Shih Chao-Jen, Chen Sheng-Chung, Weng Chieh-Yin, Lai Mei-Chin, Yang Yu-Liang
Agricultural Biotechnology Research Center, Academia Sinica, Taipei, Taiwan.
Department of Life Sciences, National Chung Hsing University, Taichung, Taiwan;
Sci Rep. 2015 Nov 6;5:16326. doi: 10.1038/srep16326.
The aim of this study was to classify certain environmental haloarchaea and methanoarchaea using matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), and to expand the archaeal mass spectral database. A total of 69 archaea were collected including type strains and samples isolated locally from different environments. For extraction of the haloarchaeal total cell peptides/proteins, a simple method of acetonitrile extraction was developed. Cluster analysis conducted with the MALDI-TOF MS data overcame the high divergence in intragenomic 16S rRNA sequences in haloarchaea and clearly distinguished Methanohalophilus mahii from M. portucalensis. Putative biomarkers that can distinguish several particular archaeal genera were also assigned. In conclusion, this study expands the mass spectral database of peptide/protein fingerprints from bacteria and fungi to the archaea domain and provides a rapid identification platform for environmental archaeal samples.
本研究的目的是利用基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)对某些环境嗜盐古菌和甲烷古菌进行分类,并扩展古菌质谱数据库。共收集了69株古菌,包括模式菌株和从不同环境中本地分离的样本。为了提取嗜盐古菌的总细胞肽/蛋白质,开发了一种简单的乙腈提取方法。利用MALDI-TOF MS数据进行的聚类分析克服了嗜盐古菌基因组内16S rRNA序列的高度差异,并清楚地将马氏嗜甲烷菌与葡萄牙嗜甲烷菌区分开来。还确定了能够区分几个特定古菌属的推定生物标志物。总之,本研究将细菌和真菌的肽/蛋白质指纹质谱数据库扩展到了古菌领域,并为环境古菌样本提供了一个快速鉴定平台。