenic bacteria and the ability to destroy their biofilms were observed. The minimum inhibitory concentrations of surfactants synthesized by the addition of erythritol at the end of the exponential growth phase of the IMV B-7405 strain on waste oil for the pathogens of tomato bacteriosis were 0.8–25 μg/mL, that is, lower than the indicators established for surfactants formed without erythritol (3.13−100 μg/mL). Destruction of biofilms of phytopathogenic bacteria under the influence of surfactants obtained in the presence of erythritol in a medium with waste oil was 8–34% higher compared to destruction under the action of surface-active substances synthesized without this precursor. It was established that the introduction of erythritol into the culture medium of N. vaccinii IMV B-7405 was accompanied by a twofold increase in the activity of NADP+-dependent glutamate dehydrogenase (the key enzyme in the biosynthesis of aminolipids responsible for the antimicrobial activity of the surfactant complex). Conclusions. The obtained results are the basis for the development of a highly efficient integrated technology of biosynthesis of surface-active substances and phytohormones for crop production. The complex bacterial preparation can be used both to stimulate the growth of agricultural plants and to control the number of pathogens of their bacteriosis.
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References
Albermann, S., Elter, T., Teubner, A., Krischke, W., Hirth T., & Tudzynski, B. (2013). Characterization of novel mutants with an altered gibberellin spectrum in comparison to different wild-type strains of Fusarium fujikuroi. Applied Microbiology and Biotechnology, 97(17), 7779-7790. https://doi.org/10.1007/s00253-013-4917-7
Abdallah, D. B., Tounsi, S., Gharsallah, H., Hammami, A., & Frikha-Gargouri, O. (2018). Lipopeptides from Bacillus amyloliquefaciens strain 32a as promising biocontrol compounds against the plant pathogen Agrobacterium tumefaciens. Environmental Science and Pollution Research, 36, 36518−36529. https://doi.org/10.1007/s11356-018-3570-1
Al-Mutar, D. M. K., Noman, M., Alzawar, N. S. A., Qasim, H. H., Li, D., & Song, F. (2023). The extracellular lipopeptides and volatile organic compounds of Bacillus subtilis DHA41 display broad-spectrum antifungal activity against soil-borne phytopathogenic fungi. Journal of Fungi (Basel), 9(8), 797. https://doi.org/10.3390/jof9080797
Bligh, E. G., & Dyer, W. J. (1959). A rapid method for total lipid extraction and purification. Canadian Journal of Biochemistry and Physiology, 37(8), 911−917. https://doi.org/10.1139/y59-099
Bolivar-Anillo, H. J., González-Rodríguez, V. E., Cantoral, J. M., García-Sánchez, D., Collado, I. G., & Garrido, C. (2021). Endophytic bacteria Bacillus subtilis, isolated from Zea mays, as potential biocontrol agent against Botrytis cinerea. Biology (Basel), 10(6), 492. https://doi.org/10.3390/biology10060492
Bunsangiam, S., Thongpae, N., Limtong, S., & Srisuk, N. (2021). Large scale production of indole-3-acetic acid and evaluation of the inhibitory effect of indole-3-acetic acid on weed growth. Scientific Reports, 11(1), 13094. https://doi.org/10.1038/s41598-021-92305-w
Cao, Y., Pi, H., Chandrangsu, P., Li, Y., Wang, Y., Zhou, H., Xiong, H., Helmann, J. D., & Cai, Y. (2018). Antagonism of two plant-growth promoting Bacillus velezensis isolates against Ralstonia solanacearum and Fusarium oxysporum. Scientific Reports, 8(1), 4360. https://doi.org/10.1038/s41598-018-22782-z
Chen, L., Zhang, H., Zhao, S., Xiang, B., & Yao, Z. (2021). Lipopeptide production by Bacillus atrophaeus strain B44 and its biocontrol efficacy against cotton rhizoctoniosis. Biotechnology Letters, 43(6), 1183−1193. https://doi.org/10.1007/s10529-021-03114-0
Diamanti, E., Hamed, M. M., Lacour, A., Bravo, P., Illarionov, B., Fischer, M., Rottmann, M., Witschel, M., & Hirsch, A. K. H. (2022). Targeting the IspD enzyme in the MEP pathway: identification of a novel fragment class. ChemMedChem, 17(5), e202100679. https://doi.org/10.1002/cmdc.202100679
Ghadamgahi, F., Tarighi, S., Taheri, P., Saripella, G. V., Anzalone, A., Kalyandurg, P. B., Catara, V., Ortiz, R., & Vetukuri, R. R. (2022). Plant growth-promoting activity of Pseudomonas aeruginosa FG106 and its ability to act as a biocontrol agent against potato, tomato and taro pathogens. Biology (Basel), 11(1), 140. https://doi.org/10.3390/biology11010140
Gomes, M-Z. V., & Nitschke, M. (2012). Evaluation of rhamnolipids surfactants as agents to reduce the adhesion of Staphylococcus aureus to polystyrene surfaces. Letters in Applied Microbiology, 49(1), 960−965.
