Comparative Proteomic Analysis Of Streptomyces Roseoflavus Men-myco-93-63 And Its Mutants

Streptomyces roseoflavus Men-myco-93-63 is an antagonistic strain isolated from potato scab declined soil, the strain and it's fermentation has strong inhibition to many phytopathogenic fungi such as cotton Verticillium wilt caused by Verticillium dahliae, cumber powdery mildew caused by Sphaerotheca fuliginea Poll, and so on, thus showed great application potential in bio-control field.Comparative proteomic analysis of Streptomyces roseoflavus Men-myco-93-63 and its mutants, including its blocked mutants MV11, MV18, MV21, MV22, MV35, MV40, DE01, DE02 and high yield mutant D12-3 were employed in this research. Total protein were isolated and analyzed by two-dimensional gel electrophoresis. Different expression protein spots were acquired and analyzed by Peptide Mass Fingerprinting (PMF). These may provide theoretical basis to understand the biosynthesis of antibiotic.(1) Two-dimensional electrophoresis system suitable to Streptomyces roseoflavus was developed. The results showed that the majority proteins of Streptomyces were acidic and distributed from pH4 to pH7.(2) Differentially expressed proteins between wild strain and blocked mutants were analyzed, and two protein spots were identified, one of them showed high similarity with hypothetical protein of Synechococcus sp.. Another one showed similarity with GroES of Streptomyces Coelicolor, it may be involved in energy metabolism during mutagenesis to ensure Nascent Peptide's synthesis, folding, and the configuration adjustment.(3) Two protein spots from MV35 were identified in the co-synthesis analysis. L351 showed remarkable some similarity with ectoine hydroxylase, it may be regulated by secretion of coverner MV35 positively. L353 showed similarity with transcriptional regulator. The corresponding coding gene may be related with synthesis of material secreted by MV35, which conduced positive regulation to the antibiotic synthesis .(4) Differentially expressed proteins between wild strain and high yield were analyzed, and four protein spots were identified. L12-1 showed some remarkable similarity with PhaP, which can control the synthesis of polyhydroxyalkanoates by regulating the expression of PhaC, it may be related with the metabolic pathway of antibiotic. L12-2 showed remarkable some similarity withβ-lactamase, which could makeβ-lactam antibiotic lost its resistance by degrading beta-lactam loop, it was presumed that this protein linked with synthetic gene of antibiotic. L12-3 showed similarity with methyl-accepting chemotaxis sensory transducer. It was receptor protein responsibility to induce changes of environment. In this research, it perhaps produced by mutating to wild strain. Relationship of this gene and metabolic pathway of antibiotic need to be further studied. L93-4 showed some similarity with ATP-binding protein that could bind with ATP and be involved in the transportation of intra- and extracellular materials, which assistance metabolism of strain thalli cell.