Peptide-Mediated Gene Transfer into Marine Purple Photosynthetic Bacteria

• Published in: International journal of molecular sciences 2020-11, Vol.21 (22), p.8625
• Main Authors: Higuchi-Takeuchi, Mieko, Miyamoto, Takaaki, Foong, Choon Pin, Goto, Mami, Morisaki, Kumiko, Numata, Keiji
• Format: Article
• Language: English
• Subjects: Actin; Amino acids; Aquatic Organisms - genetics; Aquatic Organisms - metabolism; Bacteria; Biotechnology; cell penetrating peptide; Cell-Penetrating Peptides - chemistry; Chloride; Conjugation; D-Amino acids; Deoxyribonucleic acid; DNA; Efficiency; Escherichia coli - genetics; Escherichia coli - metabolism; Gene transfer; Gene Transfer Techniques; Genetic transformation; Growth stage; Homology; Microorganisms; Nanomaterials; Peptides; Photosynthesis; plasmid DNA delivery; Plasmids; Plasmids - chemistry; Plasmids - genetics; Proteins; Rhodobacteraceae - genetics; Rhodobacteraceae - metabolism; Rhodovulum sulfidophilum; Sarcosine; transformation; Transformations
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms21228625

Use of photosynthetic organisms is one of the sustainable ways to produce high-value products. Marine purple photosynthetic bacteria are one of the research focuses as microbial production hosts. Genetic transformation is indispensable as a biotechnology technique. However, only conjugation has been determined to be an applicable method for the transformation of marine purple photosynthetic bacteria so far. In this study, for the first time, a dual peptide-based transformation method combining cell penetrating peptide (CPP), cationic peptide and Tat-derived peptide (dTat-Sar-EED) (containing D-amino acids of Tat and endosomal escape domain (EED) connected by sarcosine linkers) successfully delivered plasmid DNA into a marine purple photosynthetic bacterium. The plasmid delivery efficiency was greatly improved by dTat-Sar-EED. The concentrations of dTat-Sar-EED, cell growth stage and recovery duration affected the efficiency of plasmid DNA delivery. The delivery was inhibited at 4 °C and by A22, which is an inhibitor of the actin homolog MreB. This suggests that the plasmid DNA delivery occurred via MreB-mediated energy dependent process. Additionally, this peptide-mediated delivery method was also applicable for cells. Thus, a wide range of bacteria could be genetically transformed by using this novel peptide-based transformation method.