Hao, K., Ullah, H., Qin, X., Li, H., Li, F., & Guo, P. (2019). Efectiveness of Bacillus pumilus PDSLzg 1, an innovative hydrocarbon degrading bacterium conferring antifungal and plant growth promoting function. 3 Biotech, 9, 305. https://doi.org/10.1007/s13205-019-1842-1
Hazarika, D. J., Goswami, G., Gautom, T., Parveen, A., Das, P., Barooah, M., & Boro, R. C. (2019). Lipopeptide mediated biocontrol activity of endophytic Bacillus subtilis against fungal phytopathogens. BMC Microbiology, 19(1), 71. https://doi.org/10.1186/s12866-019-1440-8
Hernández Rodríguez, A., Díaz Pacheco, A., Martínez Tolibia, S. E., Melendez Xicohtencatl, Y., Granados Balbuena, S. Y., López y López, V. E. (2024). Bioprocess of gibberellic acid by Fusarium fujikuroi: The challenge of regulation, raw materials, and product yields. Journal of Fungi, 10(6), 418. https://doi.org/10.3390/jof10060418
Heuston, S., Begley, M., Gahan, C. G. & Hill, C. (2012). Isoprenoid biosynthesis in bacterial pathogens. Microbiology, 158(6), 1389-1401. https://doi.org/10.1099/mic.0.051599-0
Hossain, A., Islam Masum, M. M., Wu, X., Abdallah, Y., Ogunyemi, S. O., Wang, Y., Sun, G., Li, B., & An, Q. (2020). Screening of Bacillus strains in biocontrol of pathogen Dickeya dadantii causing stem and root rot disease of sweet potato. Biocontrol Science and Technology, 1-19. https://doi.org/10.1080/09583157.2020.1798356
Ke, X., Wu, Z., Liu, Y., Liang, Y., Du, M., & Li, Y. (2023). Isolation, antimicrobial effect and metabolite analysis of Bacillus amyloliquefaciens ZJLMBA1908 against citrus canker caused by Xanthomonas citri subsp. citri. Microorganisms, 11(12), 2928. https://doi.org/10.3390/microorganisms11122928
Kim, J., Le, K. D., Yu, N. H., Kim, J. I., Kim, J. C., & Lee, C. W. (2020). Structure and antifungal activity of pelgipeptins from Paenibacillus elgii against phytopathogenic fungi. Pesticide Biochemistry and Physiology, 163, 154−163. https://doi.org/10.1016/j.pestbp.2019.11.009
Kumar, J., Ramlal, A., Mallick, D., & Mishra, V. (2021). An overview of some biopesticides and their importance in plant protection for commercial acceptance. Plants (Basel), 10(6), 1185. https://doi.org/10.3390/plants10061185
Kumari, S., Prabha, C., Singh, A., Kumari, S., & Kiran, S. (2018). Optimization of indole-3-acetic acid production by diazotrophic B. subtilis DR2 (KP455653), isolated from rhizospere of Eragrostis cynosuroides. International Journal of Pharma Medicine and Biological Sciences, 7(2), 20-27. https://doi.org/10.18178/ijpmbs.7.2.20-27
Kuzuyama, T., Takagi, M., Kaneda, K., Dairi, T. & Seto, H. (2000). Formation of 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol from 2-C-methyl-D-erythritol 4-phosphate by 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase, a new enzyme in the nonmevalonate pathway. Tetrahedron Letters, 41, 703−706. https://doi.org/10.1016/S0040-4039(99)02143-7
Leonova, N. O., Pirog, Т. P., Piatetska, D. V., Shevchuk, T. A., Kharkhota, M. A., & Iutynska, G. O. (2020). Synthesis of gibberellins by surfactant producers Nocardia vaccinia IMV В-7405, Acinetobacter calcoaceticus IMV В-7241 and Rhodococcus erythropolis IMV Ас-5017. Scientific Study & Research: Chemistry & Chemical, 21(4),497-509.
