Activation of the Purine Salvage Pathway Increases the Production of Toyocamycin in Streptomyces diastatochromogenes 1628

doi:10.16409/j.cnki.2095-039x.2023.02.038

Abstract

Abstract: Toyocamycin is one kind of the nucleoside antibiotics, which is a promising fungicide to control the plant fungal pathogens. Streptomyces diastatochromogenes 1628 was shown to produce the toyocamycin, but the low yield of toyocamycin still limited the further industrial process. To increase the toyocamycin production, the transcriptome indicated that the purine salvage pathway affected the toyocamycin cluster and the gene xdhR in the purine salvage pathway was identified by homologue blast in the genome of S. diastatochromogenes 1628. Therefore, the functions of xdhR in toyocamycin production, transcription level of toy cluster and morphological differentiation were explored in this study. It was observed that the deletion of xdhR gene could significantly enhance the yield of toyocamycin of S. diastatochromogenes 1628, and the highest yield of toyocamycin in the recombinant strain reached 287.8 mg/L, which was about 114.0% higher than that of the wild type in 96 hours. The deletion of xdhR gene could also activate the transcription of purine salvage pathway genes and the genes in toy cluster. In addition, xdhR gene could change the morphological differentiation. The inhibition ratios of 1628_ΔxdhR fermentation broth on Rhizoctonia solani and Fusarium oxysporum f. sp. cucumerinum increased by 69.3% and 100%, respectively, compared with that of the wild type. The results show that XdhR involved in the synthesis of toyocamycin in Streptomyces diastatochromogenes 1628, which provide a theoretical basis for increasing toyocamycin titer in other biocontrol strains.

Key words: toyocamycin, Streptomyces, purine salvage pathway, biological pesticide

CLC Number:

S476

ZHANG Zixuan, SONG Yang, SHENTU Xuping, YU Xiaoping. Activation of the Purine Salvage Pathway Increases the Production of Toyocamycin in Streptomyces diastatochromogenes 1628[J]. Chinese Journal of Biological Control, 2023, 39(4): 851-860.

References

[1] Suhadolnik R J, Uematsu T. Biosynthesis of the pyrrolopyrimidine nucleoside antibiotic, toyocamycin:VII. Origin of the pyrrolecarbons and the cyano[J]. Journal of Biological Chemistry, 1970, 245(17):4365-4371.
[2] Ding Y, An H, Hong Z, et al. Synthesis of 2'-β-C-methyl toyocamycin and sangivamycin analogues as potential HCV inhibitors[J]. Bioorganic and Medicinal Chemistry Letters, 2005, 15(3):725-727.
[3] 申屠旭萍, 于冰, 边亚琳, 等. 拮抗链霉菌B28的分类鉴定及其生防作用研究[J]. 植物病理学报, 2012, 42(1):105-109.
[4] Latoscha A, Wörmann M E, Tschowri N. Nucleotide second messengers in Streptomyces[J]. Microbiology, 2019, 165(11):1153-1165.
[5] Shentu X, Liu N, Tang G, et al. Improved antibiotic production and silent gene activation in Streptomyces diastatochromogenes by ribosome engineering[J]. The Journal of Antibiotics, 2016, 69(5):406-410.
[6] McCarty R M, Bandarian V. Deciphering deazapurine biosynthesis:pathway for pyrrolopyrimidine nucleosides toyocamycin and sangivamycin[J]. Chemistry and Biology, 2008, 15(8):790-798.
[7] Kriel A, Bittner A N, Kim S H, et al. Direct regulation of GTP homeostasis by (p)ppGpp:a critical component of viability and stress resistance[J]. Molecular Cell, 2012, 48(2):231-241.
[8] Sivapragasam S, Grove A. Streptomyces coelicolor XdhR is a direct target of (p) ppGpp that controls expression of genes encoding xanthine dehydrogenase to promote purine salvage[J]. Molecular Microbiology, 2016, 100(4):701-718.
[9] 徐洁. GTP的合成对淀粉酶产色链霉菌1628产丰加霉素的影响[D]. 杭州:中国计量大学, 2020.
[10] Sivapragasam S, Grove A. The link between purine metabolism and production of antibiotics in Streptomyces[J]. Antibiotics, 2019, 8(2):76.
[11] Fu J, Zong G, Zhang P, et al. XdhR negatively regulates actinorhodin biosynthesis in Streptomyces coelicolor M145[J]. FEMS Microbiology Letters, 2017, 364(22):226.
[12] Wang J, Xu J, Luo S, et al. AdpA_sd, a positive regulator for morphological development and toyocamycin biosynthesis in Streptomyces diastatochromogenes 1628[J]. Current Microbiology, 2018, 75(10):1345-1351.
[13] Ihara Y, Ohta H, Masuda S. A highly sensitive quantification method for the accumulation of alarmone ppGpp in Arabidopsis thaliana using UPLC-ESI-qMS/MS[J]. Journal of Plant Research, 2015, 128(3):511-518.
[14] Werten S, Waack P, Palm G J, et al. Crystal structures of free and ligand-bound forms of the TetR/AcrR-like regulator SCO3201 from Streptomyces coelicolor suggest a novel allosteric mechanism[J]. The FEBS Journal, 2023, 290(2):521-532.
[15] Xu Y, Ke M, Li J, et al. TetR-type regulator SLCG_2919 is a negative regulator of lincomycin biosynthesis in Streptomyces lincolnensis[J]. Applied and Environmental Microbiology, 2019, 85(1):e02091-18.
[16] Lyu M, Cheng Y, Han X, et al. AccR, a TetR family transcriptional repressor, coordinates short-chain acyl coenzyme A homeostasis in Streptomyces avermitilis[J]. Applied and Environmental Microbiology, 2020, 86(12):e00508-20.
[17] Zhang J, Liang Q, Xu Z, et al. The inhibition of antibiotic production in Streptomyces coelicolor over-expressing the TetR regulator SCO3201 is correlated with changes in the lipidome of the strain[J]. Frontiers in Microbiology, 2020, 11:1399.
[18] Xia H, Zhan X, Mao X M, et al. The regulatory cascades of antibiotic production in Streptomyces[J]. World Journal of Microbiology and Biotechnology, 2020, 36(1):1-9.
[19] Fernández-Justel D, Marcos-Alcalde Í, Abascal F, et al. Diversity of mechanisms to control bacterial GTP homeostasis by the mutually exclusive binding of adenine and guanine nucleotides to IMP dehydrogenase[J]. Protein Science, 2022, 31(5):e4314.
[20] Zhu M, Dai X. Growth suppression by altered (p) ppGpp levels results from non-optimal resource allocation in Escherichia coli[J]. Nucleic acids Research, 2019, 47(9):4684-4693.
[21] 杨金红. 丝核菌属真菌病害生物防治研究进展[J]. 湖北农业科学, 2009, 48(10):2581-2584.
[24] 于冰, 申屠旭萍, 俞晓平. 丰加霉素对黄瓜立枯丝核菌的拮抗作用[J]. 中国生物防治学报, 2011, 27(3):373-377.
[22] 王俊博, 孔令新, 乔永健, 等. 基于基因组挖掘的农抗120活性成分制霉菌素和丰加霉素的发现和鉴定[J]. 微生物学报, 2021, 61(10):3076-3086.
[23] 朱昌雄, 田云龙, 蒋细良, 等. 4%增效农抗120对几种主要果树病害的试验效果[J]. 中国生物防治, 2004, 29(3):190-192.