Please wait a minute...
 
 
食品与发酵工业  2020, Vol. 46 Issue (16): 1-6    DOI: 10.13995/j.cnki.11-1802/ts.023895
  研究报告 本期目录 | 过刊浏览 | 高级检索 |
白色链霉菌ε-聚赖氨酸合酶的异源表达及重组菌全细胞合成ε-聚赖氨酸的条件优化
ITUZE KUBANA Marie Claudine, 乔郅钠, 徐美娟, 陈旭升, 杨套伟, 张显, 邵明龙, 饶志明*
江南大学 生物工程学院, 江苏 无锡,214122
Heterologous expression of Streptomyces albulus ε-poly-L-lysine synthase and optimization of whole-cell biocatalyst for ε-poly-L-lysine synthesis
ITUZE KUBANA Marie Claudine, QIAO Zhina, XU Meijuan, CHEN Xusheng, YANG Taowei, ZHANG Xian, SHAO Minglong, RAO Zhiming*
School of Biotechnology, Jiangnan University, Wuxi 214122, China
下载:  HTML   PDF (3046KB) 
输出:  BibTeX | EndNote (RIS)      
摘要 ε-聚赖氨酸是一种典型的同型L-赖氨酸聚合物,具有广谱抗菌活性和抗噬菌体活性,广泛应用于食品和医药行业。该文克隆了经密码子优化的白色链霉菌Streptomyces albulus来源的ε-聚赖氨酸合酶编码基因pls,首次实现了其在食品安全性菌株枯草芽孢杆菌Bacillus subtilis 168中的异源表达。其次,对B. subtilis 168/pMA5-pls重组菌株生产ε-聚赖氨酸的全细胞催化体系进行了优化,结果表明,在L-赖氨酸质量浓度0.5 g/L, 初始pH和温度分别为3.0和30 ℃条件下,ε-聚赖氨酸合酶的转化效果最好。在最优体系条件下,连续转化4 h,ε-聚赖氨酸的产量达到195.1 mg/L,高于已报道的Bacillus subtilis直接发酵生产的ε-聚赖氨酸产量。该研究构建的全细胞催化体系为食品级ε-聚赖氨酸的生产提供了新策略。
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
ITUZE KUBANA Marie Claudine
乔郅钠
徐美娟
陈旭升
杨套伟
张显
邵明龙
饶志明
关键词:  ε-聚赖氨酸合酶  枯草芽孢杆菌  克隆表达  全细胞催化  ε-聚赖氨酸    
Abstract: ε-poly-L-lysine is a typical L-lysine homopolypeptide with broad-spectrum antibacterial activity and anti-phage activity, which is widely used in food and pharmaceutical industries. Firstly, a codon optimized pls gene, encoding ε-poly-L-lysine synthetase from Streptomyces albulus was cloned, and for the first time realized its heterologous expression in GRAS strain Bacillus subtilis 168. Secondly, the whole-cell catalytic system for the production of ε-poly-L-lysine by the recombinant strain B. subtilis 168/pMA5-pls were optimized. The results showed that at a concentration of 0.5 g/L L-lysine, initial pH and temperature of 3.0 and 30 ℃ respectively, the conversion effect of ε-poly-L-lysine synthetase was best. Finally, under the optimal condition, continuous conversion of 4 hours, yielded approximately 195.1 mg/L of ε-poly-L-lysine, which was higher than the reported yield of ε-poly-L-lysine produced by Bacillus subtilis via direct fermentation. Therefore, the whole-cell catalytic system constructed in this study provides a new strategy for the safe production of ε-poly-L-lysine.
