3.5.1.24: choloylglycine hydrolase
This is an abbreviated version!
For detailed information about choloylglycine hydrolase, go to the full flat file.
Word Map on EC 3.5.1.24
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3.5.1.24
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lactobacillus
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cholesterol
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deconjugation
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plantarum
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microbiota
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cholesterol-lowering
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bifidobacterium
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taurodeoxycholic
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cholic
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taurocholic
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glycocholic
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drug development
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glycodeoxycholic
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farnesoid
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co-aggregation
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pentosaceus
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rhamnosus
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glycine-conjugated
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pentosus
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pediococcus
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acylase
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fermentum
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auto-aggregation
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taurine-conjugated
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rogosa
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medicine
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food industry
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taurochenodeoxycholic
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oxgall
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nutrition
- 3.5.1.24
- lactobacillus
- cholesterol
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deconjugation
- plantarum
- microbiota
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cholesterol-lowering
- bifidobacterium
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taurodeoxycholic
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cholic
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taurocholic
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glycocholic
- drug development
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glycodeoxycholic
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farnesoid
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co-aggregation
- pentosaceus
- rhamnosus
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glycine-conjugated
- pentosus
- pediococcus
- acylase
- fermentum
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auto-aggregation
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taurine-conjugated
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rogosa
- medicine
- food industry
- taurochenodeoxycholic
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oxgall
- nutrition
Reaction
Synonyms
BAH1, BFS26_06805, bile acid hydrolase, bile salt hydrolase, Bile-Salt-Hydrolase, BSH, BSH A, BSH B, BSH C, BSH1, BSH12, BSH2, BSH3, BSH4, BSH47, BSH56, BSHA, BSHB, BSHC, BT2086, C3745_01535, CBAH, CBH, CBSHalpha, CBSHbeta, CGH, cholylglycine hydrolase, Conjugated bile acid hydrolase, conjugated bile salt hydrolase, conjugated bile salt hydrolase alpha peptide, conjugated bile salt hydrolase beta, EFBG_01849, EfBSH, glycocholase, LaciP, LgBSH, linear amide C-N hydrolase, LJ1412, LJ_0056, LJ_1147, LsalN1, lsBSH, More, probiotic bile salt hydrolase, salt hydrolase
ECTree
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Engineering
Engineering on EC 3.5.1.24 - choloylglycine hydrolase
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C1A
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site-directed mutagenesis, residue Cys1 is situated at the tip of strand beta1, inactive mutant, structure analysis in comparison to the wild-type enzyme
T2A
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site-directed mutagenesis, the mutant shows reduced activity and kcat, structure analysis in comparison to the wild-type enzyme
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
N79Y
site-directed mutagenesis, the mutant shows good expression retaining about 88% wild-type BSH activity but has no penicillin V acylase activity
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
N79V
biotechnology
Lactobacillus spp.
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enzyme activity and resistance to toxicity of bile salts are unrelated, but enzyme activity is important for Lactobacillus colonization of human intestine
C2S
site-directed mutagenesis, almost inactive mutant that shows very low remaining actiivty only with taurocholic acid
E270A
F65A
site-directed mutagenesis, the mutant shows reduced activity and altered substrate specificity compared to wild-type
N171A
site-directed mutagenesis, the mutant shows reduced activity and altered substrate specificity compared to wild-type
Q257A
site-directed mutagenesis, the mutant shows reduced activity and altered substrate specificity compared to wild-type
Y24F
site-directed mutagenesis, the mutant shows reduced activity and altered substrate specificity compared to wild-type
E270A
N171A
Ligilactobacillus salivarius NRRL B-30514
