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毛霉属对奶酪环境适应性反应的蛋白质组学分析。

Proteomic analysis of the adaptative response of Mucor spp. to cheese environment.

作者信息

Morin-Sardin Stéphanie, Jany Jean-Luc, Artigaud Sébastien, Pichereau Vianney, Bernay Benoît, Coton Emmanuel, Madec Stéphanie

机构信息

Université de Brest, EA 3882 Laboratoire Universitaire de Biodiversité et d'Ecologie Microbienne, IBSAM, ESIAB, Technopôle Brest-Iroise, 29280 Plouzané, France.

Université de Brest, UMR 6539, Laboratoire des Sciences de l'Environnement Marin, LEMAR CNRS/UBO/IRD/Ifremer, Institut Universitaire Européen de la Mer, Université de Bretagne Occidentale, 29280 Plouzané, France.

出版信息

J Proteomics. 2017 Feb 10;154:30-39. doi: 10.1016/j.jprot.2016.12.001. Epub 2016 Dec 6.

DOI:10.1016/j.jprot.2016.12.001
PMID:27940316
Abstract

UNLABELLED

In the cheese industry context, Mucor species exhibit an ambivalent behavior as some species are essential "technological" organisms of some cheeses while others can be spoiling agents. Previously, we observed that cheese "technological" species exhibited higher optimal growth rates on cheese related matrices than on synthetic media. This growth pattern combined with morphological differences raise the question of their adaptation to cheese. In this study, using a comparative proteomic approach, we described the metabolic pathways of three Mucor strains considered as "technological" or "contaminant" in the cheese environment (M. lanceolatus UBOCC-A-109153, M. racemosus UBOCC-A-109155, M. circinelloides CBS 277-49) as well as a non-cheese related strain (M. endophyticus CBS 385-95). Overall, 15.8 to 19.0% of the proteomes showed a fold change ≥1.6 in Potato Dextrose Agar (PDA) versus Cheese Agar (CA), a cheese mimicking-medium. The 289 differentially expressed proteins identified by LC MS-MS analysis were mostly assigned to energy and amino-acid metabolisms in PDA whereas a higher diversity of biological processes was observed for cheese related strains in CA. Surprisingly, the vast majority (72.9%) of the over-accumulated proteins were different according to the considered medium and strain. These results strongly suggest that the observed better adaptative response of "technological" strains to cheese environment is mediated by species-specific proteins.

BIOLOGICAL SIGNIFICANCE

The Mucor genus consists of a multitude of poorly known species. In the food context, few species are known for their positive role in the production of various food products, including cheese, while others are spoiling agents. The present study focused on the analysis of morphological and proteome differences of various Mucor spp. representative strains known as either positively (hereafter referred as "technological") or negatively (hereafter referred as "contaminant") associated with cheese or non-related to cheese (endophyte) on two different media, a synthetic medium and a cheese-mimicking medium. The main goal was to assess if adaptative traits of "technological" strains to the cheese environment could be identified. This work was based on observations we did in a recently published physiological study (Morin-Sardin et al., 2016). One of the important innovative aspects lies in the use for the first time of an extensive 2-DE approach to compare proteome variations for 4 strains on two different media. Results obtained offered an insight in the metabolic mechanisms associated with growth on a given medium and showed that adaptation to cheese environment is probably supported by species-specific proteins. The obtained data represent an essential step point for more targeted studies at the genomic and transcriptomic levels.

摘要

未标记

在奶酪行业背景下,毛霉属物种表现出矛盾的行为,因为一些物种是某些奶酪必不可少的“工艺”微生物,而另一些则可能是腐败菌。此前,我们观察到奶酪“工艺”物种在与奶酪相关的基质上比在合成培养基上表现出更高的最佳生长速率。这种生长模式与形态差异相结合,引发了它们对奶酪适应性的问题。在本研究中,我们采用比较蛋白质组学方法,描述了在奶酪环境中被视为“工艺”或“污染物”的三种毛霉菌株(披针毛霉UBOCC - A - 109153、总状毛霉UBOCC - A - 109155、卷枝毛霉CBS 277 - 49)以及一种与奶酪无关的菌株(内生毛霉CBS 385 - 95)的代谢途径。总体而言,在马铃薯葡萄糖琼脂(PDA)与模拟奶酪的奶酪琼脂(CA)培养基中,15.8%至19.0%的蛋白质组显示出≥1.6的倍数变化。通过液相色谱 - 质谱联用(LC MS - MS)分析鉴定出的289种差异表达蛋白在PDA中大多与能量和氨基酸代谢相关,而在CA中与奶酪相关的菌株观察到更高的生物过程多样性。令人惊讶的是,根据所考虑的培养基和菌株,绝大多数(72.9%)过度积累的蛋白质是不同的。这些结果强烈表明,观察到的“工艺”菌株对奶酪环境更好的适应性反应是由物种特异性蛋白质介导的。

生物学意义

毛霉属由众多鲜为人知的物种组成。在食品领域,很少有物种因其在包括奶酪在内的各种食品生产中的积极作用而闻名,而其他一些则是腐败菌。本研究重点分析了各种毛霉属代表性菌株在两种不同培养基(一种合成培养基和一种模拟奶酪的培养基)上的形态和蛋白质组差异,这些菌株与奶酪呈正相关(以下称为“工艺”)或负相关(以下称为“污染物”)或与奶酪无关(内生菌)。主要目标是评估是否可以识别“工艺”菌株对奶酪环境的适应特征。这项工作基于我们在最近发表的一项生理学研究(Morin - Sardin等人,2016年)中的观察结果。重要的创新点之一在于首次使用广泛的二维电泳(2 - DE)方法来比较4种菌株在两种不同培养基上的蛋白质组变化。获得的结果为与在给定培养基上生长相关的代谢机制提供了见解,并表明对奶酪环境的适应可能由物种特异性蛋白质支持。获得的数据代表了在基因组和转录组水平进行更有针对性研究的重要起点。

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