Please wait a minute...

中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2016, Vol. 36 Issue (7): 55-63    DOI: 10.13523/j.cb.20160709
技术与方法     
炭样小单孢菌中抗生素生物合成相关蛋白的筛选
陈娟, 杨慧林, 黄运红, 龙中儿
江西师范大学生命科学学院 南昌 330022
Screening of the Protein Related to Antibiotics Biosynthesis in Micromonospora carbonacea JXNU-1
CHEN Juan, YANG Hui-lin, HUANG Yun-hong, LONG Zhong-er
College of Life Sciences, Jiangxi Normal University, Nanchang 330022, China
 全文: PDF(1479 KB)   HTML
摘要:

目的:筛选一株具有广谱抗菌活性的炭样小单孢菌JXNU-1中核苷类抗生素生物合成相关蛋白。方法:通过iTRAQ定量蛋白质组学技术对JXNU-1菌体生长期(36h)和产物合成期(108h)的差异蛋白进行鉴定和功能分析。结果:基于iTRAQ定量蛋白质组学技术共鉴定出炭样小单孢菌总蛋白质2390个,差异表达蛋白172个,在产物合成期(108h)表达上调76个、表达下调96个。通过蛋白GO和COG注释等功能分析,筛选出12个与抗生素合成密切相关蛋白和5个生物合成基因簇。结论:利用iTRAQ技术筛选出炭样小单孢菌JXNU-1的抗生素合成相关蛋白,为阐明该抗生素的生物合成机制奠定实验依据。

关键词: 蛋白质组学生物合成途径基因簇炭样小单孢菌抗生素    
Abstract:

Objective:To screen the protein related to antibiotics biosynthesis in Micromonospora carbonacea JXNU-1 with broad-spectrum antimicrobial activity. Methods:iTRAQ was used to analyze the differentially expressed proteins between the time of antibiotics secretion/non-secretion in M.carbonacea JXNU-1. Results:A total of 2 390 proteins were identified using iTRAQ, and 172 proteins were differentially expressed during the antibiotics secretion time in M.carbonacea JXNU-1, including 76 up-regulated and 96 down-regulated. Twelve proteins and five gene clusters closely related to biosynthesis of antibiotics were screened based on the functional analysis of the differentially expressed proteins with GO and COG, annotation. Conclusion:The proteins related to antibiotics biosynthesis in M.carbonacea JXNU-1 have been screened based on iTRAQ, it could provide the theoretical basis for revealing the mechanism of antibiotics biosynthesis in M.carbonacea JXNU-1.

Key words: Antibiotic    Micromonospora carbonacea    Biosynthetic pathway    Gene cluster    Proteomics
收稿日期: 2016-01-27 出版日期: 2016-07-25
ZTFLH:  Q591.2  
基金资助:

国家自然科学基金(31160029,31360018),江西省自然科学基金(20132BAB204007,20161BAB204174)资助项目

通讯作者: 龙中儿     E-mail: longzhonger@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  

引用本文:

陈娟, 杨慧林, 黄运红, 龙中儿. 炭样小单孢菌中抗生素生物合成相关蛋白的筛选[J]. 中国生物工程杂志, 2016, 36(7): 55-63.

CHEN Juan, YANG Hui-lin, HUANG Yun-hong, LONG Zhong-er. Screening of the Protein Related to Antibiotics Biosynthesis in Micromonospora carbonacea JXNU-1. China Biotechnology, 2016, 36(7): 55-63.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20160709        https://manu60.magtech.com.cn/biotech/CN/Y2016/V36/I7/55

