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High-throughput Screening and Selection of Artificial Lactam Riboswitch
Junyeong SUNG,Sungyeon JANG,Sungho JANG,Dae-Kyun IM,Teak jun KANG,Min-Kyu OH,Gyoo Yeol JUNG 한국생물공학회 2021 한국생물공학회 학술대회 Vol.2021 No.10
Caprolactam is a monomer used for the synthesis of nylon-6. An intracellular biosensor for caprolactam can facilitate high-throughput metabolic engineering of recombinant microbial strains. Because of the mixed production of caprolactam and valerolactam in the recombinant strain, a caprolactam biosensor should be highly specific for caprolactam. However, a highly specific caprolactam sensor has not been reported. Here, we developed an artificial riboswitch that specifically responds to caprolactam. This riboswitch was prepared using a coupled in vitro-in vivo selection strategy with a heterogeneous pool of RNA aptamers obtained from in vitro selection to construct a riboswitch library used in in vivo selection. The caprolactam riboswitch successfully discriminated caprolactam from valerolactam. This riboswitch enabled caprolactam-dependent control of cell growth, which will be useful for improving caprolactam production and is a valuable tool for metabolic engineering.