Key words:  ε-poly-L-lysine synthase    Bacillus subtilis    clone expression    whole-cell catalysis    ε-poly-L-lysine
收稿日期:  2020-03-09      修回日期:  2020-03-31      接受日期:  2016-12-20      出版日期:  2020-08-25      发布日期:  2020-09-17      期的出版日期:  2020-08-25
基金资助: 国家重点研发计划项目(2018YFA0900300);国家自然科学基金(31770058);江苏省自然科学基金项目(BK20181205);宁夏回族自治区重点研发计划(2019BCH01002);中组部万人计划科技创新领军人才项目资助
作者简介:  ITUZE KUBANA Marie Claudine(硕士)和乔郅钠(硕士)为共同第一作者(饶志明教授为通讯作者,E-mail:raozhm@jiangnan.edu.cn)
引用本文:    
ITUZE KUBANA Marie Claudine,乔郅钠,徐美娟,等. 白色链霉菌ε-聚赖氨酸合酶的异源表达及重组菌全细胞合成ε-聚赖氨酸的条件优化[J]. 食品与发酵工业, 2020, 46(16): 1-6.
ITUZE KUBANA Marie Claudine,QIAO Zhina,XU Meijuan,et al. Heterologous expression of Streptomyces albulus ε-poly-L-lysine synthase and optimization of whole-cell biocatalyst for ε-poly-L-lysine synthesis[J]. Food and Fermentation Industries, 2020, 46(16): 1-6.
链接本文:  
http://sf1970.cnif.cn/CN/10.13995/j.cnki.11-1802/ts.023895  或          http://sf1970.cnif.cn/CN/Y2020/V46/I16/1
[1] YOSHIDA T,NAGASAWA T.ε-poly-L-lysine: Microbial production, biodegradation and application potential[J].Applied Microbiology and Biotechnology,2003,62(1):21-26.
[2] 石慧,李婵娟,张俊红. ε-聚赖氨酸产生菌及其应用研究概述[J].食品与发酵工业,2016,42(9):263-269.
[3] 张伟娜,李迎秋.ε-聚赖氨酸在食品中应用的进展[J].中国调味品,2012,37(12):5-9.
[4] HYLDGAARD M,MYGIND T,VAD B S,et al.The antimicrobial mechanism of action of ε-poly-L-lysine[J].Applied & Environental Microbiology,2014,80(24):7 758-7 770.
[5] YE Ruosong,XU Hengyi,WAN Cuixiang,et al.Antibacterial activity and mechanism of action of ε-poly-L-lysine[J].Biochemical Biophysical Research Communications,2013,439(1):148-153.
[6] CHEN Xusheng,TANG Lei,LI Shu, et al.Optimization of medium for enhancement of ε-Poly-L-Lysine production by Streptomyces sp. M-Z18 with glycerol as carbon source[J]. Bioresource Technology,2011,102(2):1 727-1 732.
[7] 董难,陈旭升,任喜东,等.发酵过程流加L-谷氨酸提高ε-聚赖氨酸的产量[J].食品与发酵工业,2013,39(7):79-82.
[8] REN Xidong,CHEN Xusheng,ZENG Xin,et al.Acidic pH shock induced overproduction of ε-poly-L-lysine in fed-batch fermentation by Streptomyces sp. M-Z18 from agro-industrial by-products[J].Bioprocess and Biosystems Engineering,2015,38(6):1 113-1 125.
[9] 余明洁,田丰伟,范大明,等.高产ε-聚赖氨酸白色链霉菌的复合诱变选育研究[J].食品工业科技,2008,29(7):99-101;104.
[10] 张超,王正刚,段作营,等.大剂量紫外诱变选育ε-聚赖氨酸高产菌[J].生物加工过程,2007,5(3):64-68.
[11] 吴光耀,陈旭升,王靓,等.核糖体工程技术选育ε-聚赖氨酸高产菌株[J].微生物学通报,2016,43(12):2 744-2 751.
[12] ZHOU Yongpeng,REN Xidong,WANG Liang,et al.Enhancement of ε-poly-lysine production in ε-poly-lysine-tolerant Streptomyces sp. by genome shuffling[J].Bioprocess & Biosystems Engineering,2015,38(9):1 705-1 713.
