Plumridge Andrew, Hesse Stephan J A, Watson Adrian J, Lowe Kenneth C, Stratford Malcolm, Archer David B
School of Biology, University of Nottingham, Nottingham, NG7 2RD, United Kingdom.
Appl Environ Microbiol. 2004 Jun;70(6):3506-11. doi: 10.1128/AEM.70.6.3506-3511.2004.
The growth of the filamentous fungus Aspergillus niger, a common food spoilage organism, is inhibited by the weak acid preservative sorbic acid (trans-trans-2,4-hexadienoic acid). Conidia inoculated at 10(5)/ml of medium showed a sorbic acid MIC of 4.5 mM at pH 4.0, whereas the MIC for the amount of mycelia at 24 h developed from the same spore inoculum was threefold lower. The MIC for conidia and, to a lesser extent, mycelia was shown to be dependent on the inoculum size. A. niger is capable of degrading sorbic acid, and this ability has consequences for food preservation strategies. The mechanism of action of sorbic acid was investigated using (31)P nuclear magnetic resonance (NMR) spectroscopy. We show that a rapid decline in cytosolic pH (pH(cyt)) by more than 1 pH unit and a depression of vacuolar pH (pH(vac)) in A. niger occurs in the presence of sorbic acid. The pH gradient over the vacuole completely collapsed as a result of the decline in pH(cyt). NMR spectra also revealed that sorbic acid (3.0 mM at pH 4.0) caused intracellular ATP pools and levels of sugar-phosphomonoesters and -phosphodiesters of A. niger mycelia to decrease dramatically, and they did not recover. The disruption of pH homeostasis by sorbic acid at concentrations below the MIC could account for the delay in spore germination and retardation of the onset of subsequent mycelial growth.
丝状真菌黑曲霉是一种常见的食品腐败微生物,其生长受到弱酸防腐剂山梨酸(反式-反式-2,4-己二烯酸)的抑制。在培养基中接种浓度为10(5)/ml的分生孢子,在pH 4.0时山梨酸的最低抑菌浓度(MIC)为4.5 mM,而由相同孢子接种物在24小时内发育的菌丝体数量的MIC则低三倍。分生孢子以及在较小程度上菌丝体的MIC显示取决于接种量。黑曲霉能够降解山梨酸,这种能力对食品保鲜策略有影响。使用(31)P核磁共振(NMR)光谱研究了山梨酸的作用机制。我们发现,在山梨酸存在的情况下,黑曲霉的胞质pH(pH(cyt))迅速下降超过1个pH单位,液泡pH(pH(vac))降低。由于pH(cyt)的下降,液泡上的pH梯度完全消失。NMR光谱还显示,山梨酸(在pH 4.0时为3.0 mM)导致黑曲霉菌丝体的细胞内ATP池以及糖磷酸单酯和磷酸二酯水平急剧下降,且无法恢复。低于MIC浓度的山梨酸对pH稳态的破坏可能是孢子萌发延迟和随后菌丝体生长起始延迟的原因。