Plasmids were transferred from E. c

Plasmids were transferred from E. coli S17-1 to recombinant H. bluephagenesis TD1.0 via conjugation. Briefly, single colonies of E. coli S17-1 harboring plasmids of interest (donor cell) and recombinant H. bluephagenesis TD1.0 (recipient cell) were grown overnight in LB medium and 60 LB medium, respectively, in the presence of relevant antibiotic(s). Subsequently, overnight cultures were inoculated into a fresh medium at 1% volume until its OD600 reached 0.8. Cells were harvested via centrifugation at 1500×g for 2 min (ThermoFisher Scientific, Sorvall Legend Micro21R, USA), followed by washing twice with fresh LB and 60 LB medium, respectively. Following this, donor and recipient cells were mixed at a ratio of 1:1 into 50 μL of a 60LB medium, then spread on 20 LB agar plates for overnight incubation at 37 ◦C. Finally, a single colony was picked and re-suspended in 50 μL of 60 LB medium. Next, 50 μL cultures were spread on 60LB agar plates in the presence of the relevant antibiotics for 24–48 h growth at 37 ◦C. Positive colonies were selected for further studies.