[1] Chen W,Qi J,Wu P,et al.Natural and engineered biosynthesis of nucleoside antibiotics in Actinomycetes.Journal of Industrial Microbiology&Biotechnology,2015,43(2):1-17.
[2] Niu G,Tan H.Nucleoside antibiotics:biosynthesis,regulation,and biotechnology.Trends in Microbiology,2015,23(2):110-119.
[3] 龙中儿,朱跃进,黄运红,等.一株具有广谱抗菌活性小单孢菌的分离和鉴定.微生物学通报,2008,35(3):378-384.Long Z E,Zhu Y J,Huang Y H,et al.Isolation and identification of a strain of Micromonospora with broad-spectrum antimicrobial activity.Microbiology,2008,35(3):378-383.
[4] 黄运红,高兴强,龙中儿.炭样小单孢菌抗生素抗植物病原真菌作用.农药,2009,48(11):843-845.Huang Y H,Gao X Q,Long Z E.The antibacterial activities of antibiotics produced by Micromonospora carbonacea to plant pathogenic fungi.Agrochemicals,2009,48(11):843-845.
[5] 朱跃进,龙中儿,黄运红,等.一株稀有放线菌发酵产抗生素的工艺研究.化学与生物工程,2006,23(12):39-42.Zhu Y J,Long Z E,Huang Y H,et al.Studies on fermentation technology for a strain of rare Actinomycete producing antibiotics.Chemistry&Bioengineering,2006,23(12):39-42.
[6] 龙中儿,朱跃进,黄运红,等.炭样小单孢菌JXNU-1广谱抗生素产物的分离及其理化性质.微生物学通报,2008,35(9):1450-1454.Long Z E,Zhu Y J,Huang Y H,et al.Isolation and purification of antibiotic from the fermentation broth of Micromonospora carbonacea JXNU-1 and its physical-chemical properties.Microbiology,2008,35(9):1450-1454.
[7] 江云,黄运红,李非,等.一株具有广谱抗菌活性炭样小单孢菌的全基因组序列测定.微生物学通报,2015,42(11):2178-2188.Jiang Y,Huang Y H,Li F,et al.Whole genome sequencing of a Micromonospora carbonacea strain with broad-spectrum antimicrobial activity.Microbiology China,2015,42(11):2178-2188.
[8] Zoghbi M E,Altenberg G A.ATP binding to two sites is necessary for dimerization of nucleotide-binding domains of ABC proteins.Biochemical and Biophysical Research Communications,2014,443(1):97-102.
[9] Li M,Chen Z,Zhang X,et al.Enhancement of avermectin and ivermectin production by overexpression of the maltose ATP-binding cassette transporter in Streptomyces avermitilis.Bioresource Technol,2010,101(23):9228-9235.
[10] Aigle B,Lautru S,Spiteller D,et al.Genome mining of Streptomyces ambofaciens.Journal of Industrial Microbiology&Biotechnology,2014,41(2):251-263.
[11] Vingadassalon A,Lorieux F,Juguet M,et al.Natural combinatorial biosynthesis involving two clusters for the synthesis of three pyrrolamides in Streptomyces netropsis.ACS Chemical Biology,2014,10(2):601-610.
[12] Gloster T M.Advances in understanding glycosyltransferases from a structural perspective.Current Opinion in Structural Biology,2014,28:131-141.
[13] Liu X,Xu Z,Xue S,et al.Characterization of the amicetin biosynthesis gene cluster from Streptomyces vinaceusdrappus NRRL 2363 implicates two alternative strategies for amide bond formation.Applied&Environmental Microbiology,2012,78(7):2393-2401.
[14] Peter F,Guengerich,Andrew W,et al.Unusual cytochrome p450 enzymes and reactions.Journal of Biological Chemistry,2013,288(24):17065-17073.
[15] Dickens M L,Priestley N D,Strohl W R.In vivo and in vitro bioconversion of ε-rhodomycinone glycoside to doxorubicin:functions of DauP,DauK,and DoxA.Journal of Bacteriology,1997,179(8):2641-2650.
[16] Liao G,Li J,Li L,et al.Cloning,reassembling and integration of the entire nikkomycin biosynthetic gene cluster into Streptomyces ansochromogenes lead to an improved nikkomycin production.Microbial Cell Factories,2010,9(4):1-7.
[17] Park S C,Song W S,Yoon S.Structural analysis of a putative SAM-dependent methyltransferase,YtqB,from Bacillus subtilis.Biochemical and Biophysical Research Communications,2014,446(4):921-926.
[18] Tercero J A,Espinosa J C,Lacalle R A,et al.The biosynthetic pathway of the aminonucleoside antibiotic puromycin,as deduced from the molecular analysis of the pur cluster of Streptomyces alboniger.Journal of Biological Chemistry,1996,271(3):1579-1590.
[19] 杜爱芹,吴俊,邓子新,等.杀稻瘟菌素生物合成基因簇的边界确定.微生物学通报,2014,41(7):1318-1325.Du A Q,Wu J,Deng Z X,et al.Determination of the boundary of blasticidin S biosynthetic gene cluster.Microbiology,2014,41(7):1318-1325.
[20] Yin K.Positive correlation between expression level of mitochondrial serine hydroxymethyltransferase and breast cancer grade.OncoTargets and Therapy,2015,8:1069-1074.
[21] Feng J,Wu J,Dai N,et al.Discovery and characterization of BlsE,a radical S-adenosyl-L-methionine decarboxylase involved in the blasticidin S biosynthetic pathway.PloS One,2013,8(7):e68545.
[22] Chen W,Huang T,He X,et al.Characterization of the polyoxin biosynthetic gene cluster from Streptomyces cacaoi and engineered production of polyoxin H.Journal of Biological Chemistry,2009,284(16):10627-10638.
[23] 田云龙.中生菌素生物合成基因簇克隆及分析.北京:中国农业科学院,农业环境与可持续发展研究所研究生院,2010.Tian Y L.Cloning and analysis of Zhongshengmycin Biosynthetic Gene Cluster.Beijing:Chinese Academy of Agricultural Sciences,Graduate School of Agricultural Environment and Sustainable Development,2010.
[24] Lambert D G.Proteomics and metabolomics.Anaesthesia&Intensive Care Medicine,2013,14(4):169-170.
[25] 赵静,王宏伟,田二杰等.蛋白质组学实验技术及其应用.动物医学进展,2015,36(1):116-120.Zhao J,Wang H W,Tian E J,et al.Proteomics experiment technologies and their applications.Progress in Veterinary Medicine,2015,36(1):116-120.
[26] Ma L,Bu D P,Yang Y X,et al.iTRAQ quantitative analysis of plasma proteome changes of cow from pregnancy to lactation.Journal of Integrative Agriculture,2015,14(7):1407-1413.
[27] Poonam G,Sudha C,Nair,et al.Proteins with altered levels in plasma from Glioblastoma patients as revealed by iTRAQ-based quantitative proteomic analysis.PLoS One,2012,7(9):e46153-e46153.
[28] 王雅娟,刘进文,方向群,等.空间环境诱导褪色沙雷菌LCT-SM166的蛋白质组学分析.解放军医学院学报,2013,34(1):10-13.Wang Y J,Liu J W,Fang X Q,et al.Proteomics of space Serratia marcescens LCT-SM166 strain.Academic Journal of Pla Postgraduate Medical School,2013(1):10-13.

