Luu Stephanie, Cruz-Mora Jose, Setlow Barbara, Feeherry Florence E, Doona Christopher J, Setlow Peter
Department of Molecular Biology and Biophysics, UConn Health, Farmington, Connecticut, USA.
U.S. Army-Natick Soldier RD&E Center, Warfighter Directorate, Natick, Massachusetts, USA.
Appl Environ Microbiol. 2015 Apr;81(8):2927-38. doi: 10.1128/AEM.00193-15. Epub 2015 Feb 13.
Nutrient germination of spores of Bacillus species occurs through germinant receptors (GRs) in spores' inner membrane (IM) in a process stimulated by sublethal heat activation. Bacillus subtilis spores maximum germination rates via different GRs required different 75 °C heat activation times: 15 min for l-valine germination via the GerA GR and 4 h for germination with the L-asparagine-glucose-fructose-K(+) mixture via the GerB and GerK GRs, with GerK requiring the most heat activation. In some cases, optimal heat activation decreased nutrient concentrations for half-maximal germination rates. Germination of spores via various GRs by high pressure (HP) of 150 MPa exhibited heat activation requirements similar to those of nutrient germination, and the loss of the GerD protein, required for optimal GR function, did not eliminate heat activation requirements for maximal germination rates. These results are consistent with heat activation acting primarily on GRs. However, (i) heat activation had no effects on GR or GerD protein conformation, as probed by biotinylation by an external reagent; (ii) spores prepared at low and high temperatures that affect spores' IM properties exhibited large differences in heat activation requirements for nutrient germination; and (iii) spore germination by 550 MPa of HP was also affected by heat activation, but the effects were relatively GR independent. The last results are consistent with heat activation affecting spores' IM and only indirectly affecting GRs. The 150- and 550-MPa HP germinations of Bacillus amyloliquefaciens spores, a potential surrogate for Clostridium botulinum spores in HP treatments of foods, were also stimulated by heat activation.
芽孢杆菌属孢子的营养萌发通过孢子内膜(IM)中的萌发受体(GRs)发生,该过程由亚致死热激活刺激。枯草芽孢杆菌孢子通过不同GRs的最大萌发率需要不同的75°C热激活时间:通过GerA GR进行L-缬氨酸萌发需要15分钟,通过GerB和GerK GRs用L-天冬酰胺-葡萄糖-果糖-K(+)混合物萌发需要4小时,其中GerK需要的热激活最多。在某些情况下,最佳热激活会降低达到半数最大萌发率所需的营养物浓度。通过150 MPa的高压(HP)使孢子通过各种GRs萌发,其热激活要求与营养萌发相似,并且最佳GR功能所需的GerD蛋白缺失并未消除达到最大萌发率的热激活要求。这些结果与热激活主要作用于GRs一致。然而,(i)如通过外部试剂进行生物素化所探测的,热激活对GR或GerD蛋白构象没有影响;(ii)在影响孢子IM特性的低温和高温下制备的孢子,在营养萌发的热激活要求上表现出很大差异;(iii)550 MPa的HP使孢子萌发也受热激活影响,但这些影响相对独立于GRs。最后的结果与热激活影响孢子的IM且仅间接影响GRs一致。解淀粉芽孢杆菌孢子(在食品的HP处理中可作为肉毒梭菌孢子的潜在替代物)的150 MPa和550 MPa HP萌发也受热激活刺激。