back

Spores with pFiF

Spores with pFiF

Summary of the idea behind spores with pFiF. (1) DNA sequence is selected as ID and is protected by B. Subtilis in spores. (2) Spore-protected, robust DNA is integrated into printing inks. (3) Industrial 2D and 3D printing processes are used to

Period: 1.1.2020 – 31.12.2021

---

Prof. Dr. Johannes Kabisch |

Dept. Biology| computational synthetic biology

Dr.-Ing. Dieter Spiehl |

Dept. Mechanical Engineering | Institute for Printing Machines and Printing Processes

---

Project description:

Part labeling is a crucial security feature as it can prevent product counterfeiting. DNA, the information carrier of life, is started to be explored as an engineered information molecule with immense potential in respect to information density and encryption potential. Most research in this direction is concerned with how to encode binary data to DNA and read the stored information from this DNA. Little to no effort is made on how to apply DNA and the information stored within as an identification label for security. In this study, we explore DNA in various printing processes for its suitability as an anti‐counterfeiting and identification tag. DNA is sensitive to environmental influences, which is why we compare the suitability of free DNA against using the spores of the bacterium Bacillus subtilis as a naturally evolved DNA protective shell and a cheap way for mass production of DNA. To integrate these two different variants into products, we use both conventional printing methods and additive manufacturing processes. We investigate the stresses on the spores, derive suitable printing techniques and assess the practical application ‐ processing, extraction and subsequent detection via PCR ‐ using selected printing methods. The stresses are differentiated into four groups ‐ solvents, UV irradiation, temperature and shear stress. with a significant advantage of DNA protected in spores only for selected solvents. However, in actual printing processes stresses are combined and thus we test two exemplary and complementary methods. Namely gravure printing as a 2D and masked stereolithography as a 3D printing method. The combination of even low stresses in gravure printing of water-based inks makes it impossible to detect free DNA via PCR whereas a detection of the DNA protected in spores is possible. In addition, a solvent based gravure printing ink is tested and the selected 3D printing method produces a solid part out of UV-curable resin. From all three examples, the extraction of a sample and subsequently the detection of the DNA from the spores could be archieved. Hence, we were able to show that the production of a robust DNA‐based counterfeit protection and product integration is possible with industrial 2D and 3D printing processes.

Project website

· Dörsam, E., Euler, T., Fergen, I., Haas, M., Kurmakaev, E., Schmitt-Lewen, M., & Sonnenschein, J. (2012). Herstellung eines Merkmals für die Fälschungssicherheit. DE102012010482A1, Prioritätsdatum 18.06.2011, anhängig

· Fernandes, F., Schmitt-Lewen, M., & Dörsam, E. (2019). Herstellung von Sicherheitskennzeichen. DE102018219252A1, Prioritätsdatum 09.04.2018, anhängig

· Jaeger, J., Groher, F., Stamm, J., Spiehl, D., Braun, J., Dörsam, E., & Suess, B. (2019). Characterization and inkjet printing of an RNA aptamer for paper-based biosensing of ciprofloxacin. Biosensors, 9(1), 7. https://doi.org/10.3390/bios9010007

· Kabisch, J., & Festel G. (2017). Oligonucleotides in food, beverage, cosmetic and medicinal formulations, WO2017092808A1, Priority date 12.03.2015, pending

· Kabisch, J., Schlichting, N., & Karava, M. (unveröffentlicht). Rapid transformation method for Gram-positive bacteria. EP20187230, Priority date 22.06.2020, pending

· Kabisch, J., Thürmer, A., Hübel, T., Popper, L., Daniel, R., & Schweder, T. (2013). Characterization and optimization of Bacillus subtilis ATCC 6051 as an expression host. Journal of biotechnology, 163(2), 97-104. https://doi.org/10.1016/j.jbiotec.2012.06.034

· Karava, M., Bracharz, F., & Kabisch, J. (2019). Quantification and Isolation of Bacillus subtilis Spores using Cell Sorting and automated Gating. PloS one, 14(7), e0219892. https://doi.org/10.1371/journal.pone.0219892

· Karava, M., Gockel, P., & Kabisch J. (2020). Bacillus subtilis spore surface display of photodecarboxylase for the transformation of lipids to hydrocarbons. bioRxiv. https://doi.org/10.1101/2020.08.30.273821

· Kumpfmueller, J., Kabisch, J., & Schweder, T. (2013). An optimized technique for rapid genome modifications of Bacillus subtilis. Journal of Microbiological Methods, 95(3), 350-352. https://doi.org/10.1016/j.mimet.2013.10.003

· Nadler, F., Bracharz, F., & Kabisch, J., (2019). CopySwitch – in vivo Optimization of Gene Copy Numbers for Heterologous Gene Expression in Bacillus subtilis. Front. Bioeng. Biotechnol., 6, 207. https://doi.org/10.3389/fbioe.2018.00207

· Stamm, J., Spiehl, D., Jaeger, J., Groher, F., Meckel, T., Suess, B., & Dörsam, E. (2019). A test system for the printing of functional nucleic acids onto different carriers and verification of its functionality by DNA dyes. Journal of Print and Media Technology Research, 8(1), 7-17. https://doi.org/10.14622/JPMTR-1821