Literature summary for 3.5.1.24 extracted from

Chand, D.; Panigrahi, P.; Varshney, N.; Ramasamy, S.; Suresh, C.G.
Structure and function of a highly active bile salt hydrolase (BSH) from Enterococcus faecalis and post-translational processing of BSH enzymes (2018), Biochim. Biophys. Acta, 1866, 507-518 .

Cloned(Commentary)

Cloned (Comment)	Organism
gene bsh, sequence comparisons, recombinant expression of wild-type and mutant C-terminally His-tagged enzyme in Escherichia coli strain BL21	Enterococcus faecalis

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant His-tagged wild-type enzyme and mutant C2A, hanging drop vapour diffusion method, mixing of 0.002 ml of 15-20 mg/ml protein solution and reservoir solution each, the latter containing 2 M ammonium sulfate and 5% isopropyl alcohol, 2-3 days, X-ray diffraction structure determination and analysis at 2.0 and 1.5 A resolution, respectively, molecular replacement using the crystal structure of BSH from Clostridium perfringenes (PDB ID 2RLC, chain A) as a template for the search model	Enterococcus faecalis

Crystallization (Comment)

Organism

purified recombinant His-tagged wild-type enzyme and mutant C2A, hanging drop vapour diffusion method, mixing of 0.002 ml of 15-20 mg/ml protein solution and reservoir solution each, the latter containing 2 M ammonium sulfate and 5% isopropyl alcohol, 2-3 days, X-ray diffraction structure determination and analysis at 2.0 and 1.5 A resolution, respectively, molecular replacement using the crystal structure of BSH from Clostridium perfringenes (PDB ID 2RLC, chain A) as a template for the search model

Enterococcus faecalis

Protein Variants

Protein Variants	Comment	Organism
N79W	site-directed mutagenesis, the mutant shows a drastic decrease in both expression and BSH activity and no penicillin V acylase activity compared to the wild-type enzyme	Enterococcus faecalis
N79Y	site-directed mutagenesis, the mutant shows good expression retaining about 88% wild-type BSH activity but has no penicillin V acylase activity	Enterococcus faecalis
Y20W/N79W	site-directed mutagenesis, the mutant shows a drastic decrease in both expression and BSH activity and no penicillin V acylase activity compared to the wild-type enzyme	Enterococcus faecalis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
glycocholic acid + H2O	Enterococcus faecalis	-	cholate + glycine	-	?

Organism

Organism	UniProt	Comment	Textmining
Enterococcus faecalis	Q83YZ2	-	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
proteolytic modification	the enzyme performs autocatalytic mechanism, overview	Enterococcus faecalis

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged wild-type and mutant enzyme from Escherichia coli strain BL21 by nickel affinity chromatography and gel filtration to apparent homogeneity	Enterococcus faecalis

Source Tissue

Source Tissue	Comment	Organism	Textmining

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
glycocholic acid + H2O	-	Enterococcus faecalis	cholate + glycine	-	?
additional information	the enzyme preforms autocatalytic processing, mechanism, overview. The wild-type enzyme also exhibits penicillin V acylase activity	Enterococcus faecalis	?	-	?

Subunits

Subunits	Comment	Organism
More	the structural analysis reveals a reduced secondary structure content compared to other known BSH structures, particularly devoid of an anti-parallel beta-sheet in the assembly loop and part of a beta-strand is converted to increase the length of a substrate binding loop 2. Three-dimensional structure analysis, overview	Enterococcus faecalis
tetramer	-	Enterococcus faecalis

Synonyms

Synonyms	Comment	Organism
BSH	-	Enterococcus faecalis
EfBSH	-	Enterococcus faecalis
salt hydrolase	-	Enterococcus faecalis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
50	-	assay at	Enterococcus faecalis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Enterococcus faecalis

General Information

General Information	Comment	Organism
evolution	bile salt hydrolase (BSH), a member of cholylglycine hydrolase family, catalyzes the de-conjugation of bile acids and is evolutionarily related to penicillin V acylase (PVA) that hydrolyses a different substrate such as penicillin V. Enterococcus faecalis bile salt hydrolase (BSH) has a manifold higher hydrolase activity compared to other known BSHs and displays unique allosteric catalytic property. The structural analysis reveals a reduced secondary structure content compared to other known BSH structures, particularly devoid of an anti-parallel beta-sheet in the assembly loop and part of a beta-strand is converted to increase the length of a substrate binding loop 2. An evolution-mediated subordination of the pre-peptide excision site results in loss of pre-peptide in EfBSH and other related cholylglycine hydrolases	Enterococcus faecalis
malfunction	two serine residues next to nucleophile cysteine are essential for autocalytic processing to remove precursor peptide, but since a pre-peptide is absent in EfBSH the mutation of these serines is tolerated	Enterococcus faecalis
additional information	the structural analysis reveals a reduced secondary structure content compared to other known BSH structures, particularly devoid of an anti-parallel beta-sheet in the assembly loop and part of a beta-strand is converted to increase the length of a substrate binding loop 2. The substrate binding pocket shows reduced volume owing to altered loop conformations and increased hydrophobicity contributed by a higher ratio of hydrophobic to hydrophilic groups present. The aromatic residues F18, Y20 and F65 participate in substrate binding. Effective polar complementarity present for the three hydroxyl groups of glycocholic acid substrate in the binding site contributing to higher substrate specificity and efficient catalysis. These are unique features characteristics of this BSH enzyme and thought to contribute to its higher activity and specificity towards bile salts as well as allosteric effects. Substrate docking and molecular dynamics simulation studies, overview	Enterococcus faecalis