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Cloning, purification, and crystallization of Escherichia coli cystathionine β‐lyase
Author(s) -
Laber Bernd,
Clausen Tim,
Huber Robert,
Messerschmidt Albrecht,
Egner Ursula,
Müller-Fahrnow Anke,
Pohlenz Hans-Dieter
Publication year - 1996
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(95)01499-3
Subject(s) - escherichia coli , cystathionine beta synthase , ammonium sulfate , dimer , chemistry , cystathionine gamma lyase , crystallization , cloning (programming) , lyase , biochemistry , enzyme , chromatography , gene , cysteine , organic chemistry , computer science , programming language
The metC gene coding for cystathionine β‐lyase of Escherichia coli has been cloned and used to construct an overproducing E. coli strain. An efficient purification scheme has been developed and the purified enzyme has been crystallized by the hanging drop vapour diffusion method using either ammonium sulfate or polyethyleneglycol 400 as precipitating agent. The crystals belong to the orthorombic space group C222 1 . Their unit cell parameters are . Consideration of the possible values of V M accounts for the presence of one dimer per asymmetric unit. The crystals are suitable for X‐ray analysis and a complete native date set to 1.83 Å resolution has been collected using synchrotron radiation.