| Arthrospira is one of the most cultivated commercial microalgae in the world,and its high yield,high quality,high efficiency,low cost cultivation technologies have always been in hot research.Study on the influencing factors and regulation mechanism of the production traits could have great significance to the breeding and achievement of high yield and low cost of productionin Arthrospira.5 good production traits strains including ZJU-0103,ZJU-0105,ZJU-0107,ZJU-0108,ZJU-0115 and 3 poor production traits strains including ZJU-0101,ZJU-0110,ZJU-0114 were selected in the current study.Commom significant differences could be observed at 102 kD between the above strains by SDS-PAGE analysis of water-soluble proteins.We considered that the proteins at 102 kD could have relation to the production traits of Arthrospira(PTP102).Furthermore,the kinds and preliminary characteristics of PTP102 were identified and analyzed,and the results were as follows:1.Mass spectrometry and bioinformatics analysis of PTP102 from ArthrospiraThe PTP102 of ZJU-0101 and ZJU-0103 were identified by LC-MS/MS,and the results showed:(1)27 different proteins were identified,and 22 of which were up-regulated in ZJU-0103,while the other 5 were up-regulated in ZJU-0101.(2)The molecular weight comparison between the theoretical values and estimated values by SDS-PAGE of PTP102 showed that only 9 proteins were below the ratio of 10%,including Phycobilisome core-membrane linker polypeptide,Helicase,Valine--tRNA ligae,Peptidase M1 membrane alanine aminopeptidase,DNA mismatch repair protein MutS,Protein translocase subunit SecA and 3 Putative uncharacterized protein.(3)Subcellular localization analysis of the 22 known proteins showed that cytoplasmic protein accounted for 77%,extracellular protein accounted for 18%,periplasmic protein accounted for 5%.(4)Furthermore,these proteins could be classified into 7 categories based on their functions,in which synthesis and transport-related proreins accounted for 27%,photosynthesis-related proreins accounted for 18%,environmental adaptation-related proreins accounted for 18%,nucleotide metabolism-related proreins accounted for 14%,energy metabolism related proreins accounted for 9%,material hydrolysis-related proreins accounted for 9%,and proteins involved in material absorbtion and exocytosis accounted for 4%.2.Correlation analysis between PTP102 and production traits based on Athrospira genome sequencesThe production traits of 7 Arthrospira strains whose genome sequences were published base on the established molecular markers of 16-23S rRNA ITS,cpcH,cpcI,cpcD,mreB and ftsZ.Then the relationship between PTP102 and production traits of Arthrospira were investigated based on the gene sequences,and the result were as follows:(1)Arthrospira platensis C1,Arthrospira platensis PCC8005,Arthrospira platensis TJSD091and Arthrospira maxima CS-328 were predicted to be poor traits stains,while Arthrospira platensis NIES-39,Arthrospira platensis YZ and Arthrospira platensis Paraca were predicted to be good traits stains.(2)Base on the phylogynetic trees built by the 27 proteins,we found that CpcH,CpcI,Phosphoglycerate kinase,Valine-tRNA ligase,Helicase,Protein translocase subunit SecA,Elongation factor Tu,Elongation factor Ts,DNA mismatch repair protein MutS,Signal recognition particle receptor FtsY and Peptidase M1 membrane alanine aminopeptidase could divide the Arthrospira strains into two clusters.(3)The amino acid sequences of the above proteins were compared by multiple sequences analysis,and the result indicated that there were common differential sits among these Arthrospira strains,which further revealed the correlation between PTP102 and Arthrospira strains.(4)Based on the analysis of the molecular weight of PTP102 and its phylogenetic tree,we found that valine-tRNA ligase,helicase,protein transcriptase subunit SecA,DNA mismatch repair protein MutS and M1 peptidase Proteins were closely related to the production traits of Arthrospira.3.Analysis of the preliminary characteristics of PTP102 from ArthrospiraThe stabilities of PTP102 from ZJU-0101 and ZJU-0103 at different temperatures were analyzed,as well as their isoelectric points.The results showed that PTP102 remained stable below the temperature of 30 0C.Meanwhile,two-dimensional electrophoresis analysis showed that the isoelectric points of PTP102 distributed in 4-5,which was consistent with the result by LC-MS/MS.All the results contributed to further research of separation,purification and identification of PTP 102. |
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