[1] 王光路, 王梦园, 周忆菲, 马科, 张帆, 杨雪鹏. 吡咯喹啉醌生物合成研究进展 *[J]. 中国生物工程杂志, 2021, 41(1): 103-113.
[2] 饶海密,梁冬梅,李伟国,乔建军,财音青格乐. 真菌芳香聚酮化合物的合成生物学研究进展*[J]. 中国生物工程杂志, 2020, 40(9): 52-61.
[3] 赵俊杰,张龙,王靓,陈旭升,毛忠贵. 具有双重抗生素抗性的ε-聚赖氨酸高产菌株选育及生理特性 *[J]. 中国生物工程杂志, 2018, 38(8): 59-68.
[4] 匙占库,文孟良,赵江源,李铭刚,韩秀林. 桉叶素生物合成研究进展[J]. 中国生物工程杂志, 2018, 38(11): 92-102.
[5] 沈伟涛, 王明钰, 王允坤, 王新华, 徐海. 抗生素含量测定方法的分析概述[J]. 中国生物工程杂志, 2016, 36(6): 119-126.
[6] 吕珊珊, 侯运华, 闫孟节, 钟耀华. 工业真菌高效产酶突变技术与高产机制[J]. 中国生物工程杂志, 2016, 36(3): 111-119.
[7] 王永成, 陈涛, 石婷, 王智文, 赵学明. 嘌呤核苷及其衍生物的代谢工程[J]. 中国生物工程杂志, 2015, 35(5): 87-95.
[8] 梁丽珠, 孙佳楠, 李恺, 刘明伟, 丁琛, 秦钧. 蛋白质组分析油酸对HepG2细胞转录因子DNA结合活性的影响[J]. 中国生物工程杂志, 2015, 35(5): 22-31.
[9] 李晓梅, 林春燕, 逄爱萍, 李晓波, 赵广荣. 合成生物学在链霉菌次级代谢产物研发中的应用[J]. 中国生物工程杂志, 2015, 35(4): 92-97.
[10] 王翠翠, 许蕙金兰, 傅达奇. 茄属生物碱的研究进展[J]. 中国生物工程杂志, 2015, 35(2): 99-104.
[11] 王洪秀, 张倩, 王玲杰, 唐科志. 链格孢菌毒素合成相关基因研究进展[J]. 中国生物工程杂志, 2015, 35(11): 92-98.
[12] 田宝玉, 马荣琴. 环境微生物的抗生素抗性和抗性组[J]. 中国生物工程杂志, 2015, 35(10): 108-114.
[13] 蒋延超, 蒋世云, 傅凤鸣, 黄凯, 康星欣, 徐丹. 透明质酸生物合成途径及基因工程研究进展[J]. 中国生物工程杂志, 2015, 35(1): 104-110.
[14] 张正玉, 吴绵斌. 抗生素分离纯化技术研究进展[J]. 中国生物工程杂志, 2012, 32(6): 98-103.
[15] 高燕会, 黄春红, 朱玉球, 童再康. 植物花青素苷生物合成及调控的研究进展[J]. 中国生物工程杂志, 2012, 32(08): 94-99.