[13] SHIH I,SHEN M,VAN Y.Microbial synthesis of poly(ε-lysine) and its various applications[J].Bioresource Technology,2006,97(9):1 148-1 159.
[14] SHI Ci,ZHAO Xingchen,LIU Zonghui,et al.Antimicrobial, antioxidant, and antitumor activity of epsilon ε-poly-L-lysine and citral, alone or in combination[J].Food & Nutrition Research,2016,60(1):31 891.
[15] SINGH M,PATEL S K,KALIA V C.Bacillus subtilis as potential producer for polyhydroxyalkanoates[J].Microbial Cell Factories,2009,8(1): 38.
[16] WESTERS L,WESTERS H,QUAX W J.Bacillus subtilis as cell factory for pharmaceutical proteins: A biotechnological approach to optimize the host organism[J].Biochim et Biophys Acta,2004,1 694(1-3):299-310.
[17] EL-SERSY N A,ABDELWAHAB A E,ABOUELKHIIR S S,et al.Antibacterial and anticancer activity of ε-poly-L-lysine (ε-PL) produced by a marine Bacillus subtilis sp.[J].Journal of Basic Microbiology,2012,52(5):513-522.
[18] WALKER J M.The Protein Protocols Handbook[M].3rd edition T,New Jersey: Humama Press,2009.
[19] KRUGER N J.The bradford method for protein quantitation[J].Methods Mol Biol,1994,32:9-15.
[20] ZENG X,CHEN X S,REN X D,et al.Insights into the role of glucose and glycerol as a mixed carbon source in the improvement of ε-poly-L-lysine productivity[J].Applied Biochemistry Biotechnology,2014,173(8):2 211-2 224.
[21] WANG L,LI S,ZHAO J,et al.Efficiently activated ε-poly-L-lysine production by multiple antibiotic-resistance mutations and acidic pH shock optimization in Streptomyces albulus[J].Microbiologyopen,2019,8(5):e00 728.
[22] YAMANAKA K,MARUYAMA C,TAKAGI H,et al.ε-poly-L-lysine dispersity is controlled by a highly unusual nonribosomal peptide synthetase[J].Nature Chemical Biology,2008,4(12):766-772.
[23] SHEN W C,YANG D,RYSER J P.Colorimetric determination of microgram quantities of polylysine by trypan blue precipitation[J].Analytical Biochemistry,1984,142(2):521-524.
[24] MAEDA S,KUNIMOTO K K,SASAKI C,et al.Characterization of microbial poly (ε-L-lysine) by FT-IR, Raman and solid state 13C NMR spectroscopies[J].Journal of Molecular Structure,2003,655(1):149-155.
[25] 黄静敏,吴清平,刘盛荣,等.ε-聚赖氨酸产生菌新菌株的筛选和产物结构鉴定[J].微生物学通报,2011,38(6):871-877.
[26] NISHIKAWA M.Molecular mass control using polyanionic cyclodextrin derivatives for the ε-poly-L-lysine biosynthesis by Streptomyces[J].Enzyme and Microbial Technology,2009,45(4):295-298.
[27] GUSTAFSSON C,GOVINDARAJAN S,MINSHULL J.Codon bias and heterologous protein expression[J].Trends in Biotechnology,2004,22(7):346-353.
[28] SHIMA S,MATSUOKA H,IWAMOTO T,et al.Antimicrobial action of ε-poly-L-lysine[J]. Journal of Antibiotics,1984,37(11):1 449-1 455.
[29] SHIMA S,OSHIMA S,SAKAI H.Biosynthesis of ε-poly-L-lysine by washed mycelium of Streptomyces albulus no. 346[J].Journal of the Agricultural Chemical Society of Japan,1983,57(3):221-226.
