Abstract
Methylmalonyl-CoA epimerase (MCE) from the hyperthermophilic archaeon, Pyrococcus horikoshii, was expressed at high levels in Escherichia coli, purified, and partially characterized. The P. horikoshii MCE enzyme was a homodimer with an apparent molecular mass of 31,700 Da. The K m of the enzyme for methylmalonyl-CoA was 79 μM and the k cat was 240 s−1. The P. horikoshii enzyme was extremely heat-stable and withstood boiling for 60 min without detectable loss in activity.
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Acknowledgements
This work was supported by grant MCB-9876212 from the National Science Foundation, by grant GM59486 from the National Institutes of Health, and by the Florida Agricultural Experiment Station. This publication is Florida Agricultural Experiment Station Journal Series number R-09816. The experiments performed in this work comply with current laws of the United States of America.
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Bobik, T.A., Rasche, M.E. Purification and partial characterization of the Pyrococcus horikoshii methylmalonyl-CoA epimerase. Appl Microbiol Biotechnol 63, 682–685 (2004). https://doi.org/10.1007/s00253-003-1474-5
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DOI: https://doi.org/10.1007/s00253-003-1474-5