Site-directed mutagenesis: role of lid region for T1 lipase specificity

Mohamed, Rauda A.; Salleh, Abu Bakar; Leow, Thean Chor; Normi, Yahaya M.; Rahman, Mohd Basyaruddin Abdul

doi:10.1093/protein/gzy023

Cited by 7 publications

(2 citation statements)

References 19 publications

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“…Ramachandran plot ( Figure S3 ) displays 90.9% of the residues that reside in the most favored regions, which indicates the conformational rationality of the modeling product. EstJ6 presents a typical α/β hydrolase folding structure and possesses typical cap and lid regions, which are vital for substrate selectivity and binding [ 42 ]. The binding pocket is mainly formed by hydrophobic residues ( Figure S4 ), which is beneficial for the binding of PAEs, which are hydrophobic molecules.…”

Section: Resultsmentioning

confidence: 99%

Computational Insights into the Allosteric Modulation of a Phthalate-Degrading Hydrolase by Distal Mutations

Bao

et al. 2023

Biomolecules

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Phthalate esters (PAEs) are a ubiquitous kind of environmental endocrine that disrupt chemicals, causing environmental and health issues. EstJ6 is an effective phthalate-degrading hydrolase, and its mutant with a combination of three non-conservative distal mutations has an improved activity against PAEs with unknown molecular mechanisms. Herein, we attempt to fill the significant gap between distal mutations and the activity of this enzyme using computational approaches. We found that mutations resulted in a redistribution of the enzyme’s preexisting conformational states and dynamic changes of key functional regions, especially the lid over the active site. The outward motion of the lid upon the mutations made it easier for substrates or products to enter or exit. Additionally, a stronger substrate binding affinity and conformational rearrangements of catalytic reaction-associated residues in the mutant, accompanied by the strengthened communication within the protein, could synergistically contribute to the elevated catalytic efficiency. Finally, an attempt was made to improve the thermostability of EstJ6 upon introducing a distal disulfide bond between residues A23 and A29, and the simulation results were as expected. Together, our work explored the allosteric effects caused by distal mutations, which could provide insights into the rational design of esterases for industrial applications in the future.

show abstract

Section: Resultsmentioning

confidence: 99%

Computational Insights into the Allosteric Modulation of a Phthalate-Degrading Hydrolase by Distal Mutations

Bao

et al. 2023

Biomolecules

View full text Add to dashboard Cite

show abstract

“…Ramachandran plot (Figure S3) displays that 90.9% residues reside in the most favored regions, which indicates the conformational rationality of the modeling product. EstJ6 presents a typical α/β hydrolase folding structure, and possesses of typical cap and lid regions, which are vital for substrate selectivity and binding [42]. The binding pocket is mainly formed by hydrophobic residues (Figure S4), which is beneficial for the binding of PAEs, which are hydrophobic molecules.…”

Section: Sequential and Structural Analyses Of Estj6mentioning

confidence: 99%

Computational Insights into the Allosteric Modulation of a Phthalate-Degrading Hydrolase by Distal Mutations

Xu¹,

Bao²,

Li³

et al. 2023

Preprint

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Phthalate esters (PAEs) are a ubiquitous kind of environmental endocrine-disrupting chemicals, causing environmental and health issues. EstJ6 is an effective phthalate-degrading hydrolase, and its mutant with a combination of three non-conservative distal mutations has an improved activity against PAEs with unknown molecular mechanisms. Herein, we attempt to fill the significant gap between distal mutations and the activity of this enzyme using computational approaches. We find mutations result in a redistribution of enzyme’s preexisting conformational states and dynamics changes of key functional regions, especially the lid over the active site. The outward motion of the lid upon mutations should make it easier for substrates or products to enter or exit. Additionally, the stronger substrate binding affinity and conformational rearrangements of catalytic reaction-associated residues in mutant, accompanied by the strengthened communication within the protein, might contribute to the elevated catalytic efficiency. Finally, an attempt has been done to improve the thermostability of EstJ6 upon introducing a distal disulfide bond between residues A23 and A29, and the simulation results are as expected. Together, our work explored the allosteric effects caused by distal mutations, which could provide insights into the rational design of esterases for industrial applications in the future.

show abstract

Immobilization of lipases via interfacial activation on hydrophobic supports: Production of biocatalysts libraries by altering the immobilization conditions

et al. 2021

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Site-directed mutagenesis: role of lid region for T1 lipase specificity

Cited by 7 publications

References 19 publications

Computational Insights into the Allosteric Modulation of a Phthalate-Degrading Hydrolase by Distal Mutations

Computational Insights into the Allosteric Modulation of a Phthalate-Degrading Hydrolase by Distal Mutations

Computational Insights into the Allosteric Modulation of a Phthalate-Degrading Hydrolase by Distal Mutations

Immobilization of lipases via interfacial activation on hydrophobic supports: Production of biocatalysts libraries by altering the immobilization conditions

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