[1] 唐璎, 邓展瑞, 黄佳, 杨晓楠. 黄曲霉毒素B1降解菌株的鉴定及降解产物研究[J]. 食品与发酵工业, 2021, 47(7): 64-70.
[2] 钱蕾, 刘延峰, 李江华, 刘龙, 堵国成. 适应性进化和改造质粒稳定性促进枯草芽孢杆菌合成N-乙酰神经氨酸[J]. 食品与发酵工业, 2021, 47(5): 1-6.
[3] 杨心萍, 宋词, 张伟豪, 刘艳, 王洲, 薛正莲. 常压室温等离子体与5-溴尿嘧啶复合诱变及快速选育腺苷高产菌株[J]. 食品与发酵工业, 2020, 46(9): 73-77.
[4] 张大伟, 刘德华, 黄钦钦, 田亚平. 食品级高产亮氨酸氨肽酶重组Bacillus subtilis的构建和发酵优化[J]. 食品与发酵工业, 2020, 46(8): 1-6.
[5] 胡凡, 宿玲恰, 吴敬. Thermobifida fusca麦芽三糖淀粉酶的重组表达及其在麦芽三糖制备中的应用[J]. 食品与发酵工业, 2020, 46(5): 23-30.
[6] 郭佳欣, 张培基, 刘丁玉, 洪坤强, 陈涛, 王智文. 常压室温等离子体诱变选育高产核黄素枯草芽孢杆菌[J]. 食品与发酵工业, 2020, 46(4): 28-33.
[7] 付云, 赵谋明, 卢美杉, 余炼, 刘小玲. 枯草芽孢杆菌YA215发酵螺旋藻渣产抑菌活性的工艺[J]. 食品与发酵工业, 2020, 46(4): 146-152.
[8] 王换男, 卿琳华, 童群义. ε-聚赖氨酸对鲜榨血橙汁贮藏品质的影响[J]. 食品与发酵工业, 2020, 46(4): 219-226.
[9] 郑亚伦, 夏瑛, 李良, 董孝元, 方尚玲, 陈茂彬, 李琴. 源于解淀粉芽孢杆菌酸性木聚糖酶酶学性质的研究[J]. 食品与发酵工业, 2020, 46(24): 58-65.
[10] 李秋莹, 张婧阳, 孙彤, 谢晶, 邓尚贵, 林洪, 郭晓华, 励建荣. ε-聚赖氨酸及其复合保鲜技术在水产品保鲜中的研究进展[J]. 食品与发酵工业, 2020, 46(22): 263-269.
[11] 张丽杰, 张怀志, 徐岩. 枯草芽孢杆菌Nr.5和底物添加促进酱油中吡嗪类物质合成[J]. 食品与发酵工业, 2020, 46(21): 1-8.
[12] 王梓源, 李欣颖, 吕俊阁, 付萍, 孙雪文, 李雪晶, 谭之磊, 贾士儒. ε-聚赖氨酸对大肠杆菌的抑菌机制[J]. 食品与发酵工业, 2020, 46(21): 34-41.
[13] 陈彬和, 赵炳天, 孙亚娟, 李云兴. 大豆发酵液的抗氧化活性[J]. 食品与发酵工业, 2020, 46(17): 119-124.
[14] 冒鑫哲, 彭政, 周冠宇, 堵国成, 张娟. 枯草芽孢杆菌高产角蛋白酶发酵条件优化[J]. 食品与发酵工业, 2020, 46(17): 138-144.
[15] 卢超, 陈景鲜, 王国霞, 陈刚, 李春阁, 王会鱼. 枯草芽孢杆菌L07产中性蛋白酶发酵条件优化[J]. 食品与发酵工业, 2020, 46(16): 148-153.
No Suggested Reading articles found!
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
版权所有 © 《食品与发酵工业》编辑部
地址:北京朝阳区酒仙桥中路24号院6号楼111室
本系统由北京玛格泰克科技发展有限公司设计开发  技术支持:support@magtech.com.cn