The engineered biphenyl dioxygenases enhanced the metabolism of dibenzofuran

Wang, Yuan; Sun, Chengcheng; Min, Jin‐Young; Li, Bingjun; Li, Junde; Chen, Weiwei; Kong, Yachao; Hu, Xiaoke

doi:10.1016/j.ibiod.2021.105228

Cited by 4 publications

(4 citation statements)

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“…This results in the substrate scopes and the activities of these enzymes being restricted by the size and by the electronics of any potential substrates (10,35). To alleviate these restrictions on the utility of Rieske dioxygenases, multiple research groups have applied the tools of enzyme engineering to improve their reactivity or to expand their substrate scopes (5,3,11,20,26,33,37,38). This has included studies that have engineered Rieske dioxygenases specifically to improve their utility in the bioremediation of aromatic pollutants in the soil (1,21).…”

Section: Figurementioning

confidence: 99%

“…Our lab has recently reported the development of improved Rieske dioxygenase variants through the application of rational enzyme engineering (27). These studies have led to the development of Rieske dioxygenases with improved reactivity or expanded substrate scopes, increasing the practical utility of these green chemical tools (1,3,5,11,20,21,26,27,33,37,38).…”

Section: Figurementioning

confidence: 99%

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Investigation of the Effects of Mutating Iron-Coordinating Residues in Rieske Dioxygenases

Froese,

Betts

2024

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Rieske dioxygenases are multi-component enzyme systems, naturally found in many soil bacteria, that have been widely applied in the production of fine chemicals, owing to the unique and valuable oxidative dearomatization reactions they catalyze. The range of practical applications for these enzymes in this context has historically been limited, however, due to their limited substrate scope and strict selectivity. In an attempt to overcome these limitations, our research group has employed the tools of enzyme engineering to expand the substrate scope or improve the reactivity of these enzyme systems in specific contexts. Traditionally, enzyme engineering campaigns targeting metalloenzymes have avoided mutations to metal-coordinating residues, based on the assumption that these residues are essential for enzyme activity. Inspired by the success of other recent enzyme engineering reports, our research group investigated the potential to alter or improve the reactivity of Rieske dioxygenases by altering or eliminating iron coordination in the active site of these enzymes. Herein, we report the modification of all three iron-coordinating residues in the active site of toluene dioxygenase both to alternate residues capable of coordinating iron, and to a residue that would eliminate iron coordination. The enzyme variants produced in this way were tested for their activity in the cis-dihydroxylation of a small library of potential aromatic substrates. The results of these studies demonstrated that all three iron-coordinating residues, in their natural state, are essential for enzyme activity in toluene dioxygenase, as the introduction of any mutations at these sites resulted in a complete loss of cis-dihydroxylation activity.

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Section: Figurementioning

confidence: 99%

Section: Figurementioning

confidence: 99%

Investigation of the Effects of Mutating Iron-Coordinating Residues in Rieske Dioxygenases

Froese,

Betts

2024

finefocus

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“…Despite the catalytic versatility demonstrated by RDOs, the substrate scope and activity of these enzymes remain limited by the steric and electrostatic demands of potential substrates. 7–9 As a result of these restrictions on the activity and selectivity of the native enzymes, the range of synthetic applications for RDOs has remained limited. To alleviate these constraints on the range of applications for RDOs, enzyme engineering has been applied to expand their substrate scopes and to improve the activity of these enzymes for specific classes of substrates.…”

mentioning

confidence: 99%

“…To alleviate these constraints on the range of applications for RDOs, enzyme engineering has been applied to expand their substrate scopes and to improve the activity of these enzymes for specific classes of substrates. 9 These studies have been successful both in improving the activity of RDOs for specific substrate classes, making their application in organic synthesis more practical, 9 d,e , m , t , x ,10 and in developing RDO variants with activity for entirely new substrates. 9 h , n , o , q…”

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confidence: 99%