以核糖体蛋白质鉴别铜绿微囊藻的应用分析

doi:10.13560/j.cnki.biotech.bull.1985.2017-0268

生物技术通报 ›› 2017, Vol. 33 ›› Issue (10): 117-124.doi: 10.13560/j.cnki.biotech.bull.1985.2017-0268

以核糖体蛋白质鉴别铜绿微囊藻的应用分析

江文静^{1, 2}, 张军毅², 杜阳^{1, 2}, 孙丽伟^{1, 2}

1. 东南大学能源与环境学院东南大学太湖水环境工程研究中心，无锡 214000
2. 无锡市环境监测中心站，无锡 214000

收稿日期:2017-04-05 出版日期:2017-10-29 发布日期:2017-10-29
作者简介:江文静，女，硕士研究生，研究方向：鉴别蓝藻新方法的研究与应用；E-mail：1452765330@qq.com
基金资助:
国家重点研发计划（2016YFC0400801）

Application Analysis of Microcystis aeruginosa Identification Based on Ribosomal Proteins

JIANG Wen-jing^1，2, ZHANG Jun-yi², DU Yang^1，2, SUN Li-wei^1，2

1. School of Energy and Environment，Southeast University，Taihu Lake Water Environment Engineering Research Center，Southeast University，Wuxi 214000
2. Wuxi Environmental Monitoring Centre，Wuxi 214000

Received:2017-04-05 Published:2017-10-29 Online:2017-10-29

摘要/Abstract

摘要： 旨为研究以核糖体蛋白质为生物标识物的基质辅助激光解吸/电离平衡飞行时间质谱（MALDI-TOF MS）法鉴别混合及实际蓝藻样品的可行性。通过机械破碎和高速离心的前处理步骤获取待测蓝藻样品的核糖体蛋白质组分，使蛋白质原位结晶后进行MALDI-TOF MS测定，并选择特定的13个核糖体蛋白质为生物标识物进行蛋白质组学分析。结果表明在筛选的最优实验条件下，模式藻株铜绿微囊藻（Microcystis aeruginosa）能在混合样品中被精确鉴别出，标识物特征峰的检出率均大于75%，通过对不同藻细胞密度的样品测定，混合蓝藻样品中M. aeruginosa的最低生物量检出限为2.88×10^{6cell，所占比例为37%，核糖体蛋白质特征峰值的误差在合理范围内；M. aeruginosa在太湖实际蓝藻样品中也被成功鉴别出，特征峰的检出率为76.9%。综合分析表明，基于核糖体蛋白质为生物标识物的MALDI-TOF MS法能成功应用于混合和实际蓝藻样品的鉴别且可能成为未来实际水体中蓝藻日常监测的手段。}

关键词: 核糖体蛋白质, 铜绿微囊藻, MALDI-TOF MS, 蓝藻水华

Abstract: This research is to verify the feasibility of matrix-assisted laser desorption-ionization-time-of -flight mass spectrometry（MALDI-TOF MS）in the identification of mixed and actual cyanobacterial samples by employing ribosomal proteins as biomarkers. Mechanical disruption and high speed centrifugation were used as pre-treatments to collect the ribosomal protein fraction of cyanobacterial samples，and then MALDI-TOF MS was applied to identify Microcystis aeruginosa in samples after proteins in situ crystallization with proteomic analysis based on 13 ribosomal proteins as biomarkers. The results showed that under the optimal conditions，model strain M. aeruginosa was precisely identified in the mixed samples and the detection rates of assigned peaks were all greater than 75% with reasonable errors range except the fourth sample. The lowest biomass detection limit and ratio of M. aeruginosa was 2.88×10^{6 cells and 37%，respectively. Furthermore，M. aeruginosa was identified and detected in the cyanobacterial samples from Taihu Lake and the detection rate of assigned peaks was 76.9%. In conclusion，MALDI-TOF MS employing ribosomal proteins as biomarkers could be successfully applied to the identification of mixed and actual cyanobacterial samples and will be expected as routine tool in environmental cyanobacteria monitoring.}

Key words: ribosomal protein, Microcystis aeruginosa, MALDI-TOF MS, cyanobacterial bloom

江文静, 张军毅, 杜阳, 孙丽伟. 以核糖体蛋白质鉴别铜绿微囊藻的应用分析[J]. 生物技术通报, 2017, 33(10): 117-124.

JIANG Wen-jing, ZHANG Jun-yi, DU Yang, SUN Li-wei. Application Analysis of Microcystis aeruginosa Identification Based on Ribosomal Proteins[J]. Biotechnology Bulletin, 2017, 33(10): 117-124.

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以核糖体蛋白质鉴别铜绿微囊藻的应用分析

Application Analysis of Microcystis aeruginosa Identification Based on Ribosomal Proteins

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