抗菌豆豉发酵菌株的筛选及其脂肽组分鉴定和特性研究

中国生物工程杂志

2013, Vol. 33

Issue (7): 50-56

研究报告

抗菌豆豉发酵菌株的筛选及其脂肽组分鉴定和特性研究

孙力军, 王雅玲, 刘唤明, 徐德峰, 张永平, 聂芳红

广东省水产品加工与安全重点实验室广东海洋大学食品科技学院湛江 524088

Identification of Antimicrobial Lipopetides Component Produced by Isolate from Douchi and Its Antimicrobial Properties

SUN Li-jun, WANG Ya-ling, LIU Huan-min, XU De-feng, ZHANG Yong-pin, NIE Fang-hong

Guangdong Provincial Key Laboratory of Aquatic Products Processing and Safety, College of Food Science Technology, Guangdong Ocean University, Zhanjiang 524088, China

全文: PDF(1574 KB) HTML

摘要： 目的:从豆豉中分离广谱抗菌活性菌株,并鉴定其发酵产物中的抗菌成分。方法:采用生理生化实验结合16SrRNA序列测定法鉴定目标分离株,通过LC-MS和ESI/CID技术对其抗菌组分进行鉴定。结果:从8个豆豉样品中分离出7株具有抗菌活性菌株,其中抗菌活性较强的分离株NT-6为枯草芽孢杆菌(Bacillus subtilis),其发酵液抗菌提取物的主要成分是抗菌脂肽类物质iturin、fengycin、surfactin同系物的混合物。该提取物对供试革兰氏阳性、阴性细菌、霉菌等指示菌具有广谱的抗菌活性,在pH 2~12、121℃条件下加热30min活性不丧失,说明具有广泛的pH适应范围和良好的耐热性。结论:该菌株产生多组分抗菌脂肽,其抑菌谱宽,适应性好,显示了在食品、农业、医药等领域具有良好的应用前景。

关键词： 豆豉; 纳豆菌; 抗菌活性; 抗菌脂肽

Abstract: Objective: The paper report that screening broad-spectrum antimicrobial activity strain from douchi and identification of antimicrobial component of fermented extracts produced by the isolate. Methods: Through physiological and biochemical experiments and 16SrRNA sequencing analysis, the isolate identification was carried out. Analysis of the related antimicrobial substances was carried out by high performance liquid chromatography (HPLC)-electrospray ionization (ESI)-mass spectrometer (MS). Results: Seven antimicrobial activity isolates were isolated from eight douchi samples. The isolate, strain NT-6 with stronger antimicrobial activity, was identified as Bacillus subtilus, which produced three families of antimicrobial lipopeptides, iturins and fengycins and surfactins. Culture filtrate extracts of strain NT-6 strongly inhibited the growth of all Gram-positive bacteria tested except Bacillus subtilus and all Gram-negative ones and all fungi except Fusarium oxysporum f. sp. vasinfectum. And, antimicrobial activity of the extracts was stable under pH 2~12 and 121℃ 30min conditions. Conclusion: Therefore, it shows good application prospects in the food, agriculture, medicine and other fields that the isolate can produce more antimicrobial lipopeptide ingredients, a wide antimicrobial spectrum, and good adaptability.

Key words: Douchi Bacillus subtilus Antimicrobial activity Antimicrobial lipopeptide

收稿日期: 2012-08-08 出版日期: 2013-07-25

ZTFLH:

Q815

基金资助: 国家自然科学基金(30972287);"十二五"国家支撑计划项目(2012BAD29B06);广东省教育部产学研结合项目(2011B090400154);湛江科技计划(2011D02)资助项目

通讯作者: 王雅玲 E-mail: wangylchina@163.com

	服务
	把本文推荐给朋友
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章
	孙力军
	王雅玲
	刘唤明
	徐德峰
	张永平
	聂芳红

引用本文:

孙力军, 王雅玲, 刘唤明, 徐德峰, 张永平, 聂芳红. 抗菌豆豉发酵菌株的筛选及其脂肽组分鉴定和特性研究[J]. 中国生物工程杂志, 2013, 33(7): 50-56.

