Abstract:
:Steady-state kinetic studies of the bovine carbonic anhydrase B-catalyzed hydration of CO2, dehydration of HCO3-, and hydrolysis of p-nitrophenylacetate were made in glycerol/water solvents of increased viscosity in order that the effect of diffusion-control on the substrate association reactions could be determined. The minimum association rate constants (kmin = V/(Km[E0])) were obtained at low substrate concentrations. The esterase activity did not depend upon the solvent viscosity. However, both the CO2 hydration and HCO3- dehydration reactions depended upon the solvent viscosity consistent with partial diffusion control. Thus both chemical activation and diffusion control processes contribute to the observed kmin. In low-viscosity aqueous solutions both hydration and dehydration are largely controlled by chemical activation. However, at higher viscosities, equal to that found in the interior of the erythrocyte, both reactions are largely diffusion controlled. This result can be interpreted to mean that carbonic anhydrase is a highly evolved enzyme that has approached its maximum efficiency. The extent of diffusion control observed rules out H2CO3 as a significant reactant with the enzyme. Several models that yield minimum steric requirements for access of substrate to the active site are examined. Minimum steric constraints are less for the smaller CO2. The slower esterase reaction is not influenced by diffusion.
journal_name
Arch Biochem Biophysjournal_title
Archives of biochemistry and biophysicsauthors
Hasinoff BBdoi
10.1016/0003-9861(84)90494-6subject
Has Abstractpub_date
1984-09-01 00:00:00pages
676-81issue
2eissn
0003-9861issn
1096-0384pii
0003-9861(84)90494-6journal_volume
233pub_type
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journal_title:Archives of biochemistry and biophysics
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pub_type: 杂志文章
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更新日期:2012-01-15 00:00:00
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pub_type: 杂志文章
doi:10.1016/0003-9861(92)90421-r
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pub_type: 杂志文章
doi:10.1016/j.abb.2011.07.013
更新日期:2011-10-01 00:00:00
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pub_type: 杂志文章
doi:10.1006/abbi.2000.2004
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pub_type: 杂志文章
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journal_title:Archives of biochemistry and biophysics
pub_type: 杂志文章
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更新日期:1989-06-01 00:00:00
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pub_type: 杂志文章
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更新日期:1985-11-15 00:00:00
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journal_title:Archives of biochemistry and biophysics
pub_type: 杂志文章
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更新日期:2018-05-01 00:00:00