Expression, purification and preliminary crystallographic analysis of 2,4-dihydroxy-hepta-2-ene-1,7-dioate aldolase (HpcH) from Escherichia coli C

Acta Crystallographica. Section F, Structural Biology and Crystallization Communications
Dean ReaD I Roper

Abstract

The gene encoding 2,4-dihydroxy-hepta-2-ene-1,7-dioate (HHED) aldolase (HpcH; EC 4.1.2) from Escherichia coli C was cloned into the high-expression plasmid pProEx-HTa and overexpressed in E. coli BL21 (DE3). The 28 kDa enzyme was purified using immobilized metal-affinity and size-exclusion chromatography prior to crystallization. Crystals were obtained by the hanging-drop vapour-diffusion method at 277 K from a number of screening conditions. Type I crystals grown in a solution containing 0.4 M ammonium dihydrogen phosphate belong to space group R32, with unit-cell parameters a = b = 128.92, c = 175.30 A. Type II crystals grown in a solution containing 0.5 M sodium chloride, 0.1 M sodium citrate pH 5.5 belong to space group I222, with unit-cell parameters a = 133.39, b = 155.39, c = 168.80 A. Complete data sets were collected to 1.6 and 2.0 A from type I and type II crystals, respectively, using synchrotron radiation.

References

Dec 1, 2001·Microbiology and Molecular Biology Reviews : MMBR·E DíazJ L García
Nov 28, 2002·Acta Crystallographica. Section D, Biological Crystallography·Anne WrightDavid Roper
Feb 3, 2005·Biotechnology and Bioengineering·Angela R Bielefeldt, Todd Cort

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Citations

Oct 3, 2006·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Tomoko AdachiDavid I Roper
Dec 25, 2012·Chemistry & Biology·Johnson CheungDavid E Heinrichs

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