Mazzola, P., Jozala, A., Lencastre-Novaes, L., Moriel, P., & Vessoni-Penna T. (2009). Minimal inhibitory concentration (MIC) determination of disinfectant and/or sterilizing agents. Brazilian Journal of Pharmaceutical Sciences, 45 (2), 241-248. https://doi.org/10.1590/S1984-82502009000200008
Myo, E. M., Ge, B., Ma, J., Cui, H., Liu, B., Shi, L., Jiang, M., & Zhang, K. (2019). Indole-3-acetic acid production by Streptomyces fradiae NKZ-259 and its formulation to enhance plant growth. BMC Microbiology, 19(1), 155. https://doi.org/10.1186/s12866-019-1528-1
Piatetska, D., & Pirog, T. (2023). Influence of Nocardia vaccinii IMV B-7405, Acinetobacter calcoaceticus IMV B-7241 and Rhodococcus erythropolis IMV Ac-5017 exometabolites on the harvest of tomato, pepper and barley. Scientific Works of NUFT, 29(5), 7−20. https://doi.org/10.24263/2225-2924-2023-29-5-3
Pirog, T. P., Lutsai, D. A., Shevchuk, T. A., & Iutynska, G. O. (2021а). Synthesis and biological activity of Аcinetobacter calcoaceticus IMV B-7241 surfactants depending on monovalent cations content in cultivation medium. Mikrobiolohichnyi Zhurnal, 83(2), 36−47. https://doi.org/10.15407/microbiolj83.02.020
Pirog, T., Piatetska, D., Leonova, N., & Shevchuk, T. (2024). Integrated technology of the surfactants and phytohormones biosynthesis by Nocardia vaccinii IMV B-7405 for their use in crop production. Ukrainian Food Journal, 13(1), 143-161. https://doi.org/10.24263/2304-974X-2024-13-1-10
Pirog, T. P., Piatetska, D. V., Yarova, H. А., & Iutynska, G. O. (2021b). Effect on phytopathogenic microorganisms of surfactants of microbial origin. Mikrobiolohichnyi Zhurnal, 83(6), 75−94. https://doi.org/10.15407/microbiolj83.06.075
Pirog, T., Sofilkanych, A., Konon, A., Shevchuk, T., & Ivanov, S. (2013). Intensification of surfactants' synthesis by Rhodococcus erythropolis IMV Ac-5017, Acinetobacter calcoaceticus IMV В-7241 and Nocardia vaccinii K-8 on fried oil and glycerol containing medium. Food and Bioproducts Processing, 91(2), 149−157. https://doi.org/10.1016/j.fbp.2013.01.001
Raajaraam, L., & Raman, K. A. (2022). Computational framework to identify metabolic engineering strategies for the co-production of metabolites. Frontiers in Bioengineering and Biotechnology, 9, 779405. https://doi.org/10.3389/fbioe.2021.779405
Rohmer, M., Knani, M., Simonin, P., Sutter, B., & Sahm, H. (1993). Isoprenoid biosynthesis in bacteria: a novel pathway for the early steps leading to isopentenyl diphosphate. Biochemical Journal, 295(Pt 2), 517-524. https://doi.org/10.1042/bj2950517
Salazar-Cerezo, S., Martínez-Montiel, N., García-Sánchez, J., Pérez-Y-Terrón, R., & Martínez-Contreras, R. D. (2018). Gibberellin biosynthesis and metabolism: a convergent route for plants, fungi and bacteria. Microbiology Research, 208, 85−98. https://doi.org/10.1016/j.micres.2018.01.010
Tao, H., Li, X., Huo, H., Cai, Y., & Cai, A. (2024). Bacillus velezensis Y6, a potential and efficient biocontrol agent in control of rice sheath blight caused by Rhizoctonia solani. Microorganisms, 12(8), 1694. https://doi.org/10.3390/microorganisms12081694
Wang, X., Liang, L., Shao, H., Ye, X., Yang, X., Chen, X., Shi, Y., Zhang, L., Xu, L., & Wang, J. (2022). Isolation of the novel strain Bacillus amyloliquefaciens F9 and identification of lipopeptide extract components responsible for activity against Xanthomonas citri subsp. citri. Plants (Basel), 11(3), 457. https://doi.org/10.3390/plants11030457
Wang, M., Zhang, Y., Cai, H., Zhao, X., Zhu, Z., Yan, Y., Yin, K., Cheng, G., Li, Y., Chen, G., Zou, L., & Tu, M. (2024). A new biocontrol agent Bacillus velezensis SF334 against rubber tree fungal leaf anthracnose and its genome analysis of versatile plant probiotic traits. Journal of Fungi (Basel), 10(2), 158. https://doi.org/10.3390/jof10020158
Yadav, A., Mudoi, K. D., Kumar, N., Geed, S. R., Gogoi, P., Sharma, R. K., & Saikia, R. (2022). Auxin biosynthesis by Microbacterium testaceum Y411 associated with orchid aerial roots and their efficacy in micropropagation. Frontiers in Plant Science, 13, 1037109. https://doi.org/10.3389/fpls.2022.1037109
Zhao, L., Chang, W. C., Xiao, Y., Liu, H. W., & Liu, P. (2013). Methylerythritol phosphate pathway of isoprenoid biosynthesis. Annual Review of Biochemistry, 82, 497−530. https://doi.org/10.1146/annurev-biochem-052010-100934
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