Screening and Identification of Extracellular Protein Genes Relevant to Sulfur Activation of Extremely Thermoacidophilic Archaea Acidianus manzaensis

Abstract

Abstract: This study focused on extremely thermoacidophilic archaea Acidianus manzaensis，13 extracellular protein genes relevant to sulfur activation were screened and identified based on comparative proteomics，and their transcription levels were verified. First of all，extracellular proteins of A. manzaensis growing on elemental sulfur（S0）and ferrous（Fe2+）were extracted by slowly shaking water bath at 80℃ for 30 min，respectively. The proteins were separated by two-dimensional electrophoresis（2-DE）and differentially expressed spots under S0 substrate were selected. The selected spots were identified by tandem time of flight mass spectrometry（MALDI-TOF/TOF），and then analyzed using bioinformatics and functional prediction methods. Finally, the genes of the screened extracellular proteins associated with S0 activation were verified by real-time quantitative PCR（RT-qPCR）at transcription levels，and 13 relevant genes were acquired. More than half of the screened proteins contained plenty of cysteine residues（Cys），meaning thiol group（-SH）rich extracellular proteins participated in the process of S0 activation. Especially，both glutaredoxin and FAD-linked oxidase contained -CXXC- domain，and their corresponding genes were highly expressed，indicating that the proteins with -CXXC- domain play an important role during the S0 activation process of extremely thermoacidophilic archaea A. manzaensis.

Key words: Acidianus manzaensis, comparative proteomics, extracellular protein, sulfur activation

CLC Number:

10.13560/j.cnki.biotech.bull.1985.2016.12.023

MA Ya-long,LIU Hong-chang,XIA Jin-lan，YANG Yun ,NIE Zhen-yuan,. Screening and Identification of Extracellular Protein Genes Relevant to Sulfur Activation of Extremely Thermoacidophilic Archaea Acidianus manzaensis[J]. Biotechnology Bulletin, 2016, 32(12): 143-151.

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