8B2D

CRYSTAL STRUCTURE OF BACTERIAL FLAVIN CONTAINING MONOOXYGENASE THERMORESISTANT MUTANT, IN COMPLEX WITH NADP+

8B2D の概要

• エントリーDOI: 10.2210/pdb8b2d/pdb
• 分子名称: Flavin-containing monooxygenase, Fmo, SULFATE ION, GLYCEROL, ... (8 entities in total)
• 機能のキーワード: thermoresistant mutant, complex, flavin adenine dinucleotide, nicotinamide adenine dinucleotide phosphate, oxidoreductase
• 由来する生物種: Methylophaga aminisulfidivorans MP
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 112345.33

構造登録者

Cea-Rama, I.,Sanz-Aparicio, J.,Ferrer Martinez, M.,Goris, M.,Bjerga, G. (登録日: 2022-09-13, 公開日: 2023-06-14, 最終更新日: 2024-02-07)

主引用文献

Goris, M.,Cea-Rama, I.,Puntervoll, P.,Ree, R.,Almendral, D.,Sanz-Aparicio, J.,Ferrer, M.,Bjerga, G.E.K.
Increased Thermostability of an Engineered Flavin-Containing Monooxygenase to Remediate Trimethylamine in Fish Protein Hydrolysates.
Appl.Environ.Microbiol., 89:e0039023-e0039023, 2023

PubMed Abstract: Protein hydrolysates made from marine by-products are very nutritious but frequently contain trimethylamine (TMA), which has an unattractive fish-like smell. Bacterial trimethylamine monooxygenases can oxidize TMA into the odorless trimethylamine -oxide (TMAO) and have been shown to reduce TMA levels in a salmon protein hydrolysate. To make the flavin-containing monooxygenase (FMO) Methylophaga aminisulfidivorans trimethylamine monooxygenase (mFMO) more suitable for industrial application, we engineered it using the Protein Repair One-Stop Shop (PROSS) algorithm. All seven mutant variants, containing 8 to 28 mutations, displayed increases in melting temperature of between 4.7°C and 9.0°C. The crystal structure of the most thermostable variant, mFMO_20, revealed the presence of four new stabilizing interhelical salt bridges, each involving a mutated residue. Finally, mFMO_20 significantly outperformed native mFMO in its ability to reduce TMA levels in a salmon protein hydrolysate at industrially relevant temperatures. Marine by-products are a high-quality source for peptide ingredients, but the unpleasant fishy odor caused by TMA limits their access to the food market. This problem can be mitigated by enzymatic conversion of TMA into the odorless TMAO. However, enzymes isolated from nature must be adapted to industrial requirements, such as the ability to tolerate high temperatures. This study has demonstrated that mFMO can be engineered to become more thermostable. Moreover, unlike the native enzyme, the best thermostable variant efficiently oxidized TMA in a salmon protein hydrolysate at industrial temperatures. Our results present an important next step toward the application of this novel and highly promising enzyme technology in marine biorefineries.

PubMed: 37222584
DOI: 10.1128/aem.00390-23

実験手法

X-RAY DIFFRACTION (1.62 Å)