SUN Li-jun, WANG Ya-ling, LIU Huan-min, XU De-feng, ZHANG Yong-pin, NIE Fang-hong. Identification of Antimicrobial Lipopetides Component Produced by Isolate from Douchi and Its Antimicrobial Properties. China Biotechnology, 2013, 33(7): 50-56.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2013/V33/I7/50

[1] 高瑞萍,刘辉,刘嘉,等. 豆豉的研究进展. 食品与发酵科技,2011,47(1):23-26. Gao R P, Liu H, Liu J, et al. The research developments on douchi. Food and Fermentation Technology, 2011,47(1):23-26.
[2] 黄占旺,魏萍,刘海林,等. 纳豆菌生物学特性研究.江西农业大学学报,2004,26(1):83-85. Huang Z W, Wei P, Liu H L, et al. A study on biological properties of Bacillus natto. Acta Agriculturae Universitatis Jiangxiensis, 2004, 26(1):83-85.
[3] 蒋立文, 周传云,黄香华. 纳豆菌的研究现状和应用进展. 中国食物与营养,2007, 6:24-26. Jiang L W, Zhou C Y, Huang X H. The current research status and application development of Bacillus natto. Chinese Food and Nutrion,2007,6:24-26.
[4] 纪宁,孔繁东,祖国仁,等. 纳豆菌抗菌作用的研究现状及展望. 食品研究与开发, 2006, 27 (1): 138-142. Ji N, Kong F D, Zu G R, et al. The present situation and developmental tendency of antimicrobial functions of bacillus natto. Food Research and Development,2006,27(1):138-142.
[5] 钟青萍,谢俊杰,余世望. 纳豆菌的分离鉴定及其抗菌活性. 食品科学,2002,23(10):109-112. Zong Q P, Xie J J, Yu S W. Isolation,identification and antimicrobial activity assay of Bacillus natto. Food Science,2002,23(10):109-112.
[6] Cao X H, Liao Z Y, Wang C L, et al. Evaluation of a lipopeptide biosurfactant from Bacillus natto TK-1 as a potential. Source of anti-adhesive, antimicrobial and antitumor activities. Brazilian Journal of Microbiology, 2009, 40: 373-379.
[7] Sumi H. Antibacterial activity of Bacillus natto growth inhibition against Escherichia coli O157. Bioindustry, 1997, 14:47.
[8] Ozawa K, Yagu U, Yamanaka K. Antagonistic effects of Bacillus natto and Streptococcus faecalis on growth of Candida albicans. Journal of Microbiology Immunoogy, 1979, 23(12): 1147-1156.
[9] Sun L J, Lu Z X, Bie X M, et al. Isolation and characterization of a co-producer of fengycins and surfactins, endophytic Bacillus amyloliquefaciens ES-2, from Scutellaria baicalensis Georgi. World Journal of Microbiology Biotechnology, 2006, 12: 1259-1266.
[10] Weisburg W G, Barns S M, Pelletier D A, et al. 16S ribosomal DNA amplification for phylogenetic study. Journal of Bacteriology, 1991, 173: 697-703.
[11] 孙力军.植物内生菌Bacillus amyloliquefaciens ES-2的分离筛选及其抗菌物质的研究.南京:南京农业大学,2006. Sun L J. Isolation of endophytic Bacillus amyloloquefaciens ES-2 and Studies on Antimicrobial Substances. Nanjing: Nanjing Agricutural University,2006.
[12] Kim P I, Bai H, Bai D, et al. Purification and characterization of a lipopeptide produced by Bacillus thuringiensis CMB26. Journal of Applied Microbiology, 2004, 97: 942-949S.
[13] Koumoutsi A, Chen X H, Henne A, et al. Structural and functional characterization of gene clusters directing nonribosomal synthesis of bioactive cyclic lipopeptides in Bacillus amyloliquefaciens strain FZB42. Journal of Bacteriology, 2004, 186: 1084-1096.
[14] Kowall M, Vater J, Kluge B, et al. Separation and characterization of surfactin isoforms produced by Bacillus subtilis OKB 105. Journal of Colloid Interface Science, 1998, 204: 1-8.
[15] 陈华,王丽,袁成凌,等.高效液相色谱-电喷雾质谱法分离和鉴别枯草芽孢杆菌产生的脂肽类化合物. 色谱, 2008, 26(3): 343-347. Chen H, Wang L, Zhen ZH M,et al. Isolation and identification of lipopeptides produced by Bacillus subtilis using high performance liquid chromatography and electrospray ionization mass spectrometry. Chinese Journal of Chromatography, 2008,26(3):343-347.
[16] Vater J, Kablitz B, Wilde C, et al. Matrix-assisted laser desorption ionization-time of flight mass spectrometry of lipopeptide biosurin whole cells and culture filtrates of Bacillus subtilis C-1 isolated from petroleum sludge. Applied and Environmental Microbiology, 2002, 68: 6210-6219.
[17] Roongawang N, Thaniyavarn J, Thaniyavarn S, et al. Isolation and characterization of a halotolerant Bacillus subtilis BBK-1 which produces three kinds of lipopeptides: bacillomycin L, plipastatin, and surfactin. Extremophiles, 2002, 6: 499-506.
[18] Tang J S, Zhao F, Gao H, et al. Characterization and online detection of surfactin isomers based on HPLC-MSⁿ analyses and their inhibitory effects on the overproduction of nitric oxide and the release of TNF-α and IL-6 in LPS-induced macrophages. Marine Drugs, 2010, 8(10): 2605-2618.

