Differential proteomics research of Bacillus amyloliquefaciens and its genome-shuffled saltant for improving fengycin production

Abstract In the previous study, we used genome shuffling to improve fengycin production of the original strain Bacillus amyloliquefaciens ES-2-4. After two rounds of genome shuffling, a high-yield recombinant FMB72 strain that exhibited 8.30-fold increase in fengycin production was obtained. In this study, comparative proteomic analysis of the parental ES-2-4 and genome-shuffled FMB72 strains was conducted to examine the differentially expressed proteins. In the shuffled strain FMB72, 50 differently expressed spots (p < 0.05) were selected to be excised and analyzed using Matrix-Assisted Laser Desorption/Ionization Time of Flight/Time of Flight Mass Spectrometry, and finally 44 protein spots were confidently identified according to NCBI database. According to clusters of orthologous groups (COG) functional category analysis and related references, the differentially expressed proteins could be classified into several functional categories, including proteins involved in metabolism, energy generation and conversion, DNA replication, transcription, translation, ribosomal structure and biogenesis, cell motility and secretion, signal transduction mechanisms, general function prediction. Of the 44 identified proteins, signaling proteins ComA and Spo0A may positively regulate fengycin synthesis at transcriptional level. Taken together, the present study will be informative for exploring the exact roles of ComA and Spo0A in fengycin synthesis and explaining the molecular mechanism of fengycin synthesis.

Kaempferol inhibits invasion and migration of non-small cell lung cancer A549 cells by down-regulating ERRα expression / 中国肿瘤生物治疗杂志

@# Objective: To study the effects of kaempferol on invasion and migration of non-small cell lung cancer (NSCLC)A549 cells and the related mechanisms. Methods: CCK-8 was used to detect the effect of different concentrations of kaempferol on the proliferation of A549 cells. Transwell assay and wound healing assay were used to detect the ability of cell invasion and migration. The expressions of EMT-related proteins (N-cadherin, Snail-2 and E-cadherin) were detected by Western blotting. The effect of kaempferol on the mRNA and protein expressions of estrogen related receptor alpha (ERRα) was determined by qRT-PCR. ERRα over-expression vector (pLV-ERRα) was transfected intoA549 cells, and the cell invasion, migration and the expression of EMT related proteins were detected by Transwell assay, wound healing assay and Western blotting, respectively. Results: Kaempferol dose-dependently inhibited the proliferation of A549 cells, and kaempferol at concentrations of 5, 10 and 20 μmol/L was used in the following experiments. After the treatment with different concentrations of kaempferol, the number of invasive cells and wound closure rate, the expression of N-cadherin and Snail-2, and the mRNAand protein levels of ERRα decreased significantly, while the expression of E-cadherin increased significantly, (all P<0.01). After transfection with pLV-ERRα, the number of invasive cells, scratch closure rate and the expressions of N-cadherin and Snail-2 of A549 cells over-expressing ERRα significantly increased, while the expression of E-cadherin decreased significantly (all P<0.05 or P<0.01); and all these indices were attenuated after the treatment with kaempferol (all P<0.05 or P<0.01). Conclusion: Kaempferol inhibits the invasion and migration as well as EMT of non-small cell lung cancer A549 cells by down-regulating ERRα, which may provide experimental basis for the clinical treatment of lung cancer.

Differential proteomics research of Bacillus amyloliquefaciens and its genome-shuffled saltant for improving fengycin production

Abstract In the previous study, we used genome shuffling to improve fengycin production of the original strain Bacillus amyloliquefaciens ES-24. After two rounds of genome shuffling, a high-yield recombinant FMB72 strain that exhibited 8.30-fold increase in fengycin production was obtained. In this study, comparative proteomic analysis of the parental ES-24 and genome-shuffled FMB72 strains was conducted to examine the differentially expressed proteins. In the shuffled strain FMB72, 50 differently expressed spots (p 0.05) were selected to be excised and analyzed using Matrix-Assisted Laser Desorption/Ionization Time of Flight/Time of Flight Mass Spectrometry, and finally 44 protein spots were confidently identified according to NCBI database. According to clusters of orthologous groups (COG) functional category analysis and related references, the differentially expressed proteins could be classified into several functional categories, including proteins involved in metabolism, energy generation and conversion, DNA replication, transcription, translation, ribosomal structure and biogenesis, cell motility and secretion, signal transduction mechanisms, general function prediction. Of the 44 identified proteins, signaling proteins ComA and Spo0A may positively regulate fengycin synthesis at transcriptional level. Taken together, the present study will be informative for exploring the exact roles of ComA and Spo0A in fengycin synthesis and explaining the molecular mechanism of fengycin synthesis.