基于NRPSs基因筛选与鉴定海芦笋内生细菌

详细信息    查看全文 | 推荐本文 |

英文篇名：Screening and identification of endophytic bacteria from Salicornia bigelovii. Torr based on a NRPS functional gene assay 作者：李代婧 ; 殷跃 ; 刘梦婕 ; 卫婷 ; 郭佳 ; 辛志宏 英文作者：LI Dai-jing;YIN Yue;LIU Meng-jie;WEI Ting;GUO Jia;XIN Zhi-hong;College of Food Science and Technology,Nanjing Agricultural University; 关键词：内生细菌 ; 分离鉴定 ; 系统发育分析 ; 非核糖体多肽合成酶 ; 质谱 ; 环肽 英文关键词：endophytic bacillus;;isolation and identification;;phylogenetic analysis;;nonribosomal peptide synthetase;;mass spectrum;;cyclopeptide 中文刊名：SPFX 英文刊名：Food and Fermentation Industries 机构：南京农业大学食品科技学院; 出版日期：2017-07-20 10:10 出版单位：食品与发酵工业 年：2017 期：v.43;No.357 基金：2017年农产品质量安全风险评估项目(GJFP201701102);; 2016年南京农业大学大学生创新训练计划(SRT)项目(1618C01) 语种：中文; 页：SPFX201709010 页数：6 CN：09 ISSN：11-1802/TS 分类号：62-67

摘要

利用形态学观察和分子生物学相结合的方法分离鉴定海芦笋内生细菌,并筛选含有NRPS基因的菌株(nonribosomal peptide synthetase,NRPS),以期获得能够产生环肽化合物的菌株。研究结果表明,共从海芦笋中分离到6株内生细菌Srtli-F1~Srtli-F6,分别鉴定为绿针假单胞菌橙黄亚种(Pseudomonas chlororaphis subsp.aurantiaca)、组氨酸节杆菌(Arthrobacter histidinolovorans)、庆盛芽孢杆菌(Bacillus qingshengii)、类芽孢杆菌羽扇豆根瘤菌(Paenibacillus lupini)、假蕈状芽孢杆菌(Bacillus pseudomycoides)、高山麝病原性阿尔伯蒂埃希氏杆菌(Escherichia albertii),其中菌株Srtli-F5检测到含有NRPS基因,经系统发育树分析,预测该菌株可能产生环肽类化合物,该株菌经过发酵,有机溶剂萃取发酵产物,并用电喷雾质谱ESI-MS(electrospray mass,ESI-MS)分析Srtli-F5能产生环肽化合物——杆菌肽,证实了该预测结果。该研究为定向筛选能够产生环肽类化合物的菌种开拓了新思路。
        In the current research,endophytic bacteria were isolated from Salicornia bigelovii. Torr and characterized by morphological observation and molecular methods,and the NRPS gene was screened to obtain the strains that could produce cyclic peptide compounds. The results showed that six endophytic bacteria were isolated and identified as Pseudomonas chlororaphis subsp. Aurantiaca,Arthrobacter histidinolovorans,Bacillus qingshengii,Paenibacillus lupini,Bacillus pseudomycoides,Escherichia albertii,respectively. Among them,the strain Srtli-F5 contained NRPS genes and was predicted to have capability of producing cyclopeptide compounds by phylogenetic tree analysis. It was further confirmed by the detection of fermentation production using ESI-MS( electrospray mass,ESI-MS) that the strain really produced bacitracin. The study provides a new method for directional screening strains with the potential to produce cyclic peptide compounds.

引文

[1]冉火苗,孔望君,蒋会芳,等.盐生海芦笋抗菌内生细菌的筛选与鉴定[J].食品与发酵工业,2016,42(3):79-86.
    [2]陆东和,张慜,蔡金龙,等.海芦笋营养价值及其采后品质变化[J].食品工业科技,2010,31(6):303-305.
    [3]朱鹏.珍珠膜海绵共附生微生物PKS与NRPS的筛选与模块结构研究[D].杭州:浙江大学,2008.
    [4]MOORE B S,Biosynthesis of marine natural products:microorganisms(Part A)[J].NATURAL PRODUCT REPORTS,2005,22(5):580-593.
    [5]文艳苹.基于PKS、NRPS基因的抗生素paenimacrolidin和嗜铁素paenibactin研究[D].杭州:浙江大学,2011.
    [6]LANGE A,SUN H,PILGERJ,et al.Predicting the structure of cyclic lipopeptides by Bioinformatics:structure revision of Arthrofactin[J].CHEMBIOCHEM,2012,13(18):2 671-2 675.
    [7]刘怡君,潘沐,王明洋,等.海芦笋内生真菌基于PKSⅠ型功能基因的分离与鉴定[J].食品科学,2014(11):115-119.
    [8]K YAMANAKA,KA REYNOLDS,RD KERSTEN,et al.Direct cloning and refactoring of a silent lipopeptide biosynthetic gene cluster yields the antibiotic taromycin A[J].Proceedings of the National Academy of Sciences,2014,111(5):1 957-1 962.
    [9]周培瑾,徐毅,马允卿,等.极端嗜盐菌16S r DNA的PCR扩增[J].微生物学报,1994,24(1):6-8.
    [10]胡尚勤.杆菌肽高产的代谢调控[J].国外医药(抗生素分册),2004(4):160-162.
    [11]周岩民,王龙昌.杆菌肽产品在动物饲料中的应用研究进展[J].饲料工业,2012(18):1-5.
    [12]陈吉红,庞玉红,郭荣富.杆菌肽锌在畜禽中的应用及其安全性评价[J].中国饲料,2004(9):28-29.
    [13]RAVINDRAN V,THOMAS D V,THOMAS D G,et al.Performance and welfare of broilers as affected by stocking density and zinc bacitracin supplementation[J].Animal Science Journal,2006,77(1):110-116.
    [14]周臣飞,李俊辉,季朝金.杆菌肽类饲料添加剂在畜禽养殖中的应用[J].中国动物保健,2011(12):8-11.
    [15]周欣,李宝库.杆菌肽的研究进展及应用[J].科技信息,2010(14):92-93.