A synthetic microbial biosensor for high-throughput screening of lactam biocatalysts

Soo-Jin Yeom, Moonjeong Kim, Kil Koang Kwon, Yaoyao Fu, Eugene Rha, Sung-Hyun Park, Hyewon Lee, Haseong Kim, Dae-Hee Lee, Dong-Myung Kim and Seung-Goo Lee ()
Additional contact information
Soo-Jin Yeom: KRIBB
Moonjeong Kim: KRIBB
Kil Koang Kwon: KRIBB
Yaoyao Fu: KRIBB
Eugene Rha: KRIBB
Sung-Hyun Park: KRIBB
Hyewon Lee: KRIBB
Haseong Kim: KRIBB
Dae-Hee Lee: KRIBB
Dong-Myung Kim: Chungnam National University
Seung-Goo Lee: KRIBB

Nature Communications, 2018, vol. 9, issue 1, 1-12

Abstract: Abstract Biocatalytic cyclization is highly desirable for efficient synthesis of biologically derived chemical substances, such as the commodity chemicals ε-caprolactam and δ-valerolactam. To identify biocatalysts in lactam biosynthesis, we develop a caprolactam-detecting genetic enzyme screening system (CL-GESS). The Alcaligenes faecalis regulatory protein NitR is adopted for the highly specific detection of lactam compounds against lactam biosynthetic intermediates. We further systematically optimize the genetic components of the CL-GESS to enhance sensitivity, achieving 10-fold improvement. Using this highly sensitive GESS, we screen marine metagenomes and find an enzyme that cyclizes ω-amino fatty acids to lactam. Moreover, we determine the X-ray crystal structure and catalytic residues based on mutational analysis of the cyclase. The cyclase is also used as a helper enzyme to sense intracellular ω-amino fatty acids. We expect this simple and accurate biosensor to have wide-ranging applications in rapid screening of new lactam-synthesizing enzymes and metabolic engineering for lactam bio-production.

Date: 2018
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DOI: 10.1038/s41467-018-07488-0

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