The Purification And Identification Of Copper Binding Protein From Penicillium Janthinellum GXCR

By using Penicillium janthinellum GXCR which has been screenedbefore as material, the experiment was carried out to obtain proteins aboutcopper resistance by separated and identified copper binding proteins fromPenicillium janthinellum GXCR with copper ion inducing or without copperion inducing respectively.The proteins were extracted from Penicillium janthinellum GXCRwithout copper ions inducing. A series of copper binding proteins werepurified by copper affinity chromatography. 17 copper binding proteins wererandom selected and analysed by matrix-assisted laser desorption ionizationtime-of-flight mass spectrometry. A summary of the identified proteins arepresented. The profile of peptide mass fingerprinting of 12 proteins weresubmitted in SWISS-PROT database and NCBI database, and the resultsrevealed that they showed higher homology with thses following proteins:NADP-specific glutamate dehydrogenase, E3 ubiquitin protein ligase TOM1,ubiquitin peptide,histidyl-tRNA synthetase,NADP linked glutamate dehydrogenase,aspartate aminotransferase,metaxin 1 homolog, SHE 4 proteinand 4 hypothetical proteins. Up to now the functions of metaxin 1 homolog,SHE 4 protein, and 4 hypothetical protein are still unknow. There are noreports about these proteins response to heavy metal resistance of microbes.The function of these proteins for the heavy metal resistance need to be studying more.The copper binding protein from Penieillium janthinellum GXCR undercopper ions inducing was isolated by Sephadex G-75 and Cellulose DEAE-52.The analysis of SDS-PAGE electrophoresis revealed that the MW of purifiedcopper binding protein was approximate 20kDa.This protein was identified using peptide mass fingerprinting andmatrix-assisted laser desorption ionization time-of-flight mass spectrometry.No matched protein was found in the SWISS-PROT database and NCBIdatabase, so it is worth further studying.