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高静水压对基质辅助激光解吸电离飞行时间质谱鉴定的影响——初步研究

Influence of High Hydrostatic Pressure on the Identification of by MALDI-TOF MS- Preliminary Study.

作者信息

Bucka-Kolendo Joanna, Sokołowska Barbara, Winiarczyk Stanisław

机构信息

Department of Microbiology, Institute of Agriculture and Food Biotechnology, 02-532 Warsaw, Poland.

Institute of High Pressure Physics, Polish Academy of Sciences, 01-142 Warsaw, Poland.

出版信息

Microorganisms. 2020 May 28;8(6):813. doi: 10.3390/microorganisms8060813.

Abstract

In the present study, we assessed the ability of MALDI-TOF MS (matrix-assisted laser desorption ionization time-of-flight mass spectrometry) to identify microbial strains subjected to high hydrostatic pressure (HHP) as a stress factor. Protein changes induced by HHP can affect the identification of microorganisms when the identification technique is based on the protein profile. We evaluated two methods, namely MALDI-TOF MS and 16S rDNA sequencing, as a valuable tool to identify species isolated from spoiled food, juices and beers. The data obtained from the protein mass fingerprint analysis of some of the lactobacilli strains showed differences in unpressured and pressured mass spectrum profiles (MSPs), which influenced the results of the identification. Four out of 13 strains (30%) showed different MSP results for unpressured and pressured samples and these results did not overlap with the 16S rDNA identification results. The 16S rDNA sequencing method revealed that five unpressured strains (38%) and four pressured strains (40%) were identified correctly by MALDI-TOF MS. Both methods showed compatible results in 38% of unpressured strains and in 30% of pressured strains. Stress factors, cultivation methods or the natural environment from which the bacteria were derived can affect their protein profile and thus change the mass spectrum. It is necessary to expand the database with a wide range of mass spectra dedicated to a high-throughput study of the microorganisms derived from different environments.

摘要

在本研究中,我们评估了基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)识别经受高静水压(HHP)这一应激因素的微生物菌株的能力。当识别技术基于蛋白质谱时,HHP诱导的蛋白质变化会影响微生物的识别。我们评估了两种方法,即MALDI-TOF MS和16S rDNA测序,作为识别从变质食品、果汁和啤酒中分离出的菌种的有价值工具。从一些乳酸杆菌菌株的蛋白质质量指纹分析获得的数据显示,未加压和加压质谱图(MSP)存在差异,这影响了识别结果。13株菌株中有4株(30%)未加压和加压样品的MSP结果不同,且这些结果与16S rDNA识别结果不重叠。16S rDNA测序方法显示,MALDI-TOF MS正确识别了5株未加压菌株(38%)和4株加压菌株(40%)。两种方法在38%的未加压菌株和30%的加压菌株中显示出一致的结果。应激因素、培养方法或细菌来源的自然环境会影响其蛋白质谱,从而改变质谱图。有必要用广泛的质谱图扩展数据库,以用于对来自不同环境的微生物进行高通量研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ef4e/7356497/baf7784182c3/microorganisms-08-00813-g001.jpg

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