Investigation of magnetic nanosomes as multifunctional nanomachines

• Applicant: Dr. Sebastian Kolinko
• Subject Area: Biological Process Engineering ; Microbial Ecology and Applied Microbiology ; Metabolism, Biochemistry and Genetics of Microorganisms
• Term: from 2015 to 2018
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 288781113

The naturally occurring cellulosome is one of the most elegant and elaborate macromolecules for nanoscale organization of synergistic enzymes. These self-assembling systems produced by cellulolytic bacteria consolidate multiple enzymes through specific cohesin-dockerin interactions in close proximity for the coordinated deconstruction of cellulose. In biotechnology nanoparticle-based immobilization techniques are used for nanoscale organization, leading to increased enzyme efficiency, stability, and enables new and fascinating applications through the combination of nanoparticle specific physical, chemical, optical and electrical properties with biological catalysis. However, due to limitations of prevalent chemical immobilization techniques most enzyme-based biotechnological applications have been limited to single effector proteins. To enable controlled immobilization of multiple effector proteins, increase efficiency and recovery the aim of this project is to construct recyclable multifunctional nanobiocatalysts by combining magnetic nanoparticles with nanoscale immobilization of synergistic enzymatic pathways. Applying synthetic biology I will design artificial protein scaffolds for enzyme immobilization and genetically fuse them to biosynthesized membrane enveloped, nanoparticles (magnetosomes), resulting in magnetic multifunctional nanobiocatalysts. Designed scaffolds will contain different cellulosome-borne cohesin domains that will recruit desired moieties by specific interaction with their cognate dockerin domain. Furthermore, scaffolds will be genetically fused to an anchor protein within the biomembrane enveloping biosynthesized magnetosomes.The assembly and functionality of different multifunctional nanobiocatalysts will be evaluated by fluorescence microscopy, various biochemical approaches, and degradation of recalcitrant biomass. In addition, I will analyze the magnetic recyclability of functionalized magnetic nanoparticles from one-pot reactions. The proposed project will contribute to the foundational understanding of nanoscale enzyme organization and will push the frontier of functionalized magnetosomes. Ultimately, it has direct applications in biotechnology and the potential to develop highly efficient enzyme and environmentally friendly applications like biofuel production from lignocellulosic biomass.

• DFG Programme: Research Fellowships
• International Connection: USA
• Host: Steven Singer, Ph.D.