基于PKS和NRPS基因的海洋来源抗病原菌活性菌株筛选及其代谢产物活性分析  

Screening of marine resistant strain based on PKS and NRPS genes and the activity of its metabolites

作　　者：张丽影 刘骏锋 董极靓 金黎明 许永斌 郑维 权春善 ZHANG Liying;LIU Junfeng;DONG Jiliang;JIN Liming;XU Yongbin;ZHENG Wei;QUAN Chunshan(Key Laboratory of Biotechnology and Bioresources Utilization(Ministry of Education),College of Life Science,Dalian Minzu University,Dalian 116600,Liaoning,China)

机构地区：[1]大连民族大学生命科学学院、生物技术与资源利用教育部重点实验室,辽宁大连116600

出　　处：《生物工程学报》2022年第12期4520-4535,共16页Chinese Journal of Biotechnology

基　　金：辽宁省教育厅重点实验室项目(LS2010049)。

摘　　要：基于聚酮合成酶基因(polyketide synthases gene,PKS)和非核糖体多肽合成酶基因(non ribosomal polypeptide synthase gene,NRPS),本研究从77株分离于北冰洋海泥的菌株中筛选出1株具有较高抗病原菌活性的菌株并对其进行了菌种鉴定。通过优化培养基组成和发酵条件提高了该菌株活性代谢产物的产量,并利用高分辨率质谱(high resolution mass spectrometry,HRMS)、核磁氢谱(1H nuclear magnetic hydrogen,1H NMR)和碳谱(13C NMR)对其主要代谢产物进行了结构鉴定。测定了该菌株主要代谢产物的抗菌谱及代谢产物对黄瓜枯萎病的影响。研究结果表明,该菌株为贝莱斯芽孢杆菌(Bacillus velezensis),其对植物具有一定的促生作用。当发酵条件为麦芽糖5 g/L、胰蛋白胨10 g/L、氯化钠10 g/L、温度30℃、转速150 r/min、发酵时间60 h时,该菌株代谢产物的抑菌圈直径由(16.23±0.42)mm提高至(24.42±0.57)mm。菌株代谢产物含有大环内酯类化合物macrolactin A,其对多种病原细菌和真菌具有明显拮抗作用。黄瓜幼苗实验表明,该菌株代谢产物对黄瓜枯萎病具有防护作用,其作为生防菌剂具有一定的开发应用潜力。Based on polyketide syntheses gene(PKS)and non-ribosomal peptide synthetases gene(NRPS),one strain with high anti-pathogenic activity was screened from 77 strains isolated from Arctic marine sediments and identified.By optimizing the composition of culture medium and fermentation conditions,the production of this strain’s active metabolites was improved and the main metabolites were identified by HRMS,1H NMR and 13C NMR.The antibacterial spectrum of the main metabolites and the effect of the metabolites on cucumber Fusarium wilt were also determined.The results showed that the strain was Bacillus velezensis and it showed growth promoting effect on plants.When the strain was cultured in 5 g/L maltose,10 g/L tryptone,10 g/L sodium chloride,at 30℃,150 r/min for 60 h,the diameter of the inhibition zone increased from(16.23±0.42)to(24.42±0.57)mm.The metabolites of this strain mainly contain macrolide compound macrolactin A,which has antagonistic effect on a variety of pathogenic bacteria and fungi.Cucumber seedling experiments showed that the metabolites of this strain had a protective effect on cucumber Fusarium wilt,and showed a good potential for development and application as a biocontrol agent.

关 键 词：海洋拮抗菌 贝莱斯芽孢杆菌 代谢产物 黄瓜枯萎病 

分 类 号：TQ450.1[化学工程—农药化工]