食品科学

• 生物工程 • 上一篇    下一篇

1株产脂肽类抗生素芽孢杆菌的鉴定及脂肽类抗生素相关基因克隆

冉军舰1,2,徐剑宏1,胡晓丹1,史建荣1,*   

  1. 1.江苏省农业科学院食品质量安全与检测研究所,江苏省食品质量安全重点实验室-省部共建国家重点实验室培育基地,农业部农产品质量安全控制技术与标准重点实验室,江苏省转基因安全评价公共服务中心,江苏省现代粮食流通与安全协同创新中心,江苏 南京 210014;2.河南科技学院食品学院,河南 新乡 453003
  • 出版日期:2016-09-15 发布日期:2016-09-22

Identification of a Bacillus Strain Producing Lipopeptide and Cloning of Genes Related to Lipopeptide

RAN Junjian1,2, XU Jianhong1, HU Xiaodan1, SHI Jianrong1,*   

  1. 1. Key Laboratory of Food Quality and Safety of Jiangsu Province-State Key Laboratory Breeding Base, Key Laboratory of Control
    Technology and Standard for Agro-product Safety and Quality, Ministry of Agriculture, Jiangsu Center for Genetically Modified
    Organisms Evaluation and Detection, Collaborative Innovation Center for Modern Grain Circulation and Safety, Institute of Food
    Quality Safety and Detection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China; 2. School of Food Science,
    Henan Institute of Science and Technology, Xinxiang 453003, China
  • Online:2016-09-15 Published:2016-09-22

摘要:

从小麦根际土壤中分离得到一株对禾谷镰刀菌有拮抗作用的菌株7F1。以菌株7F1基因组总DNA为模板,通过16S rDNA和gyrB基因序列分析,鉴定7F1为多粘类芽孢杆菌。本实验对该菌种产拮抗物的稳定性进行了研究,同时,应用特异性引物对7F1中脂肽类抗生素合成相关基因进行检测。结果表明:该菌产生的拮抗物在40~90 ℃保持较高的活性;在蛋白酶K、胃蛋白酶和胰蛋白酶处理后,抑菌活性显著下降;在pH 3.0~10.0具有抑菌活性;使用已报道的脂肽类抗生素基因片段设计的引物进行聚合酶链式反应扩增,从该菌的基因组DNA中扩增得到了3 种bamC、eriSa、ituC脂肽类抗生素合成酶基因。此研究结果对脂肽类抗生素进一步的分离提供了重要参考。

关键词: 脂肽类抗生素, 芽孢杆菌, 鉴定, 抑菌活性, 基因克隆

Abstract:

A strain named 7F1 which showed strong inhibitory effect on Fusarium graminearum was isolated from
rhizosphere soil of healthy wheat. Using its genomic DNA as the polymerase chain reaction (PCR) template, strain 7F1 was
identified as Paenibacillus polymyxa by 16S rDNA and gyrB sequence analysis. The stability of its antagonistic activity
was evaluated and the detection of lipopeptide synthesis-related genes was conducted by using specific primers. The results
showed that the antagonist produced by strain 7F1 had good heat stability, which could remain stable at 40–90 ℃. The
inhibitory activity was markedly decreased by treatment with proteinase K, pepsin or trysin. The antagonist could exert its
activity in the pH range from 3.0 to 10.0. Using previously reported lipopeptide primers, the lipopeptide synthase genes
bamC, eriSa and ituC were successfully amplified by PCR from the genomic DNA of strain 7F1. These results may provide
an important reference for further separation and application of lipopeptide.

Key words: lipopeptide, Bacillus, identification, antimicrobial activity, gene cloning

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