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解析福山假丝酵母RCL-3本土酵母中与铜抗性相关的代谢途径。

Disentangling metabolic pathways involved in copper resistance in Candida fukuyamaensis RCL-3 indigenous yeast.

作者信息

Irazusta Verónica, Michel Lucas, de Figueroa Lucía I C

机构信息

PROIMI-CONICET, Tucumán, Argentina.

INIQUI-CONICET-UNSa, Salta, Argentina.

出版信息

J Basic Microbiol. 2016 Jul;56(7):698-710. doi: 10.1002/jobm.201500509. Epub 2015 Nov 16.

Abstract

Candida fukuyamaensis RCL-3 yeast strain isolated from a copper filter plant is able to lower copper concentration in culture medium. In the present study, effect of copper in proteins expression and mechanisms involved in copper resistance were explored using comparative proteomics. Mono-dimensional gel electrophoresis revealed differential band expressions between cells grown with or without copper. 2-DE analysis of C. fukuyamaensis RCL-3 revealed that copper exposure produced at least an over-expression of 40 proteins. Sixteen proteins were identified and grouped in four categories according to their functions: glycolysis and ATP production, synthesis of proteins, oxidative stress response, and processing and transport of proteins. Integral membrane proteins and membrane-associated proteins were analyzed, showing nine protein bands over-expressed in Cu-supplemented medium. Four proteins were identified, namely nucleoporin pom152, elongation factor 2, copper chaperone Sod1 Ccs1, and eiosome component Lsp1. The proteomic analysis performed allowed the identification of different metabolic pathways and certain proteins involved in metal input and storage related to cell ability to bioremediate copper. These proteins and mechanisms could be used for future applications of C. fukuyamaensis RCL-3 in biotechnological processes such as remediation of heavy metals.

摘要

从一家铜过滤厂分离出的福山假丝酵母RCL - 3酵母菌株能够降低培养基中的铜浓度。在本研究中,利用比较蛋白质组学探究了铜对蛋白质表达的影响以及铜抗性相关机制。一维凝胶电泳显示了在有铜和无铜条件下生长的细胞之间条带表达的差异。对福山假丝酵母RCL - 3进行的二维电泳分析表明,铜暴露至少使40种蛋白质过度表达。鉴定出16种蛋白质,并根据其功能分为四类:糖酵解和ATP生成、蛋白质合成、氧化应激反应以及蛋白质加工和运输。对整合膜蛋白和膜相关蛋白进行了分析,结果显示在添加铜的培养基中有9条蛋白条带过度表达。鉴定出了4种蛋白质,即核孔蛋白pom152、延伸因子2、铜伴侣Sod1 Ccs1和脂筏成分Lsp1。所进行的蛋白质组学分析使得能够鉴定出不同的代谢途径以及与细胞生物修复铜能力相关的参与金属输入和储存的某些蛋白质。这些蛋白质和机制可用于福山假丝酵母RCL - 3未来在生物技术过程(如重金属修复)中的应用。

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