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site-directed mutagenesis, the mutant shows reduced activity and altered substrate specificity compared to wild-type
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Y24F
Ligilactobacillus salivarius NRRL B-30514
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site-directed mutagenesis, the mutant shows reduced activity and altered substrate specificity compared to wild-type
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additional information
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site-directed mutagenesis, the V79 mutation results in a stable enzyme, but reduces the catalytic activity and alters the substrate specificity of the enzyme
N79V
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site-directed mutagenesis, the V79 mutation results in a stable enzyme, but reduces the catalytic activity and alters the substrate specificity of the enzyme
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site-directed mutagenesis, the mutant shows increased activity and altered substrate specificity compared to wild-type
E270A
site-directed mutagenesis, the mutant shows reduced activity and altered substrate specificity compared to wild-type
Ligilactobacillus salivarius NRRL B-30514
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site-directed mutagenesis, the mutant shows increased activity and altered substrate specificity compared to wild-type
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E270A
Ligilactobacillus salivarius NRRL B-30514
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site-directed mutagenesis, the mutant shows reduced activity and altered substrate specificity compared to wild-type
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construction of deletion mutant BtDELTABtD2086, that is catalytically inactive. Using isogenic strains of wild-type and BSH-deleted Bacteroides thetaiotaomicron, the levels of the bile acid tauro-beta-muricholic acid in monocolonized gnotobiotic mice are selectively modulated. Bacteroides thetaiotaomicron BSH mutant-colonized mice display altered metabolism, including reduced weight gain and respiratory exchange ratios, as well as transcriptional changes in metabolic, circadian rhythm, and immune pathways in the gut and liver
additional information
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construction of deletion mutant BtDELTABtD2086, that is catalytically inactive. Using isogenic strains of wild-type and BSH-deleted Bacteroides thetaiotaomicron, the levels of the bile acid tauro-beta-muricholic acid in monocolonized gnotobiotic mice are selectively modulated. Bacteroides thetaiotaomicron BSH mutant-colonized mice display altered metabolism, including reduced weight gain and respiratory exchange ratios, as well as transcriptional changes in metabolic, circadian rhythm, and immune pathways in the gut and liver
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additional information
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substitution of the N-terminal Cys with Ser or Thr results in an inactive enzyme, which emphasizes the importance of the presence of cysteine. The enzyme is immobilized by micro-encapsulation showing low acid resistance
additional information
construction of an isogenic deletion mutant strain DELTAcgh, the growth of the the cgh mutant is inhibited by addition of 5% or 10% bile acid, but is similar to the wild-type strain in absence of bile acids, the mutant enzyme does not confer resistance to bile acids to the organism
additional information
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construction of an isogenic deletion mutant strain DELTAcgh, the growth of the the cgh mutant is inhibited by addition of 5% or 10% bile acid, but is similar to the wild-type strain in absence of bile acids, the mutant enzyme does not confer resistance to bile acids to the organism
additional information
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construction of an isogenic deletion mutant strain DELTAcgh, the growth of the the cgh mutant is inhibited by addition of 5% or 10% bile acid, but is similar to the wild-type strain in absence of bile acids, the mutant enzyme does not confer resistance to bile acids to the organism
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additional information
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BSH is microencapsulated by foodgrade whey protein-gum arabic, comparison to living Lactobacillus plantarum WCFS1 in vitro, endogenously producing BSH, for its catalytic efficacy. BSH efficacy is better against pancreatin and low gastric pH initially, but its activity decreases further due to proteolytic degradation, whereas WCFS1 cells withstands such conditions. Bile salt deconjugation rates of microencapsulated BSH overproducing cells of strain 80 are 0.0049 mmol/g microcapsule per h towards glycoconjugates and 0.00079 mmol/g microcapsule per h towards tauroconjugates in the simulated gastro-intestine
additional information
generation of gene bsh deletion mutant Lactobacillus plantarum strains
additional information
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generation of gene bsh deletion mutant Lactobacillus plantarum strains
additional information
three twin-arginine signal peptides from twin-arginine translocation (Tat) pathway are synthesized, fused with bsh gene, inserted into expression vectors pET-20b(+) and pET-22b(+), and transformed into four different Escherichia coli hosts, respectively. Periplasmic export across the cytoplasmic membrane of bile salt hydrolase expressed in recombinant Escherichia coli by the twin-arginine signal peptides is achieved. Among the 24 recombinant bacteria obtained, Escherichia coli strain BL21(DE3) pLysS (pET-20b(+)-dmsA-bsh) shows the highest BSH activity in periplasmic fraction, which is further increased to 1.21 U/ml by orthogonal experimental design, overview. The presence of BSH in the periplasm is proven to be caused by the export rather than cell leakage. Export across the cytoplasmic membrane is initiated by the interaction of the signal peptides with the Tat export machinery, which in Escherichia coli is made up of the membrane protein TatABC
