The purification and characterization of methylmalonyl CoA mutase from bovine brain
Methylmalonyl CoA mutase has been purified over 2300-fold from bovine brain using fractional precipitation, ion exchange resins, and gel filtration procedures. The crude extract had an equal mixture of mutase in the halo- and apoenzyme form. After the final purification step the ratio had changed to...
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| Format: | Others |
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BYU ScholarsArchive
1974
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| Online Access: | https://scholarsarchive.byu.edu/etd/8305 https://scholarsarchive.byu.edu/cgi/viewcontent.cgi?article=9305&context=etd |
| Summary: | Methylmalonyl CoA mutase has been purified over 2300-fold from bovine brain using fractional precipitation, ion exchange resins, and gel filtration procedures. The crude extract had an equal mixture of mutase in the halo- and apoenzyme form. After the final purification step the ratio had changed to 86% holo-enzyme and 14% apoenzyme. The mutase enzyme had a pH optimum of 7. 0 in Tris-HCI buffer. The Km values for L-methylmalonyl CoA and succinyl CoA were 7.7 x 10^-4 M and 1.8 x 10^-4 M respectively. The equiIibrium constant in the direction of succinyl CoA formation was 19. Inhibition with N-ethylmalei-mide was noncompetitive with a K_i of 2.4 x 10^-3 M. The activity level of mutase in the brain was found to be about 5% of that found in liver. |
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