激活嘌呤回补途径提高淀粉酶产色链霉菌丰加霉素产量

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

中国生物防治学报 ›› 2023, Vol. 39 ›› Issue (4): 851-860.DOI: 10.16409/j.cnki.2095-039x.2023.02.038

激活嘌呤回补途径提高淀粉酶产色链霉菌丰加霉素产量

张子轩, 宋阳, 申屠旭萍, 俞晓平

中国计量大学生命科学学院/浙江省生物计量及检验检疫技术重点实验室, 杭州 310018

收稿日期:2022-11-16 出版日期:2023-08-25 发布日期:2023-08-25
通讯作者: 宋阳, 博士, 讲师, E-mail:songyang@cjlu.edu.cn;俞晓平, 博士, 研究员, E-mail:yxp@cjlu.edu.cn
作者简介:张子轩，硕士研究生，E-mail：511811129@qq.com。
基金资助:
浙江省自然科学基金（LGN22C140006）；国家自然科学基金（31901930）；浙江省属高校基本科研业务费专项资金；浙江省“三农九方”科技协作计划（2023SNJF034）；浙江省“领雁”研发攻关计划项目（2023C02030）

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

ZHANG Zixuan, SONG Yang, SHENTU Xuping, YU Xiaoping

Zhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine/College of Life Science, China Jiliang University, Hangzhou 310018, China

Received:2022-11-16 Online:2023-08-25 Published:2023-08-25

摘要/Abstract

摘要： 丰加霉素是核苷类抗生素家族中的一员,它在抑制真菌方面表现出很高的活性,在植物病害生物防治领域具有重要的应用潜力。淀粉酶产色链霉菌Streptomyces diastatochromogenes 1628是丰加霉素的生产菌株之一,但由于野生菌产量较低,难以实现在工业水平进行丰加霉素的发酵生产。通过对代谢途径分析发现,嘌呤回补途径能够影响丰加霉素合成。为了阐明嘌呤回补途径调控基因xdhR对淀粉酶产色链霉菌丰加霉素合成的影响,本研究通过基因敲除技术探究xdhR基因对丰加霉素产量、丰加霉素合成基因簇转录水平、菌丝分化、抑菌效果等方面的影响。摇瓶发酵试验表明,敲除xdhR基因能够显著提高淀粉酶产色链霉菌1628的丰加霉素产量,发酵至96 h时,重组菌中丰加霉素产量达到最高287.8 mg/L,产量较野生菌株提高约114.0%。qPCR试验表明,xdhR基因能够激活嘌呤回补途径基因的转录和丰加霉素合成基因簇基因的转录。同时,xdhR基因能够改变菌落生长形态。xdhR基因敲除菌株发酵液对水稻纹枯病菌和黄瓜枯萎病菌的抑制率分别提高到69.3%和100%。以上结果证明XdhR参与并调控淀粉酶产色链霉菌1628的丰加霉素合成,并为其他生防菌株提高丰加霉素产量提供了良好的理论依据。

关键词: 丰加霉素, 链霉菌, 嘌呤回补途径, 生物农药

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

中图分类号:

S476

张子轩, 宋阳, 申屠旭萍, 俞晓平. 激活嘌呤回补途径提高淀粉酶产色链霉菌丰加霉素产量[J]. 中国生物防治学报, 2023, 39(4): 851-860.

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.

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激活嘌呤回补途径提高淀粉酶产色链霉菌丰加霉素产量

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

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