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.
journal_name
Nat Communjournal_title
Nature communicationsauthors
Yeom SJ,Kim M,Kwon KK,Fu Y,Rha E,Park SH,Lee H,Kim H,Lee DH,Kim DM,Lee SGdoi
10.1038/s41467-018-07488-0subject
Has Abstractpub_date
2018-11-29 00:00:00pages
5053issue
1issn
2041-1723pii
10.1038/s41467-018-07488-0journal_volume
9pub_type
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