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Expression, purification and crystallization of 2-oxo-hept-4-ene-1,7-dioate hydratase (HpcG) from Escherichia coli C
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Adachi, Tomoko, Izumi, Atsushi, Rea, Dean, Park, Sam-Yong, Tame, Jeremy R. H. and Roper, David I. (2006) Expression, purification and crystallization of 2-oxo-hept-4-ene-1,7-dioate hydratase (HpcG) from Escherichia coli C. Acta Crystallographica Section F-Structural Biology And Crystallization Communicati, 62 (Part 10). pp. 1010-1012. doi:10.1107/S1744309106035901 ISSN 1744-3091.
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Official URL: http://dx.doi.org/10.1107/S1744309106035901
Abstract
The gene encoding 2-oxo-hept-3-ene-1,7-dioic acid (OHED) hydratase (HpcG) was cloned into the high-expression plasmid pET26b and overexpressed in Escherichia coli BL21(DE3). The enzyme was purified in three steps to greater than 95% purity prior to crystallization. Crystals were obtained by the hanging-drop vapour-diffusion method at 277 K in a number of screening conditions. Crystals measuring up to 1.5 mm in their longest dimension were grown from solutions containing polyethylene glycol 20 000. The crystals belonged to space group P4(1)2(1)2 or P4(3)2(1)2, with unit-cell parameters a = 136, b = 136, c = 192 angstrom. A complete data set was collected to 2.1 angstrom from a single cryocooled crystal at 100 K using synchrotron radiation.
Item Type: | Journal Article | ||||
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Subjects: | Q Science > QD Chemistry Q Science > QH Natural history > QH301 Biology |
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Journal or Publication Title: | Acta Crystallographica Section F-Structural Biology And Crystallization Communicati | ||||
Publisher: | Blackwell Publishing | ||||
ISSN: | 1744-3091 | ||||
Official Date: | October 2006 | ||||
Dates: |
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Volume: | 62 | ||||
Number: | Part 10 | ||||
Number of Pages: | 3 | ||||
Page Range: | pp. 1010-1012 | ||||
DOI: | 10.1107/S1744309106035901 | ||||
Publication Status: | Published |
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