[1]	戈家傲,刘畅,龚建刚,刘艳琴. 抗菌环肽的研究进展 ^*[J]. 中国生物工程杂志, 2018, 38(11): 76-83.
[2]	刘宇帅,张杰,钟瑾,李晶,孟利强,张淑梅. 解淀粉芽孢杆菌TF28抗菌脂肽芬芥素的分离鉴定及抑菌作用 ^*[J]. 中国生物工程杂志, 2018, 38(10): 20-29.
[3]	巫春旭, 卢雪梅, 金小宝, 朱家勇. 天蚕素类抗菌肽分子设计研究进展[J]. 中国生物工程杂志, 2016, 36(2): 96-100.
[4]	陶思美, 郑维, 赵朋超, 周伟, 权春善, 范圣第. bmy基因敲除对解淀粉芽孢杆菌Q-426溶血性及抗菌活性的影响[J]. 中国生物工程杂志, 2014, 34(3): 56-60.
[5]	李建波, 江明锋, 王永. 藏绵羊乳腺溶菌酶基因的克隆、原核表达及抗菌活性研究[J]. 中国生物工程杂志, 2013, 33(8): 38-44.
[6]	明飞平, 杨军, 朱进美, 邝哲师, 李华周, 夏枫耿, 叶明强, 王候光, 赵祥杰, 黄志丰, 蔡海明, 施巨清, 马苗鹏, 张玲华. 5’非翻译区序列改建提高抗菌肽PR39表达[J]. 中国生物工程杂志, 2013, 33(12): 86-91.
[7]	熊文, 杨学敏, 王建华, 权春善, 范圣第. DKPs对解淀粉芽孢杆菌Q-426抗菌活性物质基因表达量的影响[J]. 中国生物工程杂志, 2012, 32(03): 47-52.
[8]	魏浩, 陆兆新, 吕凤霞, 翟亚楠, 郭昊, 郝慧, 别小妹. Bacillus amyloliquefaciens ES-2发酵产抗菌脂肽消泡剂的筛选及脂肽的提取和纯化[J]. 中国生物工程杂志, 2011, 31(02): 85-90.
[9]	曹小红,廖振宇,王春玲,哈志瑞,杨亚静,鲁梅芳,励建荣. Bacillus natto TK-1产脂肽的纯化、抑菌活性及其表面活性剂特性[J]. 中国生物工程杂志, 2008, 28(1): 44-48.
[10]	徐灵龙王云峰石星明童光志. 抗菌肽及其功能研究[J]. 中国生物工程杂志, 2007, 27(1): 115-118.
[11]	高必达. 丁香假单胞菌环式脂肽毒素的生理和分子生物学研究[J]. 中国生物工程杂志, 2000, 20(4): 55-59.
[12]	郭玉梅, 戴祝英. 昆虫抗菌肽的研究进展[J]. 中国生物工程杂志, 1996, 16(3): 24-27.
[13]	张震元. 具有类似β内酰胺作用的新型母核的新抗生素Lactivicin[J]. 中国生物工程杂志, 1987, 7(6): 65-65.

Viewed

Full text

Abstract

Cited

Shared

Discussed