In situ investigation of Geobacillus stearothermophilus spore germination and inactivation mechanisms under moderate high pressure

被引:30
|
作者
Georget, Erika [1 ]
Kapoor, Shobhna [2 ]
Winter, Roland [2 ]
Reineke, Kai [4 ]
Song, Youye [1 ]
Callanan, Michael [3 ]
Ananta, Edwin [3 ]
Heinz, Volker [1 ]
Mathys, Alexander [1 ]
机构
[1] German Inst Food Technol DIL, D-49610 Quakenbruck, Germany
[2] Tech Univ Dortmund, Fak Chem & Chem Biol, D-44227 Dortmund, Germany
[3] Nestle Res Ctr, CH-1000 Lausanne, Switzerland
[4] Leibniz Inst Agr Engn Potsdam Bornim eV, D-14469 Potsdam, Germany
关键词
Bacterial spore; High pressure; Germination; In situ; Fluorescence; FT-IR spectroscopy; BACILLUS-SUBTILIS; MEMBRANE-FLUIDITY; BACTERIAL-SPORES; INNER MEMBRANE; DORMANT SPORES; LAURDAN; TEMPERATURE; RESISTANCE; PROTEINS; NUTRIENT;
D O I
10.1016/j.fm.2014.01.007
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Bacterial spores are a major concern for food safety due to their high resistance to conventional preservation hurdles. Innovative hurdles can trigger bacterial spore germination or inactivate them. In this work, Geobacillus stearothermophilus spore high pressure (HP) germination and inactivation mechanisms were investigated by in situ infrared spectroscopy (FT-IR) and fluorometry. G. stearothermophilus spores' inner membrane (IM) was stained with Laurdan fluorescent dye. Time-dependent FT-IR and fluorescence spectra were recorded in situ under pressure at different temperatures. The Laurdan spectrum is affected by the lipid packing and level of hydration, and provided information on the IM state through the Laurdan generalized polarization. Changes in the -CH2 and -CH3 asymmetric stretching bands, characteristic of lipids, and in the amide I' band region, characteristic of proteins' secondary structure elements, enabled evaluation of the impact of HP on endospores lipid and protein structures. These studies were complemented by ex situ analyses (plate counts and microscopy). The methods applied showed high potential to identify germination mechanisms, particularly associated to the IM. Germination up to 3 log(10) was achieved at 200 MPa and 55 degrees C. A molecular-level understanding of these mechanisms is important for the development and validation of multi-hurdle approaches to achieve commercial sterility. (C) 2014 Elsevier Ltd. All rights reserved.
引用
收藏
页码:8 / 18
页数:11
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