5YSS

Crystal structure of aminocaproic acid cyclase in complex with NAD (+)

5YSS の概要

• エントリーDOI: 10.2210/pdb5yss/pdb
• 分子名称: 3-hydroxybutyrate dehydrogenase, NICOTINAMIDE-ADENINE-DINUCLEOTIDE (3 entities in total)
• 機能のキーワード: cyclase, nad binding protein, complex, oxidoreductase
• 由来する生物種: Citrobacter freundii
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 109329.55

構造登録者

Fu, Y.,Yeom, S.J.,Lee, S.G. (登録日: 2017-11-15, 公開日: 2018-10-31, 最終更新日: 2023-11-22)

主引用文献

Yeom, S.J.,Kim, M.,Kwon, K.K.,Fu, Y.,Rha, E.,Park, S.H.,Lee, H.,Kim, H.,Lee, D.H.,Kim, D.M.,Lee, S.G.
A synthetic microbial biosensor for high-throughput screening of lactam biocatalysts
Nat Commun, 9:5053-5053, 2018

PubMed 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.

PubMed: 30498220
DOI: 10.1038/s41467-018-07488-0

実験手法

X-RAY DIFFRACTION (2.276 Å)