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Journal of Biomedicine and Biotechnology
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type: Information Content Entity
publication venue for: Engineering and directed evolution of a Ca2 binding site A-Deficient AprE mutant reveal an essential contribution of the loop leu 75 -leu 82 to enzyme activity
type: Continuant
type: Entity
type: Generically Dependent Continuant
type: Document
type: Collection
type: Periodical
type: Journal

individual
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date: 2026-08-28T02:00:09
publisher: vivo
type: Document
label: RDF description of Engineering and directed evolution of a Ca2 binding site A-Deficient AprE mutant reveal an essential contribution of the loop leu 75 -leu 82 to enzyme activity - http://orbis.uaslp.mx/vivo/individual/scopus2337

Article
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Information Content Entity
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Engineering and directed evolution of a Ca2 binding site A-Deficient AprE mutant reveal an essential contribution of the loop leu 75 -leu 82 to enzyme activity
type: Document
type: Entity
estado: published
abstract: An aprE mutant from B. subtilis 168 lacking the connecting loop Leu 75 - Leu 82 which is predicted to encode a Ca 2 binding site was constructed. Expression of the mutant gene (aprE Δ L e u 75 - L e u 82) produced B. subtilis colonies lacking protease activity. Intrinsic fluorescence analysis revealed spectral differences between wild-type AprE and AprE Δ L 75 - L 82. An AprE Δ L 75 - L 82 variant with reestablished enzyme activity was selected by directed evolution. The novel mutations Thr 66 Met/ Gly 102 Asp located in positions which are predicted to be important for catalytic activity were identified in this variant. Although these mutations restored hydrolysis, they had no effect with respect to thermal inactivation of AprE Δ L 75 - L 82 T 66 M G 102 D. These results support the proposal that in addition to function as a calcium binding site, the loop that connects -sheet e3 with -helix c plays a structural role on enzyme activity of AprE from B. subtilis 168. Copyright © 2009 Eliel R. Romero-Garćia et al.
related by: authorship_13856
pmid: 19710937
e iscopus: 2-s2.0-69249213494
freetext keyword: bacterial protein; calcium ion; glycine; leucine; methionine; protein apre; threonine; unclassified drug; AprE protein, Bacteria; calcium; carrier protein; article; Bacillus subtilis; bacterial strain; bacterium colony; binding site; calcium binding; catalysis; controlled study; enzyme activity; fluorescence analysis; gene expression; nonhuman; protein hydrolysis; chemical structure; chemistry; directed molecular evolution; enzyme stability; enzymology; genetics; kinetics; metabolism; methodology; protein folding; site directed mutagenesis; spectrofluorometry; structure activity relation; Bacillus subtilis; Bacterial Proteins; Binding Sites; Calcium; Directed Molecular Evolution; Enzyme Stability; Kinetics; Membrane Transport Proteins; Models, Molecular; Mutagenesis, Site-Directed; Protein Folding; Spectrometry, Fluorescence; Structure-Activity Relationship
related by: authorship_13860
crisid: 2338
volume: 2009
related by: authorship_13854
type: Information Content Entity
related by: authorship_13859
doi: 10.1155/2009/201075
type: Article
date time value: article date 2338
related by: authorship_13861
abstract: An aprE mutant from B. subtilis 168 lacking the connecting loop Leu 75 - Leu 82 which is predicted to encode a Ca 2 %2b binding site was constructed. Expression of the mutant gene (aprE Δ L e u 75 - L e u 82) produced B. subtilis colonies lacking protease activity. Intrinsic fluorescence analysis revealed spectral differences between wild-type AprE and AprE Δ L 75 - L 82. An AprE Δ L 75 - L 82 variant with reestablished enzyme activity was selected by directed evolution. The novel mutations Thr 66 Met/ Gly 102 Asp located in positions which are predicted to be important for catalytic activity were identified in this variant. Although these mutations restored hydrolysis, they had no effect with respect to thermal inactivation of AprE Δ L 75 - L 82 T 66 M G 102 D. These results support the proposal that in addition to function as a calcium binding site, the loop that connects -sheet e3 with -helix c plays a structural role on enzyme activity of AprE from B. subtilis 168. Copyright © 2009 Eliel R. Romero-Garćia et al.
related by: authorship_13857
accesstype: All Open Access, Gold, Green
artnum: 201075
type: Thing
type: Generically Dependent Continuant
label: Engineering and directed evolution of a Ca2%2b binding site A-Deficient AprE mutant reveal an essential contribution of the loop leu 75 -leu 82 to enzyme activity
chemicalscas: calcium ion, 14127-61-8; glycine, 56-40-6, 6000-43-7, 6000-44-8; leucine, 61-90-5, 7005-03-0; methionine, 59-51-8, 63-68-3, 7005-18-7; threonine, 36676-50-3, 72-19-5; calcium, 7440-70-2; carrier protein, 80700-39-6; AprE protein, Bacteria; Bacterial Proteins; Calcium, 7440-70-2; Membrane Transport Proteins
related by: authorship_13855
most specific type: Document
has publication venue: Journal of Biomedicine and Biotechnology
origen: scopus
most specific type: Article
type: Continuant
related by: authorship_13858