Large‐scale culturing of D. mccartyi strain NIT01

The large‐scale culturing of D. mccartyi strain NIT01 was performed using three different types of vessels: an 11.4 L‐SUS304 vessel (FUJITECHNO Co., Ltd., Tokyo, Japan), a 10 L‐glass vessel (FB‐800‐10000, Wexer Lifescience Co., Ltd., Tokyo, Japan), and an 18 L‐titanium vessel (Saito Industry Co., Ltd., Aichi, Japan), wherein the working volumes were 7, 7, and 10 L, respectively. First, the DHB‐CO₃‐Br medium was autoclaved in closed vessels made of SUS304 and titanium. After covering the glass vessel with aluminium foil, its medium was autoclaved before bubbling the media with the N₂‐CO₂ gas mixture to avoid breakage of the vessel. The headspace was then replaced with the H₂‐CO₂ gas mixture gas and supplemented with acetate, reducing agents, and vitamins. To inoculate D. mccartyi strain NIT01, 0.5–1.6 L of the pre‐culture was first placed in a glass vial and connected to the closed vessel via a needle and a silicon tube, and then injected into the large‐scale vessels by adding the H₂‐CO₂ gas mixture to the glass vial. The inoculum rate for culturing in the glass and SUS304 vessels was 19%, and that in the titanium vessel was 5% or 16% (v/v) (Figure S1). Finally, TCE was added at a concentration of 0.24–3.0 mM to the vessel. D. mccartyi strain NIT01 was cultured using the large‐scale vessels at least four times.