ИСТИНА

The spread of antibiotic resistance is one of the major concerns for modern medicine. One promising approach to solving the problem is the discovery of new antibacterial compounds. At the same time, traditional screening of soil actinomycetes for the production of antimicrobial agents demands much time and often yields known molecules. One possible solution to this issue is crowdsourcing. The Citizen Science project is currently being carried out in Russia, starting from 2021. In this project, we decided to give school students and the interested public a unique opportunity to collect and analyze natural samples in educational (school) laboratories which are not specially equipped, or even at home. Therefore, we have developed an optimized workflow divided into two stages: (a) isolation and screening for new producing strains; (b) purification and identification of active compounds. The first stage can be carried out in non-sterile conditions by volunteers using a special Isolation Kit, while the second stage is carried out in the research center. Two reporter plasmids encoding visible fluorescent protein genes have been created to provide primary information about the action mechanism of antimicrobial agents. In addition, activity-guided HPLC analysis followed by high-resolution mass spectrometry (HRMS) provides a comprehensive dataset on the active compound making it possible to identify the compound using various databases on natural products. As a proof of concept, two new strains, Streptomyces sp. KB-1 and BV113, were found to produce the antibiotics pikromycin and chartreusin, respectively. Thus, we have demonstrated that the novel workflow of the Citizen Science project is an effective means to improve the screening productivity and increase discovery rates. This research was funded by the Ministry of Science and Higher Education of the Russian Federation (Agreement NO. 075–15-2021-1085).