Journal: Applied Microbiology and Biotechnology

Article Title: Enhancing the imidase activity of BpIH toward 3-isobutyl glutarimide via semi-rational design

doi: 10.1007/s00253-024-13311-2

Figure Lengend Snippet: Binding diagram of IBI with BpIH. a Overall structure of the homotetramer BpIH. b Four spatial positions of IBI in BpIH subunit A according to the docking results. c Interaction of IBI with BpIH in position 1. d Interaction of IBI with BpIH in position 2. e Interaction of IBI with BpIH in position 3. f Interaction of IBI with BpIH in position 4. The substrate IBI is shown as sticks

Article Snippet: The sequences of the imidase BpIH and allantoinase PuuE family proteins mentioned in this study are available from the National Centre for Biotechnology Information (NCBI).

Techniques: Binding Assay

Journal: Applied Microbiology and Biotechnology

Article Title: Enhancing the imidase activity of BpIH toward 3-isobutyl glutarimide via semi-rational design

doi: 10.1007/s00253-024-13311-2

Figure Lengend Snippet: Enzyme activity determination of WT BpIH and mutants. a Alanine scanning of 17 residues within 5 Ȧ of the substrate (IBI). b Semi-saturated mutagenesis of G167 and W131

Article Snippet: The sequences of the imidase BpIH and allantoinase PuuE family proteins mentioned in this study are available from the National Centre for Biotechnology Information (NCBI).

Techniques: Activity Assay, Mutagenesis

Journal: Applied Microbiology and Biotechnology

Article Title: Enhancing the imidase activity of BpIH toward 3-isobutyl glutarimide via semi-rational design

doi: 10.1007/s00253-024-13311-2

Figure Lengend Snippet: Conservation analysis of residues within 9 Ȧ and site-directed mutagenesis of residues within 6–9 Ȧ of the binding pocket. a Structure superimposition of WT BpIH and the allantoinase family proteins. b Conservativity analysis of the residues within 5 Ȧ (upper) and 6–9 Ȧ (lower) of the binding pocket. The letter size represents the degree of conservation of the residue. c Relative activity of WT BpIH and mutants of residues within 6–9 Ȧ of the binding pocket. d Relative activity of WT BpIH and mutants of S226

Article Snippet: The sequences of the imidase BpIH and allantoinase PuuE family proteins mentioned in this study are available from the National Centre for Biotechnology Information (NCBI).

Techniques: Mutagenesis, Binding Assay, Residue, Activity Assay

Journal: Applied Microbiology and Biotechnology

Article Title: Enhancing the imidase activity of BpIH toward 3-isobutyl glutarimide via semi-rational design

doi: 10.1007/s00253-024-13311-2

Figure Lengend Snippet: Comparison of the activity of WT BpIH and the combinatorial mutants of residues Y37, H133, and S226. The relative specific enzyme activity of YHS-I is shown in lime green, and the remaining mutant strains are shown in green. YHS-V, Y37FH133NS226V; YHS-I, Y37FH133NS226I; YHS-L, Y37FH133NS226L

Article Snippet: The sequences of the imidase BpIH and allantoinase PuuE family proteins mentioned in this study are available from the National Centre for Biotechnology Information (NCBI).

Techniques: Comparison, Activity Assay, Mutagenesis

Journal: Applied Microbiology and Biotechnology

Article Title: Enhancing the imidase activity of BpIH toward 3-isobutyl glutarimide via semi-rational design

doi: 10.1007/s00253-024-13311-2

Figure Lengend Snippet: Kinetic parameters and enantiopurity of WT BpIH and its mutants

Article Snippet: The sequences of the imidase BpIH and allantoinase PuuE family proteins mentioned in this study are available from the National Centre for Biotechnology Information (NCBI).

Techniques:

Journal: Applied Microbiology and Biotechnology

Article Title: Enhancing the imidase activity of BpIH toward 3-isobutyl glutarimide via semi-rational design

doi: 10.1007/s00253-024-13311-2

Figure Lengend Snippet: R -IBM conversion from IBI catalyzed by WT BpIH and three mutants (S226L, YHS-L, and YHS-I)

Article Snippet: The sequences of the imidase BpIH and allantoinase PuuE family proteins mentioned in this study are available from the National Centre for Biotechnology Information (NCBI).

Techniques:

Journal: Applied Microbiology and Biotechnology

Article Title: Enhancing the imidase activity of BpIH toward 3-isobutyl glutarimide via semi-rational design

doi: 10.1007/s00253-024-13311-2

Figure Lengend Snippet: Molecular dynamics study of WT BpIH and the YHS-I mutant. MD simulations were performed over a 40-ns time period at 40 °C. a RMSD. b RMSF. c Stability at 40 °C. Each experiment was repeated three times. d Overlay of the average structures extracted from the final 10 ns of the equilibrated state of the MD simulations, with proteins shown in cartoon form (white) and mutant site residues shown in stick form (green)

Article Snippet: The sequences of the imidase BpIH and allantoinase PuuE family proteins mentioned in this study are available from the National Centre for Biotechnology Information (NCBI).

Techniques: Mutagenesis