additional information
Lactiplantibacillus plantarum ATCC BAA-793
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generation of gene bsh deletion mutant Lactobacillus plantarum strains
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additional information
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three twin-arginine signal peptides from twin-arginine translocation (Tat) pathway are synthesized, fused with bsh gene, inserted into expression vectors pET-20b(+) and pET-22b(+), and transformed into four different Escherichia coli hosts, respectively. Periplasmic export across the cytoplasmic membrane of bile salt hydrolase expressed in recombinant Escherichia coli by the twin-arginine signal peptides is achieved. Among the 24 recombinant bacteria obtained, Escherichia coli strain BL21(DE3) pLysS (pET-20b(+)-dmsA-bsh) shows the highest BSH activity in periplasmic fraction, which is further increased to 1.21 U/ml by orthogonal experimental design, overview. The presence of BSH in the periplasm is proven to be caused by the export rather than cell leakage. Export across the cytoplasmic membrane is initiated by the interaction of the signal peptides with the Tat export machinery, which in Escherichia coli is made up of the membrane protein TatABC
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additional information
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knockout of gene bshA gene of strain NCFM decreases the ability to hydrolyze the chenodeoxycholic containing bile salts, e.g., taurochenodeoxycholic acid and glycochenodeoxycholic acid
additional information
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BSH from ATCC 4005 is immobilized in 0.5% gellan gum gel, method, overview
additional information
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BSH from strain ATCC 4005 is immobilized on 0.5% gellan gum gel for potential economic utilization
additional information
recombinant expression of BSH1 in Lactococcus lactis strain NZ9000 results in Lactococcus lactis strain NZ-bsh1 which shows a 2.8fold increased biomass compared to wild-type. The wild-type cytoplasm becomes sparse after exposure to porcine bile salt, and the thickness and smoothness of the cytomembranes are decreased. By contrast, there is no significant decline in the compactness of NZ-bsh1 or NZ-bsh2 cytoplasm under the same stress. Moreover, vesicle-like components emerge in the cytoplasm of cells expressing BSHs, and a similar component is formed around the cytomembrane's surface
additional information
recombinant expression of BSH1 in Lactococcus lactis strain NZ9000 results in Lactococcus lactis strain NZ-bsh1 which shows a 2.8fold increased biomass compared to wild-type. The wild-type cytoplasm becomes sparse after exposure to porcine bile salt, and the thickness and smoothness of the cytomembranes are decreased. By contrast, there is no significant decline in the compactness of NZ-bsh1 or NZ-bsh2 cytoplasm under the same stress. Moreover, vesicle-like components emerge in the cytoplasm of cells expressing BSHs, and a similar component is formed around the cytomembrane's surface
additional information
the pBSH plasmid bearing original bsh gene from Lactobacillus salivarius NRRL B-30514 is used as parent vector for site-directed mutagenesis. Construction of truncated mutant DELTA164-171, which is catalytically inactive
additional information
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the pBSH plasmid bearing original bsh gene from Lactobacillus salivarius NRRL B-30514 is used as parent vector for site-directed mutagenesis. Construction of truncated mutant DELTA164-171, which is catalytically inactive
additional information
Ligilactobacillus salivarius LMG14476
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recombinant expression of BSH1 in Lactococcus lactis strain NZ9000 results in Lactococcus lactis strain NZ-bsh1 which shows a 2.8fold increased biomass compared to wild-type. The wild-type cytoplasm becomes sparse after exposure to porcine bile salt, and the thickness and smoothness of the cytomembranes are decreased. By contrast, there is no significant decline in the compactness of NZ-bsh1 or NZ-bsh2 cytoplasm under the same stress. Moreover, vesicle-like components emerge in the cytoplasm of cells expressing BSHs, and a similar component is formed around the cytomembrane's surface
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additional information
Ligilactobacillus salivarius NRRL B-30514
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the pBSH plasmid bearing original bsh gene from Lactobacillus salivarius NRRL B-30514 is used as parent vector for site-directed mutagenesis. Construction of truncated mutant DELTA164-171, which is catalytically inactive
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additional information
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the enzyme is immobilized by micro-encapsulation. The microcapsules metabolize glyco- and tauroconjugated bile salts at rates of 0.0102 and 0.00185 mmol/g microcapsule per hour, respectively, and show better acid resistance. They exhibit an improved deconjugation with 49.4% of glycoconjugates per h in a simulated human gastrointestinal model and complete deconjugation after 4 h