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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2018, Vol. 38 Issue (10): 20-29    DOI: 10.13523/j.cb.20181003
Orginal Article     
Isolation and Identification of Antibacterial Lipopeptides Fengycin Produced by Bacillus amyloliquefaciens TF28 and Its Anti-fungal Mechanism Studies
Yu-shuai LIU1,2,Jie ZHANG3,Jin ZHONG3,Jing LI1,2,Li-qiang MENG1,2,Shu-mei ZHANG1,2,**()
1 Institute of Microbiology of Heilongjiang Academy of Sciences, Harbin 150010, China
2 Institute of Advanced Technology, Heilongjiang Academy of Sciences, Harbin 150020, China
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Abstract  

The antifungal lipopeptides produced by Bacillus amyloliquefaciens TF28 have been isolated and identified and their antifungal activity have been studied. The extract of antifungal lipopeptides was prepared by acid precipitation, ethyl acetate and methanol extraction. After two rounds of HPLC separation, eight antifungal lipopeptides were obtained within 32~42 min of retention time, and they have been identified as fengycins by MALDI-TOF-MS, which show strong antifungal activity against Fusarium oxysporum and Fusarium graminearum. These results will contribute to the directional genetic manipulation of the strain TF28 to improve the yield of antifungal lipopeptides.

Key words： Bacillus amyloliquefaciens      Fusarium oxysporum      Fusarium graminearum      Antifungal lipopeptides      Fengycin     
Received: 14 May 2018      Published: 09 November 2018
ZTFLH:  Q939  
Corresponding Authors: Shu-mei ZHANG     E-mail: 1401135157@qq.com
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Yu-shuai LIU
Jie ZHANG
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Jing LI
Li-qiang MENG
Shu-mei ZHANG
Cite this article:

Yu-shuai LIU,Jie ZHANG,Jin ZHONG,Jing LI,Li-qiang MENG,Shu-mei ZHANG. Isolation and Identification of Antibacterial Lipopeptides Fengycin Produced by Bacillus amyloliquefaciens TF28 and Its Anti-fungal Mechanism Studies. China Biotechnology, 2018, 38(10): 20-29.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20181003     OR     https://manu60.magtech.com.cn/biotech/Y2018/V38/I10/20

Fig.1 Antifungal activity of lipopeptide extract produced by strain TF28 to pathogenic fungi 1、3:Lipopeptide extract;2、4:Control (a) The antifungal activity to Fusarium oxysporum (b) The antifungal activity to Fusarium graminearum
Fig. 2 Analysis of the lipopeptide extract produced by strain TF28 by HPLC 1-11:constituents 1-11
Fig.3 Antifungal activity of first HPLC separation constituents to Fusarium oxysporum 1-5、1-6、1-7:Antibacterial active constituents;1-12:Control
Fig.4 Analysis of the second HPLC separation of three active constituents (a) Constituent 1-5 (b) Constituent 1-6 (c) Constituent 1-7 (d) Magnification of the main peaks for constituent 1-7
Fig.5 Antifungal activity of each constituent from the second HPLC separation (a) The antifungal activity to Fusarium oxysporum (b)~(d) The antifungal activity to Fusarium graminearum 2-1~2-8: Each active constituent ; 2-9~2-13: Control
序号
Constituents		 相对分子量
[M +H]+		 特征离子峰1
Characteristic ion peak 1		 特征离子峰2
Characteristic ion peak 2
2-1 1463.7832 966 1 080
2-2 1491.8247 994 1 108
2-3 1505.8154 994 1 108
2-4 1447.7803 966 1 080
2-5 1491.7981 966 1 080
2-6 1475.8151 994 1 108
2-7 1519.8358 994 1 108
2-8 1489.8183 994 1 108
Table 1 Results of MALDI-TOF spectrum of 8 active constituents
Fig.6 MALDI-TOF spectrum of 8 active constituents of strain TF28 (a)-(h): [M +H]+ of constituents 2-1~2-8
Fig.7 MALDI-TOF /TOF spectrum of procured ions of 8 active constituents of strain TF28 (i)~(p): Characteristic ion peaks of constituents 2-1~2-8
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