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Production of recombinant colicin M in Nicotiana tabacum plants and its antimicrobial activity
Ewelina Łojewska,Tomasz Sakowicz,Aleksandra Kowalczyk,Magdalena Konieczka,Janina Grzegorczyk,Przemysław Sitarek,Ewa Skała,Piotr Czarny,Tomasz Śliwiński,Tomasz Kowalczyk 한국식물생명공학회 2020 Plant biotechnology reports Vol.14 No.1
Antibiotic-resistant microorganisms causing a life-threatening infection pose a serious challenge for modern science. The rapidly growing number of incidents for which the use of standard antibiotics is ineffective forces us to develop new alternative methods of killing microorganisms. Antimicrobial proteins and peptides (AMPs) can be promising candidates to solve this problem. Colicin-M is one of the representatives of this group and is naturally produced by Escherichia coli acting on other closely related bacterial strains by disrupting their outer cell membrane. This bacteriocin has huge potential as a potent antimicrobial agent, especially, since it was recognized by the FDA as safe. In this work, we present the expression of colicin M in model transgenic Nicotiana tabacum L. plants. We demonstrate that purified colicin retains its antibacterial activity against the control Escherichia coli strains and clinical isolates of Escherichia coli and Klebsiella pneumoniae. Our results also show that plant-derived ColM is not toxic for L929 and HeLa cell line, which allows us to suppose that plant-based expression could be an alternative production method of such important proteins.
( Radoslaw Sikora ),( Elzbieta Grzesiuk ),( Sikora Anna ),( Justyna Wojtowicz Sienko ),( Piotr Piela ),( Urszula Zielenkiewicz ),( Karolina Tomczyk Zak ),( Aleksandra Chojnacka ),( Pawe Kowalczyk ),( 한국미생물 · 생명공학회 2011 Journal of microbiology and biotechnology Vol.21 No.3
Ferric ion-respiring microorganisms (FRMs) are a group of prokaryotes that use Fe(III) as well as other metals as terminal electron acceptors in the process of anaerobic respiration. Special attention is paid to a biotechnological significance of FRMs because of their potential role in electricity production in microbial fuel cells (MFCs) where the terminal acceptor of the electrons during anaerobic respiration is not a ferric ion but the anode. One of the best known FRMs is the Shewanellaceae family. Most of the Shewanella species have been isolated from marine environments. In this report, sugar beet molasses and ferric oxide were successfully used in the selection of a bacterial consortium capable of dissimilatory Fe(III) reduction in a long-term continuous culture. The inoculum was a sample of eutrophic lake bottom sediment. Among the bacteria present in this culture were representatives of the Enterobacteriaceae, and the genera Pseudomonas, Arcobacter, and Shewanella. Two non-marine Fe(III)-reducing Shewanella-related clones named POL1 and POL2 were isolated. The abilities of the POL1 and POL2 isolates to metabolize a panel of 190 carbon sources were examined using a BIOLOG assay. The results confirmed the abilities of the shewanellas to utilize a broad range of carbon substrates. The utility of the POL1 and POL2 isolates in H-type MFCs operating on pyruvate or molasses was demonstrated. The operation of the MFC with shewanellas cultured on molasses was shown for the first time. A two-stage character of the fuel cell polarization curves, not previously noted in Shewanella MFC studies, was observed.