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. 2020 Sep 18;11:578533. doi: 10.3389/fmicb.2020.578533

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Copyright © 2020 Ayoola, Nakamya, Shack, Park, Lim, Lee, Ross, Eoh and Nanduri.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Agmatine is critical for capsule biosynthesis in pneumococcus. Total CPS isolated from ΔspeA and ΔpotABCD cultured in THY or THY supplemented with agmatine [20 mM of agmatine, a quarter of minimum inhibitory concentration (MIC)] was spotted on a nitrocellulose membrane. Probing with anti-serotype 4 specific antibody and a horseradish peroxidase (HRP)-conjugated secondary antibody, the membrane was developed with enhanced chemiluminescence (ECL) detection and scanned using a ChemiDoc XRS+ with Image Lab software (Bio-Rad, Hercules, CA, United States). Agmatine successfully restored capsule in the two unencapsulated ΔspeA and ΔpotABCD strains in three independent experiments.