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比较蛋白质组学揭示肉类腐败菌的厌氧生活方式。

Comparative Proteomics Reveals the Anaerobic Lifestyle of Meat-Spoiling Species.

作者信息

Kolbeck Sandra, Abele Miriam, Hilgarth Maik, Vogel Rudi F

机构信息

Lehrstuhl für Technische Mikrobiologie, Technische Universität München, Freising, Germany.

Bayerisches Zentrum für Biomolekulare Massenspektrometrie (BayBioMS), Freising, Germany.

出版信息

Front Microbiol. 2021 Apr 6;12:664061. doi: 10.3389/fmicb.2021.664061. eCollection 2021.

Abstract

The ability of certain () species to grow or persist in anoxic habitats by either denitrification, acetate fermentation, or arginine fermentation has been described in several studies as a special property. Previously, we had isolated strains belonging to the species , , and from anoxic modified atmosphere packaged (MAP) minced beef and further proved their anaerobic growth on agar plates. This follow-up study investigated the anaerobic growth of two strains per respective species on inoculated chicken breast filet under 100% N modified atmosphere. We were able to prove anaerobic growth of all six strains on chicken breast filet with cell division rates of 0.2-0.8/day. Furthermore, we characterized the anaerobic metabolic lifestyle of these strains by comparative proteomics, upon their cultivation in meat simulation media, which were constantly gassed with either air or 100% N atmospheres. From these proteomic predictions, and respective complementation by physiological experiments, we conclude that the strains , , exhibit a similar anaerobic lifestyle and employ arginine fermentation via the arginine deiminase (ADI) pathway to grow anaerobically also on MAP meats. Furthermore, glucose fermentation to ethanol via the ED-pathway is predicted to enable long term survival but no true growth, while respiratory growth with nitrate as alternative electron acceptor or glucose fermentation to acetate could be excluded due to absence of essential genes. The citric acid cycle is partially bypassed by the glyoxylate shunt, functioning as the gluconeogenetic route without production of NADH under carbon limiting conditions as e.g., in packaged meats. Triggered by an altered redox balance, we also detected upregulation of enzymes involved in protein folding as well as disulfide bonds isomerization under anoxic conditions as a counteracting mechanism to reduce protein misfolding. Hence, this study reveals the mechanisms enabling anaerobic grow and persistence of common meat-spoiling species, and further complements the hitherto limited knowledge of the anaerobic lifestyle of species in general.

摘要

某些()物种通过反硝化作用、乙酸发酵或精氨酸发酵在缺氧生境中生长或存活的能力,在多项研究中被描述为一种特殊特性。此前,我们已从缺氧气调包装(MAP)的碎牛肉中分离出属于、和物种的菌株,并进一步证明了它们在琼脂平板上的厌氧生长。这项后续研究调查了每个物种的两株菌株在100%氮气调气氛下接种的鸡胸肉排上的厌氧生长情况。我们能够证明所有六株菌株在鸡胸肉排上厌氧生长,细胞分裂速率为0.2 - 0.8/天。此外,我们通过比较蛋白质组学对这些菌株在肉模拟培养基中培养时的厌氧代谢生活方式进行了表征,培养基持续用空气或100%氮气气氛充气。从这些蛋白质组学预测以及生理实验的相应补充中,我们得出结论,菌株、、表现出相似的厌氧生活方式,并通过精氨酸脱亚氨酶(ADI)途径利用精氨酸发酵,从而也能在MAP肉类上厌氧生长。此外,预计通过ED途径将葡萄糖发酵为乙醇可实现长期存活,但不能真正生长,而由于缺乏必需基因,可排除以硝酸盐作为替代电子受体的呼吸生长或葡萄糖发酵为乙酸的情况。柠檬酸循环部分被乙醛酸循环旁路绕过,并在碳限制条件下(如包装肉类中)作为糖异生途径发挥作用,且不产生NADH。受氧化还原平衡改变的触发,我们还检测到缺氧条件下参与蛋白质折叠以及二硫键异构化的酶上调,这是一种减少蛋白质错误折叠的抵消机制。因此,本研究揭示了常见肉类腐败()物种实现厌氧生长和存活的机制,并进一步补充了迄今为止对()物种厌氧生活方式总体上有限的认识。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5335/8055858/b67ba63351ae/fmicb-12-664061-g001.jpg

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