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2.5.1.47: cysteine synthase

This is an abbreviated version!
For detailed information about cysteine synthase, go to the full flat file.

Word Map on EC 2.5.1.47

Reaction

O-acetyl-L-serine
+
hydrogen sulfide
=
L-cysteine
 +
acetate

Synonyms

(OAS-TL)-like protein, acetylserine sulfhydrylase, At-OASS, ATCC 23270 protein, BAbS19_I09950, beta-cyano-alanine synthase, beta-cyano-L-alanine synthase, CS-A, CS-B, CS1, CS26, CS3, CSase, CSase A, CSase B, CSaseLP, CSC, Cys synthase complex, cysK, CysK1, CysK2, CYSL, CysL-1, cysl-2, CysM, Cysteine synthase, cysteine synthase 1, cysteine synthase A, cysteine synthase B, cysteine synthase, chloroplastic/chromoplastic, cysteine synthase, mitochondrial, cysteine synthetase, DcsD, EC 4.2.99.8, EhCS, EhCS1, EhCS3, EhOASS, Fn1220, GmOAS-TL1, GmOAS-TL2, GmOAS-TL3, GmOAS-TL4, GmOAS-TL6, GmOAS-TL7, GmOASTL4, GsOAS-TL1, GYY4137, HiOASS, HiOASS-A, lanthionine synthase, LbrCS, MCSA1, Nt-OAS-TL, O -acetylserine (thiol)-lyase, O-acetyl-L-serine (thiol) lyase, O-acetyl-L-serine acetate-lyase (adding hydrogen sulfide), O-acetyl-L-serine sulfhydrylase, O-acetyl-L-serine sulfhydrylase B, O-acetyl-L-serine sulfohydrolase, O-acetyl-L-serine(thiol)lyase, O-acetyl-serine (thiol) lyase, O-acetylserine (thiol) lyase, O-acetylserine (Thiol)-lyase, O-acetylserine (thiol)-lyase A, O-acetylserine (thiol)-lyase B, O-acetylserine (thiol)lyase, O-acetylserine sulfhydrylase, O-acetylserine sulfhydrylase A, O-acetylserine sulfhydrylase B, O-acetylserine sulfhydrylase isoenzyme B, O-acetylserine sulfhydrylase-A, O-acetylserine sulfhydrylase-B, O-acetylserine thiol lyase, O-acetylserine(thiol)lyase, O-acetylserine-(thiol)lyase, O-acetylserine-O-acetylhomoserine sulfhydro-lyase, O3-acetyl-L-serine:hydrogen-sulfide 2-amino-2-carboxyethyltransferase, OAS Shase, OAS thiol-lyases, OAS-TL, OAS-TL A, OAS-TL B, OAS-TL C, OASA1, OASB, OASS, OASS-A, OASS-B, OASTL, OASTL-A, OASTL-A1, old3-1, OPSS, Os01g0290600, Os03g53650, protein ONSET OF LEAF DEATH 3, RCS3, S-sulfocysteine synthase, SSC synthase, StOASTL A, StOASTL B, sulfhydrylase/cysteine synthase, synthase, cysteine, TvCS1

ECTree

     2 Transferases
         2.5 Transferring alkyl or aryl groups, other than methyl groups
             2.5.1 Transferring alkyl or aryl groups, other than methyl groups (only sub-subclass identified to date)
                2.5.1.47 cysteine synthase

Purification

Purification on EC 2.5.1.47 - cysteine synthase

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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
cell harvesting are resuspended in 50 mM Tris buffer, pH 8.0, containing 10 mM EDTA, 15 mM beta-mercaptoethanol, 0.05 mM N-p-tosyl-L-lysine chloromethylketone, leupeptin, aprotinin, pepstatin, lysozyme and pyridoxal 5'-phosphate, centrifuged, supernatant loaded on a DE52 anion exchange column, washed with 50 mM Tris buffer, pH 8.0, with 10 mM EDTA, and 15 mM beta-mercaptoethanol, elution with NaCl gradient in buffer, active fractions are pooled, concentrated and applied to a GSTrap 4B column, washed with PBS buffer, pH 7.4, containing 140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, and 1.8 mM KH2PO4, elution with 50 mM Tris, pH 8.0, with 20 mM reduced glutathione and thrombin, dialyzed with 15 mM potassium phosphate, pH 7.2, applied on a DE 52 column, separation of enzyme from GST-tag and thrombin with a gradient of 15-300 mM potassium phosphate buffer, pH 7.2
-
cells are harvested by centrifugation, washed with sterile water, again harvested by centrifugation, suspended in 20 mM potassium phosphate buffer, pH 7.4, containing 0.5 M NaCl, lysed with lysozyme at room temperature, one-step affinity chromatography with nickel metal-affinity resin columns, dialyzed against 20 mM potassium phosphate buffer, pH 7.5, 5% glycerol, and 5 mM 2-mercaptoethanol
-
ethanol precipitation, ammonium sulfate precipitation, preparative electrophoresis, phenyl sepharose
His-tagged recombinant protein
isoenzyme A and B
-
Ni-NTA affinity and Superdex 200 pg gel filtration chromatography
purified using nickel-affinity and gel filtration. Thrombin digestion is used to remove the His tag from each protein
Q-Sepharose and phenyl Sepharose column, active fractions are pooled and concentrated
-
recombinant chloroplast isozyme OASTL 29fold from Chlorella sorokiniana S-sufficient and S-starved cells by affinity binding to Arabidopsis thaliana serine acetyltransferase SAT5 fused to a metal resin via its His-tag protein, elution by by O-acetylserine, to homogeneity. Successful application of SAT/OASTL interaction for purification
-
recombinant enzyme
-
recombinant His-tagged enzyme from Escherichia coli strain BL21 (DE3) pLYsS by nickel affinity chhromatography
recombinant His-tagged enzyme from Escherichia coli strain BL21 (DE3) pLysS by nickel affinity chromatography and gel filtration
-
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography to homogeneity and over 95% purity
recombinant N-terminal His-tagged enzyme, cleavage and removal of the tag
recombinant N-terminally His6-tagged isozyme CysK1 from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography and ultrafiltration
recombinant N-terminally His6-tagged isozyme CysK2 from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography and ultrafiltration
recombinant protein
recombinant protein of isoforms A,B,C
-
Source 30Q column chromatography and Superdex 75 gel filtration
the apoenzyme of OASS-B is prepared using hydroxylamine as the resolving reagent. Apoenzyme is reconstituted to holoenzyme by addition of pyridoxal 5'-phosphate
-
the soluble protein is purified by one-step affinity chromatography to apparent homogeneity
-
using a GSTrap 4 B column and anion-exchange chromatography
-
using Glutathione Sepharose 4B column
using Ni-NTA chromatography
using Ni-NTA chromatography and gel filtration